Patent Publication Number: US-2020282074-A1

Title: Nucleic acid-polypeptide compositions and methods of inducing exon skipping

Description:
CROSS-REFERENCE 
     This application claims the benefit of U.S. Provisional Application No. 62/561,939, filed Sep. 22, 2017, and U.S. Provisional Application No. 62/696,766, filed Jul. 11, 2018, each of which is incorporated herein by reference in its entirety. 
    
    
     BACKGROUND OF THE DISCLOSURE 
     Modulation of RNA function is a developing area of therapeutic interest. Drugs that affect mRNA stability like antisense oligonucleotides and short interfering RNAs are one way to modulate RNA function. Another group of oligonucleotides can modulate RNA function by altering the processing of pre-mRNA to include or exclude specific regions of pre-mRNAs from the ultimate gene product: the encoded protein. As such, oligonucleotide therapeutics represent a means of modulating protein expression in disease states and as such have utility as therapeutics. 
     SUMMARY OF THE DISCLOSURE 
     Disclosed herein, in certain embodiments, are molecules and pharmaceutical compositions for modulating RNA processing. In some embodiments, also disclosed herein are molecules and pharmaceutical compositions for the treatment of a muscular dystrophy. 
     Disclosed herein, in certain embodiments, are methods of treating a disease or disorder caused by an incorrectly spliced mRNA transcript in a subject in need thereof, the method comprising: administering to the subject a polynucleic acid molecule conjugate; wherein the polynucleic acid molecule conjugate is conjugated to a cell targeting binding moiety; wherein the polynucleotide optionally comprises at least one 2′ modified nucleotide, at least one modified internucleotide linkage, or at least one inverted abasic moiety; wherein the polynucleic acid molecule conjugate induces insertion, deletion, duplication, or alteration in the incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion in the incorrectly spliced mRNA transcript to generate a fully processed mRNA transcript; and wherein the fully processed mRNA transcript encodes a functional protein, thereby treating the disease or disorder in the subject. In some embodiments, the disease or disorder is further characterized by one or more mutations in the mRNA. In some embodiments, the disease or disorder comprises a neuromuscular disease, a genetic disease, cancer, a hereditary disease, or a cardiovascular disease. In some embodiments, the disease or disorder is muscular dystrophy. In some embodiments, the disease or disorder is Duchenne muscular dystrophy. In some embodiments, the exon skipping is of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some embodiments, the exon skipping is of exon 23 of the DMD gene. In some embodiments, the polynucleic acid molecule conjugate is of Formula (I): 
       A-X—B   Formula I
 
     wherein, 
     A is a binding moiety; 
     B is a polynucleotide; and 
     X is a bond or first linker. 
     In some embodiments, the polynucleic acid molecule conjugate is of Formula (II): 
       A-X—B—Y—C   Formula II
 
     wherein, 
     A is a binding moiety; 
     B is a polynucleotide; 
     C is a polymer; 
     X is a bond or first linker; and 
     Y is a bond or second linker. 
     In some embodiments, the polynucleic acid molecule conjugate is of Formula (III): 
       A-X—C—Y—B   Formula III
 
     wherein, 
     A is a binding moiety; 
     B is a polynucleotide; 
     C is a polymer; 
     X is a bond or first linker; and 
     Y is a bond or second linker. 
     In some embodiments, the at least one 2′ modified nucleotide comprises a morpholino, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified nucleotide. In some embodiments, the at least one 2′ modified nucleotide comprises locked nucleic acid (LNA), ethylene nucleic acid (ENA), or a peptide nucleic acid (PNA). In some embodiments, the at least one 2′ modified nucleotide comprises a morpholino. In some embodiments, the at least one inverted basic moiety is at least one terminus. In some embodiments, the at least one modified internucleotide linkage comprises a phosphorothioate linkage or a phosphorodithioate linkage. In some embodiments, the polynucleic acid molecule is at least from about 10 to about 30 nucleotides in length. In some embodiments, the polynucleic acid molecule is at least one of: from about 15 to about 30, from about 18 to about 25, from about 18 to about 24, from about 19 to about 23, or from about 20 to about 22 nucleotides in length. In some embodiments, the polynucleic acid molecule is at least about 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in length. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 5% to about 100% modification, from about 10% to about 100% modification, from about 20% to about 100% modification, from about 30% to about 100% modification, from about 40% to about 100% modification, from about 50% to about 100% modification, from about 60% to about 100% modification, from about 70% to about 100% modification, from about 80% to about 100% modification, and from about 90% to about 100% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 90% modification, from about 20% to about 90% modification, from about 30% to about 90% modification, from about 40% to about 90% modification, from about 50% to about 90% modification, from about 60% to about 90% modification, from about 70% to about 90% modification, and from about 80% to about 100% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 80% modification, from about 20% to about 80% modification, from about 30% to about 80% modification, from about 40% to about 80% modification, from about 50% to about 80% modification, from about 60% to about 80% modification, and from about 70% to about 80% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 70% modification, from about 20% to about 70% modification, from about 30% to about 70% modification, from about 40% to about 70% modification, from about 50% to about 70% modification, and from about 60% to about 70% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 60% modification, from about 20% to about 60% modification, from about 30% to about 60% modification, from about 40% to about 60% modification, and from about 50% to about 60% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 50% modification, from about 20% to about 50% modification, from about 30% to about 50% modification, and from about 40% to about 50% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 40% modification, from about 20% to about 40% modification, and from about 30% to about 40% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 30% modification, and from about 20% to about 30% modification. In some embodiments, the polynucleic acid molecule comprises from about 10% to about 20% modification. In some embodiments, the polynucleic acid molecule comprises from about 15% to about 90%, from about 20% to about 80%, from about 30% to about 70%, or from about 40% to about 60% modifications. In some embodiments, the polynucleic acid molecule comprises at least about 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% modification. In some embodiments, the polynucleic acid molecule comprises at least about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22 or more modifications. In some embodiments, the polynucleic acid molecule comprises at least about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22 or more modified nucleotides. In some embodiments, the polynucleic acid molecule comprises a single strand. In some embodiments, the polynucleic acid molecule comprises two or more strands. In some embodiments, the polynucleic acid molecule comprises a first polynucleotide and a second polynucleotide hybridized to the first polynucleotide to form a double-stranded polynucleic acid molecule. In some embodiments, the second polynucleotide comprises at least one modification. In some embodiments, the first polynucleotide and the second polynucleotide are RNA molecules. In some embodiments, the first polynucleotide and the second polynucleotide are siRNA molecules. In some embodiments, X and Y are independently a bond, a degradable linker, a non-degradable linker, a cleavable linker, or a non-polymeric linker group. In some embodiments, X is a bond. In some embodiments, X is a C 1 -C 6  alkyl group. In some embodiments, Y is a C 1 -C 6  alkyl group. In some embodiments, X is a homobifuctional linker or a heterobifunctional linker, optionally conjugated to a C 1 -C 6  alkyl group. In some embodiments, Y is a homobifuctional linker or a heterobifunctional linker. In some embodiments, the binding moiety is an antibody or binding fragment thereof. In some embodiments, the antibody or binding fragment thereof comprises a humanized antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab′, divalent Fab2, single-chain variable fragment (scFv), diabody, minibody, nanobody, single-domain antibody (sdAb), or camelid antibody or binding fragment thereof. In some embodiments, C is polyethylene glycol. In some embodiments, C has a molecular weight of about 5000 Da. In some embodiments, A-X is conjugated to the 5′ end of B and Y—C is conjugated to the 3′ end of B. In some embodiments, Y—C is conjugated to the 5′ end of B and A-X is conjugated to the 3′ end of B. In some embodiments, A-X, Y—C or a combination thereof is conjugated to an internucleotide linkage group. In some embodiments, methods further comprise D. In some embodiments, D is conjugated to C or to A. In some embodiments, D is conjugated to the molecule conjugate of Formula (II) according to Formula (IV): 
       (A-X—B—Y—C c )-L-D   Formula IV
         wherein,   A is a binding moiety;   B is a polynucleotide;   C is a polymer;   X is a bond or first linker;   Y is a bond or second linker;   L is a bond or third linker;   D is an endosomolytic moiety; and   c is an integer between 0 and 1; and   wherein the polynucleotide comprises at least one 2′ modified nucleotide, at least one modified internucleotide linkage, or an inverted abasic moiety; and D is conjugated anywhere on A, B, or C.
 
In some embodiments, D is INF7 or melittin. In some embodiments, L is a C 1 -C 6  alkyl group. In some embodiments, L is a homobifuctional linker or a heterobifunctional linker. In some embodiments, methods further comprise at least a second binding moiety A. In some embodiments, the at least second binding moiety A is conjugated to A, to B, or to C.
       

     Disclosed herein, in some embodiments, are methods of inducing an insertion, deletion, duplication, or alteration in the incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion in the incorrectly spliced mRNA transcript, the method comprising: contacting a target cell with a polynucleic acid molecule conjugate, wherein the polynucleotide comprises at least one 2′ modified nucleotide, at least one modified internucleotide linkage, or at least one inverted abasic moiety; hybridizing the polynucleic acid molecule conjugate to the incorrectly spliced mRNA transcript within the target cell to induce an insertion, deletion, duplication, or alteration in the incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion, wherein the incorrectly spliced mRNA transcript is capable of encoding a functional form of a protein; and translating the functional form of a protein from a fully processed mRNA transcript of the previous step. In some embodiments, the target cell is a target cell of a subject. In some embodiments, the incorrectly spliced mRNA transcript further induces a disease or disorder. In some embodiments, the disease or disorder is further characterized by one or more mutations in the mRNA. In some embodiments, the disease or disorder comprises a neuromuscular disease, a genetic disease, cancer, a hereditary disease, or a cardiovascular disease. In some embodiments, the disease or disorder is muscular dystrophy. In some embodiments, the disease or disorder is Duchenne muscular dystrophy. In some embodiments, the exon skipping is of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some embodiments, the exon skipping is of exon 23 of the DMD gene. In some embodiments, the polynucleic acid molecule conjugate is of Formula (I): 
       A-X—B   Formula I
 
     wherein, 
     A is a binding moiety; 
     B is a polynucleotide; and 
     X is a bond or first linker. 
     In some embodiments, the polynucleic acid molecule conjugate is of Formula (II): 
       A-X—B—Y—C   Formula II
 
     wherein, 
     A is a binding moiety; 
     B is a polynucleotide; 
     C is a polymer; 
     X is a bond or first linker; and 
     Y is a bond or second linker. 
     In some embodiments, the polynucleic acid molecule conjugate is of Formula (III): 
       A-X—C—Y—B   Formula III
 
     wherein, 
     A is a binding moiety; 
     B is a polynucleotide; 
     C is a polymer; 
     X is a bond or first linker; and 
     Y is a bond or second linker. 
     In some embodiments, the at least one 2′ modified nucleotide comprises a morpholino, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified nucleotide. In some embodiments, the at least one 2′ modified nucleotide comprises locked nucleic acid (LNA), ethylene nucleic acid (ENA), peptide nucleic acid (PNA). In some embodiments, the at least one 2′ modified nucleotide comprises a morpholino. In some embodiments, the at least one inverted basic moiety is at least one terminus. In some embodiments, the at least one modified internucleotide linkage comprises a phosphorothioate linkage or a phosphorodithioate linkage. In some embodiments, the polynucleic acid molecule is at least from about 10 to about 30 nucleotides in length. In some embodiments, the polynucleic acid molecule is at least one of: from about 15 to about 30, from about 18 to about 25, from about 18 to about 24, from about 19 to about 23, or from about 20 to about 22 nucleotides in length. In some embodiments, the polynucleic acid molecule is at least about 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in length. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 5% to about 100% modification, from about 10% to about 100% modification, from about 20% to about 100% modification, from about 30% to about 100% modification, from about 40% to about 100% modification, from about 50% to about 100% modification, from about 60% to about 100% modification, from about 70% to about 100% modification, from about 80% to about 100% modification, and from about 90% to about 100% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 90% modification, from about 20% to about 90% modification, from about 30% to about 90% modification, from about 40% to about 90% modification, from about 50% to about 90% modification, from about 60% to about 90% modification, from about 70% to about 90% modification, and from about 80% to about 100% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 80% modification, from about 20% to about 80% modification, from about 30% to about 80% modification, from about 40% to about 80% modification, from about 50% to about 80% modification, from about 60% to about 80% modification, and from about 70% to about 80% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 70% modification, from about 20% to about 70% modification, from about 30% to about 70% modification, from about 40% to about 70% modification, from about 50% to about 70% modification, and from about 60% to about 70% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 60% modification, from about 20% to about 60% modification, from about 30% to about 60% modification, from about 40% to about 60% modification, and from about 50% to about 60% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 50% modification, from about 20% to about 50% modification, from about 30% to about 50% modification, and from about 40% to about 50% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 40% modification, from about 20% to about 40% modification, and from about 30% to about 40% modification. In some embodiments, the polynucleic acid molecule comprises at least one of: from about 10% to about 30% modification, and from about 20% to about 30% modification. In some embodiments, the polynucleic acid molecule comprises from about 10% to about 20% modification. In some embodiments, the polynucleic acid molecule comprises from about 15% to about 90%, from about 20% to about 80%, from about 30% to about 70%, or from about 40% to about 60% modifications. In some embodiments, the polynucleic acid molecule comprises at least about 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% modification. In some embodiments, the polynucleic acid molecule comprises at least about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22 or more modifications. In some embodiments, the polynucleic acid molecule comprises at least about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22 or more modified nucleotides. In some embodiments, the polynucleic acid molecule comprises a single strand. In some embodiments, the polynucleic acid molecule comprises two or more strands. In some embodiments, the polynucleic acid molecule comprises a first polynucleotide and a second polynucleotide hybridized to the first polynucleotide to form a double-stranded polynucleic acid molecule. In some embodiments, the second polynucleotide comprises at least one modification. In some embodiments, the first polynucleotide and the second polynucleotide are RNA molecules. In some embodiments, the first polynucleotide and the second polynucleotide are siRNA molecules. In some embodiments, X and Y are independently a bond, a degradable linker, a non-degradable linker, a cleavable linker, or a non-polymeric linker group. In some embodiments, X is a bond. In some embodiments, X is a C 1 -C 6  alkyl group. In some embodiments, Y is a C 1 -C 6  alkyl group. In some embodiments, X is a homobifuctional linker or a heterobifunctional linker, optionally conjugated to a C 1 -C 6  alkyl group. In some embodiments, Y is a homobifuctional linker or a heterobifunctional linker. In some embodiments, the binding moiety is an antibody or binding fragment thereof. In some embodiments, the antibody or binding fragment thereof comprises a humanized antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab′, divalent Fab2, single-chain variable fragment (scFv), diabody, minibody, nanobody, single-domain antibody (sdAb), or camelid antibody or binding fragment thereof. In some embodiments, C is polyethylene glycol. In some embodiments, C has a molecular weight of about 5000 Da. In some embodiments, A-X is conjugated to the 5′ end of B and Y—C is conjugated to the 3′ end of B. In some embodiments, Y—C is conjugated to the 5′ end of B and A-X is conjugated to the 3′ end of B. In some embodiments, A-X, Y—C or a combination thereof is conjugated to an internucleotide linkage group. In some embodiments, methods further comprise D. In some embodiments, D is conjugated to C or to A. In some embodiments, D is conjugated to the molecule conjugate of Formula (II) according to Formula (IV): 
       (A-X—B—Y—C c )-L-D   Formula IV
         wherein,   A is a binding moiety;   B is a polynucleotide;   C is a polymer;   X is a bond or first linker;   Y is a bond or second linker;   L is a bond or third linker;   D is an endosomolytic moiety; and   c is an integer between 0 and 1; and   wherein the polynucleotide comprises at least one 2′ modified nucleotide, at least one modified internucleotide linkage, or an inverted abasic moiety; and D is conjugated anywhere on A, B, or C.       

     In some embodiments, D is INF7 or melittin. In some embodiments, L is a C 1 -C 6  alkyl group. In some embodiments, L is a homobifuctional linker or a heterobifunctional linker. In some embodiments, methods further comprise at least a second binding moiety A. In some embodiments, the at least second binding moiety A is conjugated to A, to B, or to C. In some embodiments, the method is an in vivo method. In some embodiments, the method is an in vitro method. In some embodiments, the subject is a human. 
     Disclosed herein, in certain embodiments, are pharmaceutical compositions comprising: a molecule obtained by any one of the methods disclosed herein and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is formulated as a nanoparticle formulation. In some embodiments, the pharmaceutical composition is formulated for parenteral, oral, intranasal, buccal, rectal, or transdermal administration. 
     Disclosed herein, in certain embodiments, are compositions comprising a polynucleic acid molecule conjugate, wherein the polynucleic acid molecule conjugate comprises a polynucleotide comprising a sequence having at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 45-963. Disclosed herein, in certain embodiments, are compositions comprising a polynucleic acid molecule conjugate, wherein the polynucleic acid molecule conjugate comprises a polynucleotide comprising a sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 45-963. In certain embodiments, the polynucleic acid molecule conjugate is of Formula (I): 
       A-X—B   Formula I
 
     wherein, 
     A is a binding moiety; 
     B is the polynucleotide; and 
     X is a bond or first linker. 
     In certain embodiments, the polynucleic acid molecule conjugate is of Formula (II): 
       A-X—B—Y—C   Formula II
 
     wherein, 
     A is a binding moiety; 
     B is the polynucleotide; 
     C is a polymer; 
     X is a bond or first linker; and 
     Y is a bond or second linker. 
     In certain embodiments, the polynucleic acid molecule conjugate is of Formula (III): 
       A-X—C—Y—B   Formula III
 
     wherein, 
     A is a binding moiety; 
     B is the polynucleotide; 
     C is a polymer; 
     X is a bond or first linker; and 
     Y is a bond or second linker. 
     In certain embodiments, the at least one 2′ modified nucleotide comprises a morpholino, 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified nucleotide. In certain embodiments, the at least one 2′ modified nucleotide comprises a morpholino. 
     Disclosed herein, in certain embodiments, is a polynucleic acid conjugate comprising a target cell binding moiety binding to at least one polynucleic acid molecule that hybridizes to a target region of a pre-mRNA transcript of DMD gene, wherein the at least one polynucleic acid molecule induces splicing out of an exon from a pre-mRNA transcript to generate a mRNA transcript that encodes a functional dystrophin protein. In some embodiments, the functional dystrophin protein is a truncated form of the dystrophin protein. In some embodiments, the target region is at an exon-intron junction, wherein the exon is the exon that is to be spliced out. In some embodiments, the exon is exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55. In some embodiments, the exon-intron junction is located at the 5′ of the exon that is to be spliced out. In some embodiments, the target region is an intronic region upstream of the exon-intron junction. In some embodiments, the target region is about 500, 450, 400, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 nucleotides upstream of the exon-intron junction. In some embodiments, the exon-intron junction is located at the 3′ of the exon that is to be spliced out. In some embodiments, the target region is an intronic region downstream of the exon-intron junction. In some embodiments, the target region is about 500, 450, 400, 350, 300, 250, 200, 150, 100, 90, 80, 70, 60, 50, 40, 30, 20, or 10 nucleotides downstream of the exon-intron junction. In some embodiments, the target cell binding moiety binds to two or more, three or more, four or more, five or more, six or more, or eight or more polynucleic acid molecules. In some embodiments, the polynucleic acid molecule is from about 10 to about 50 nucleotides in length. In some embodiments, the polynucleic acid molecule comprises about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity to a sequence selected from SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule further comprises 1, 2, 3, or 4 mismatches. In some embodiments, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1056-1094, 1147-1162, or 1173-1211. In some embodiments, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1173-1211. In some embodiments, the binding moiety comprises an antibody. In some embodiments, the antibody comprises an anti-transferrin antibody. In some embodiments, the binding moiety comprises a plasma protein. In some embodiments, the polynucleic acid conjugate comprises A-(X 1 —B) n ; Formula (V), wherein, A comprises the binding moiety; B consists of the polynucleic acid molecule; X 1  consists of a bond or first non-polymeric linker; and n is an averaged value selected from 1-12. In some embodiments, the polynucleic acid molecule comprises a passenger strand and a guide strand. In some embodiments, the guide strand comprises at least one modified internucleotide linkage, at least one inverted abasic moiety, at least one 5′-vinylphosphonate modified non-natural nucleotide, or a combination thereof. In some embodiments, the guide strand comprises about 2, 3, 4, 5, 6, 7, 8, or 9 phosphorothioate-modified non-natural nucleotides. In some embodiments, the guide strand comprises 1 phosphorothioate-modified non-natural nucleotide. In some embodiments, the phosphorothioate modified non-natural nucleotide is located at an internucleotide linkage of the polynucleotide. In some embodiments, the at least one 5′-vinylphosphonate modified non-natural nucleotide is located about 1, 2, 3, 4, or 5 bases away from the 5′ terminus of the guide strand. In some embodiments, the at least one 5′-vinylphosphonate modified non-natural nucleotide is further modified at the 2′-position. In some embodiments, the 2′-modification is selected from 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified nucleotide. In some embodiments, the passenger strand comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorodiamidate morpholino oligomer-modified non-natural nucleotides. In some embodiments, the passenger strand comprises 100% phosphorodiamidate morpholino oligomer-modified non-natural nucleotides. In some embodiments, the passenger strand is shorter in length than the guide strand, thereby generating a 5′ overhang, a 3′ overhang, or a combination thereof. In some embodiments, the passenger strand is equal in length to the guide strand, thereby generating a blunt end at each terminus of the polynucleic acid molecule. In some embodiments, the polynucleic acid molecule is a phosphorodiamidate morpholino oligomer/RNA hetero-duplex. In some embodiments, the passenger strand comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more peptide nucleic acid-modified non-natural nucleotides. In some embodiments, the passenger strand comprises 100% peptide nucleic acid-modified non-natural nucleotides. In some embodiments, the passenger strand is shorter in length than the guide strand, thereby generating a 5′ overhang, a 3′ overhang, or a combination thereof. In some embodiments, the passenger strand is equal in length to the guide strand, thereby generating a blunt end at each terminus of the polynucleic acid molecule. In some embodiments, the polynucleic acid molecule is a peptide nucleic acid/RNA hetero-duplex. In some embodiments, the passenger strand is conjugated to A-X 1 . In some embodiments, A-X 1  is conjugated to the 5′ end of the passenger strand. In some embodiments, A-X 1  is conjugated to the 3′ end of the passenger strand. In some embodiments, X 1  is a bond. In some embodiments, X 1  is a C 1 -C 6  alkyl group. In some embodiments, X 1  is a homobifuctional linker or a heterobifunctional linker, optionally conjugated to a C 1 -C 6  alkyl group. In some embodiments, the polynucleic acid conjugate further comprises C. In some embodiments, C is polyethylene glycol. In some embodiments, C is directly conjugated to B via X 2 . In some embodiments, X 2  consists of a bond or second non-polymeric linker. In some embodiments, X 2  is a bond. In some embodiments, X 2  is a C 1 -C 6  alkyl group. In some embodiments, X 2  is a homobifuctional linker or a heterobifunctional linker, optionally conjugated to a C 1 -C 6  alkyl group. In some embodiments, the passenger strand is conjugated to A-X 1  and X 2 —C. In some embodiments, A-X 1  is conjugated to the 5′ end of the passenger strand and X 2 —C is conjugated to the 3′ end of the passenger strand. In some embodiments, X 2 —C is conjugated to the 5′ end of the passenger strand and A-X 1  is conjugated to the 3′ end of the passenger strand. In some embodiments, the polynucleic acid conjugate comprises: A-X 1 —(B—X 2 —C) n ; Formula (VI), wherein, A comprises the binding moiety; B consists of the polynucleic acid molecule; C consists of a polymer; X 1  consists a bond or first non-polymeric linker; X 2  consists of a bond or second non-polymeric linker; and n is an averaged value selected from 1-12. In some embodiments, the polynucleic acid conjugate further comprises D. In some embodiments, D is an endosomolytic moiety. 
     Disclosed herein, in certain embodiments, is a polynucleic acid molecule comprising at least 23 contiguous bases of a base sequence selected from SEQ ID NOs: 1056-1058 or 1087-1089, wherein the polynucleic acid molecule comprises no more than 50 nucleotides in length. 
     Disclosed herein, in certain embodiments, is a polynucleic acid molecule comprising SEQ ID NOs: 1056-1058, wherein the polynucleic acid molecule comprises no more than 50 nucleotides in length. 
     Disclosed herein, in certain embodiments, is a polynucleic acid molecule comprising SEQ ID NOs: 1087-1089, wherein the polynucleic acid molecule comprises no more than 50 nucleotides in length. 
     Disclosed herein, in certain embodiments, is a pharmaceutical composition, comprising: a polynucleic acid conjugate described herein or a polynucleic acid molecule described herein; and a pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is formulated for systemic delivery. In some embodiments, the pharmaceutical composition is formulated for parenteral administration. 
     Disclosed herein, in certain embodiments, is a method of treating a disease or condition characterized with a defective mRNA in a subject in need thereof, comprising: administering to the subject a polynucleic acid conjugate described herein or a polynucleic acid molecule described herein to induce skipping of an exon that leads to the defective mRNA to generate a processed mRNA encoding a functional protein, thereby treating the disease or condition in the subject. In some embodiments, the disease or condition is a neuromuscular disease, a genetic disease, cancer, a hereditary disease, or a cardiovascular disease. In some embodiments, the neuromuscular disease is a muscular dystrophy. In some embodiments, the muscular dystrophy is Duchenne muscular dystrophy, Becker muscular dystrophy, facioscapulohumeral muscular dystrophy, congenital muscular dystrophy, or myotonic dystrophy. In some embodiments, the subject is a human. 
     Disclosed herein, in certain embodiments, is a method of treating a muscular dystrophy in a subject in need thereof, comprising: administering to the subject a polynucleic acid conjugate described herein or a polynucleic acid molecule described herein, thereby treating the muscular dystrophy in the subject. In some embodiments, the muscular dystrophy is Duchenne muscular dystrophy. In some embodiments, the subject is a human. 
     Disclosed herein, in certain embodiments, is a kit comprising a polynucleic acid conjugate described herein or a polynucleic acid molecule described herein. 
     Disclosed herein, in certain embodiments, are kits comprising a molecule obtained by any one of the methods disclosed herein. 
    
    
     
       DESCRIPTION OF THE DRAWINGS 
         FIG. 1  depicts a phosphorodiamidate morpholino oligomer (PMO) sequence with end nucleotides expanded. 
         FIG. 2A  depicts a phosphorothioate antisense oligonucleotide (PS ASO) sequence with end nucleotides expanded. 
         FIG. 2B  depicts a fully expanded phosphorothioate antisense oligonucleotide (PS ASO) sequence. 
         FIG. 3  depicts methods used to quantify skipped DMD mRNA in total RNA using Taqman qPCR. 
         FIG. 4  depicts a chromatogram of anti-CD71 mAb-PMO reaction mixture produced with hydrophobic interaction chromatography (HIC) method 2. 
         FIG. 5A  depicts a chromatogram of anti-CD71 mAb produced using size exclusion chromatography (SEC) method 1. 
         FIG. 5B  depicts a chromatogram of anti-CD71 mAb-PMO DAR 1,2 produced using size exclusion chromatography (SEC) method 1. 
         FIG. 5C  depicts a chromatogram of anti-CD71 mAb-PMO DAR&gt;2 produced using size exclusion chromatography (SEC) method 1. 
         FIG. 6A  depicts a chromatogram of anti-CD71 mAb produced using hydrophobic interaction chromatography (HIC) method 2. 
         FIG. 6B  depicts a chromatogram of purified anti-CD71 mAb-PMO DAR 1,2 conjugate produced using hydrophobic interaction chromatography (HIC) method 2. 
         FIG. 6C  depicts a chromatogram of purified anti-CD71 mAb-PMO DAR&gt;2 conjugate produced using hydrophobic interaction chromatography (HIC) method 2. 
         FIG. 7A  depicts a chromatogram of fast protein liquid chromatography (FPLC) purification of anti-CD71 Fab-PMO using hydrophobic interaction chromatography (HIC) method 3. 
         FIG. 7B  depicts a chromatogram of anti-CD71 Fab produced using SEC method 1. 
         FIG. 7C  depicts a chromatogram of anti-CD71 Fab-PMO DAR 1 conjugate produced using SEC method 1. 
         FIG. 7D  depicts a chromatogram of anti-CD71 Fab-PMO DAR 2 conjugate produced using SEC method 1. 
         FIG. 7E  depicts a chromatogram of anti-CD71 Fab-PMO DAR 3 conjugate produced using SEC method 1. 
         FIG. 7F  depicts a chromatogram of anti-CD71 Fab produced using HIC method 4. 
         FIG. 7G  depicts a chromatogram of anti-CD71 Fab-PMO DAR 1 conjugate produced using HIC method 4. 
         FIG. 7H  depicts a chromatogram of anti-CD71 Fab-PMO DAR 2 conjugate produced using HIC method 4. 
         FIG. 7I  depicts a chromatogram of anti-CD71 Fab-PMO DAR 3 conjugate produced using HIC method 4. 
         FIG. 8A  depicts a chromatogram of anti-CD71 mAb-PS ASO reaction mixture produced with SAX method 2. 
         FIG. 8B  depicts a chromatogram of anti-CD71 mAb produced using SEC method 1. 
         FIG. 8C  depicts a chromatogram of anti-CD71 mAb-PS ASO DAR 1 conjugate produced using SEC method 1. 
         FIG. 8D  depicts a chromatogram of anti-CD71 mAb-PS ASO DAR 2 conjugate produced using SEC method 1. 
         FIG. 8E  depicts a chromatogram of anti-CD71 mAb-PS ASO DAR 3 conjugate produced using SEC method 1. 
         FIG. 8F  depicts a chromatogram of anti-CD71 mAb-PS ASO DAR 1 conjugate produced using SAX method 2. 
         FIG. 8G  depicts a chromatogram of anti-CD71 mAb-PS ASO DAR 2 conjugate produced using SAX method 2. 
         FIG. 8H  depicts a chromatogram of anti-CD71 mAb-PS ASO DAR 3 conjugate produced using SAX method 2. 
         FIG. 9  depicts an agarose gel from nested PCR detecting exon 23 skipping in differentiated C2C12 cells using PMO and anti-CD71 mAb-PMO conjugate. 
         FIG. 10  depicts an agarose gel from nested PCR detecting exon 23 skipping in differentiated C2C12 cells using PMO, anti-CD71 mAb-PMO, and anti-CD71 Fab-PMO conjugates. 
         FIG. 11  depicts an agarose gel from nested PCR detecting exon 23 skipping in differentiated C2C12 cells PMO, ASO, conjugated anti-CD71 mAb-ASO of DAR1 (“ASC-DAR1”), conjugated anti-CD71 mAb-ASO of DAR2 (“ASC-DAR2”), and conjugated anti-CD71 mAb-ASO of DAR3 (“ASC-DAR3”). 
         FIG. 12A  depicts an agarose gel from nested PCR detecting exon 23 skipping in gastrocnemius muscle of wild-type mice administered a single intravenous injection of anti-CD71 mAb-PMO conjugate. 
         FIG. 12B  is a graph of quantification of PCR products from gastrocnemius muscle. 
         FIG. 12C  is a graph of quantification of in vivo exon skipping using Taqman qPCR from gastrocnemius muscle from wild-type mice. 
         FIG. 13A  depicts an agarose gel from nested PCR detecting exon 23 skipping in heart muscle from wild-type mice after a single intravenous injection. 
         FIG. 13B  is a graph of quantification of PCR products from heart muscle. 
         FIG. 14  depicts sequencing data of DNA fragments from skipped and wild-type PCR products. 
         FIG. 15  illustrates exon skipping activity of exon-skipping PMOs at different lengths targeting exon 45 in the human DMD pre-mRNA in transfected primary human skeletal muscle cells. 
         FIG. 16  illustrates binding of hTfR1.mAb-PMO conjugates to human Transferrin Receptor in vitro. 
         FIG. 17  illustrates exon skipping activity of hTfR1.mAb-PMO conjugates in primary human skeletal muscle cells. 
         FIG. 18  illustrates exon skipping activity of hTfR1.mAb-PMO conjugates in myotubes of primary and immortalized human skeletal muscle cells. 
     
    
    
     DETAILED DESCRIPTION OF THE DISCLOSURE 
     Nucleic acid (e.g., RNAi) therapy is a targeted therapy with high selectivity and specificity. However, in some instances, nucleic acid therapy is also hindered by poor intracellular uptake, insufficient intracellular concentrations in target cells, and low efficacy. To address these issues, various modifications of the nucleic acid composition are explored, such as for example, novel linkers for better stabilizing and/or lower toxicity, optimization of binding moiety for increased target specificity and/or target delivery, and nucleic acid polymer modifications for increased stability and/or reduced off-target effect. 
     In some instances, one such area where oligonucleotide is used is for treating muscular dystrophy. Muscular dystrophy encompasses several diseases that affect the muscle. Duchenne muscular dystrophy is a severe form of muscular dystrophy and caused by mutations in the DMD gene. In some instances, mutations in the DMD gene disrupt the translational reading frame and results in non-functional dystrophin protein. 
     Described herein, in certain embodiments, are methods and compositions relating nucleic acid therapy to induce an insertion, deletion, duplication, or alteration in an incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion, which is used to restore the translational reading frame. In some embodiments, also described herein include methods and compositions for treating a disease or disorder characterized by an incorrectly processed mRNA transcript, in which after removal of an exon, the mRNA is capable of encoding a functional protein, thereby treating the disease or disorder. In additional embodiments, described herein include pharmaceutical compositions and kits for treating the same. 
     RNA Processing 
     RNA has a central role in regulation of gene expression and cell physiology. Proper processing of RNA is important for translational of functional protein. Alterations in RNA processing such as a result of incorrect splicing of RNA can result in disease. For example, mutations in a splice site causes exposure of a premature stop codon, a loss of an exon, or inclusion of an intron. In some instances, alterations in RNA processing results in an insertion, deletion, or duplication. In some instances, alterations in RNA processing results in an insertion, deletion, or duplication of an exon. Alterations in RNA processing, in some cases, results in an insertion, deletion, or duplication of an intron. 
     Exon Skipping 
     Exon skipping is a form of RNA splicing. In some cases, exon skipping occurs when an exon is skipped over or is spliced out of the processed mRNA. As a result of exon skipping, the processed mRNA does not contain the skipped exon. In some instances, exon skipping results in expression of an altered product. 
     In some instances, antisense oligonucleotides (AONs) are used to induce exon skipping. In some instances, AONs are short nucleic acid sequences that bind to specific mRNA or pre-mRNA sequences. For example, AONs bind splice sites or exonic enhancers. In some instances, binding of AONs to specific mRNA or pre-mRNA sequences generates double-stranded regions. In some instances, formation of double-stranded regions occurs at sites where the spliceosome or proteins associated with the spliceosome would normally bind and causes exons to be skipped. In some instances, skipping of exons results in restoration of the transcript reading frame and allows for production of a partially functional protein. 
     Exon Inclusion 
     In some instances, a mutation in RNA results in exon skipping. In some cases, a mutation is at least one of at the splice site, near the splice site, and at a distance from the splice site. In some instances, the mutations result in at least one of inactivating or weakening the splice site, disrupting exon splice enhancer or intron splice enhancer, and creating an exon splice silencer or intron splice enhancer. Mutations in some instances alter RNA secondary structure. In some cases, a mutation alters a RNA secondary structure result in disrupting the accessibility of signals important for exon recognition. 
     In some instances, use of AONs results in inclusion of the skipped exon. In some instances, the AONs bind to at least one of a splice site, a site near a splice site, and a site distant to a splice site. In some cases, AONs bind at site in the RNA to prevent disruption of an exon splice enhancer or intron splice enhancer. In some instances, AONs bind at site in the RNA to prevent creation of an exon splice silencer or intron splice silencer. 
     Indications 
     In some embodiments, a polynucleic acid molecule or a pharmaceutical composition described herein is used for the treatment of a disease or disorder characterized with a defective mRNA. In some embodiments, a polynucleic acid molecule or a pharmaceutical composition described herein is used for the treatment of disease or disorder by inducing an insertion, deletion, duplication, or alteration in an incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion. 
     A large percentage of human protein-coding genes are alternatively spliced. In some instances, a mutation results in improperly spliced or partially spliced mRNA. For example, a mutation is in at least one of a splice site in a protein coding gene, a silencer or enhancer sequence, exonic sequences, or intronic sequences. In some instances, a mutation results in gene dysfunction. In some instances, a mutation results in a disease or disorder. 
     In some instances, a disease or disorder resulting from improperly spliced or partially spliced mRNA includes, but not limited to, a neuromuscular disease, a genetic disease, cancer, a hereditary disease, or a cardiovascular disease. 
     In some instances, genetic diseases or disorders include an autosomal dominant disorder, an autosomal recessive disorder, X-linked dominant disorder, X-linked recessive disorder, Y-linked disorder, mitochondrial disease, or multifactorial or polygenic disorder. 
     In some instances, cardiovascular disease such as hypercholesterolemia results from improperly spliced or partially spliced mRNA. In hypercholesterolemia, it has been shown that a single nucleotide polymorphism in exon 12 of the low density lipoprotein receptor (LDLR) promotes exon skipping. 
     In some instances, improperly spliced or partially spliced miRNA results in cancer. For example, improperly spliced or partially spliced EmRNA affects cellular processes involved in cancer including, but not limited to, proliferation, motility, and drug response. In some instances is a solid cancer or a hematologic cancer. In some instances, the cancer is bladder cancer, lung cancer, brain cancer, melanoma, breast cancer, Non-Hodgkin lymphoma, cervical cancer, ovarian cancer, colorectal cancer, pancreatic cancer, esophageal cancer, prostate cancer, kidney cancer, skin cancer, leukemia, thyroid cancer, liver cancer, or uterine cancer. 
     Improperly spliced or partially spliced mRNA in some instances causes a neuromuscular disease or disorder. Exemplary neuromuscular diseases include muscular dystrophy such as Duchenne muscular dystrophy, Becker muscular dystrophy, facioscapulohumeral muscular dystrophy, congenital muscular dystrophy, or myotonic dystrophy. In some instances, muscular dystrophy is genetic. In some instances, muscular dystrophy is caused by a spontaneous mutation. Becker muscular dystrophy and Duchenne muscular dystrophy have been shown to involve mutations in the DMD gene, which encodes the protein dystrophin. Facioscapulohumeral muscular dystrophy has been shown to involve mutations in double homeobox, 4 (DUX4) gene. 
     In some instances, improperly spliced or partially spliced mRNA causes Duchenne muscular dystrophy. Duchenne muscular dystrophy results in severe muscle weakness and is caused by mutations in the DMD gene that abolishes the production of functional dystrophin. In some instances, Duchenne muscular dystrophy is a result of a mutation in an exon in the DMD gene. In some instances, Duchenne muscular dystrophy is a result of a mutation in at least one of exon 1, 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78 and 79 in the DMD gene. In some instances, Duchenne muscular dystrophy is a result of a mutation in at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 in the DMD gene. In some instances, Duchenne muscular dystrophy is a result of a mutation in at least one of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, and 55 in the DMD gene. In some instances, multiple exons are mutated. For example, mutation of exons 48-50 is common in Duchenne muscular dystrophy patients. In some instances, Duchenne muscular dystrophy is a result of mutation of exon 51. In some instances, Duchenne muscular dystrophy is a result of mutation of exon 23. In some instances, a mutation involves a deletion of one or multiple exons. In some instances, a mutation involves a duplication of one or multiple exons. In some instances, a mutation involves a point mutation in an exon. For example, it has been shown that some patients have a nonsense point mutation in exon 51 of the DMD gene. 
     In some instances, a polynucleic acid molecule or a pharmaceutical composition described herein is used for the treatment of muscular dystrophy. In some instances, a polynucleic acid molecule or a pharmaceutical composition described herein is used for the treatment of Duchenne muscular dystrophy, Becker muscular dystrophy, facioscapulohumeral muscular dystrophy, congenital muscular dystrophy, or myotonic dystrophy. In some instances, a polynucleic acid molecule or a pharmaceutical composition described herein is used for the treatment of Duchenne muscular dystrophy. 
     Polynucleic Acid Molecule 
     In some embodiments, a polynucleic acid molecule described herein that induces an insertion, deletion, duplication, or alteration in an incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion. In some instances, the polynucleic acid molecule restores the translational reading frame. In some instances, the polynucleic acid molecule results in a functional and truncated protein. 
     In some instances, a polynucleic acid molecule targets an mRNA sequence. In some instances, the polynucleic acid molecule targets a splice site. In some instances, the polynucleic acid molecule targets a cis-regulatory element. In some instances, the polynucleic molecule targets a trans-regulatory element. In some instances, the polynucleic acid molecule targets exonic splice enhancers or intronic splice enhancers. In some instances, the polynucleic acid molecule targets exonic splice silencers or intronic splice silencers. 
     In some instances, a polynucleic acid molecule targets a sequence found in introns or exons. For example, the polynucleic acid molecule targets a sequence found in an exon that mediates splicing of said exon. In some instances, the polynucleic acid molecule targets an exon recognition sequence. In some instances, the polynucleic acid molecule targets a sequence upstream of an exon. In some instances, the polynucleic acid molecule targets a sequence downstream of an exon. 
     As described above, a polynucleic acid molecule targets an incorrectly processed mRNA transcript which results in a disease or disorder not limited to a neuromuscular disease, a genetic disease, cancer, a hereditary disease, or a cardiovascular disease. In some cases, a polynucleic acid molecule targets an incorrectly processed mRNA transcript which results in a neuromuscular disease or disorder. In some cases, a neuromuscular disease or disorder is Duchenne muscular dystrophy, Becker muscular dystrophy, facioscapulohumeral muscular dystrophy, congenital muscular dystrophy, or myotonic dystrophy. In some cases, a polynucleic acid molecule targets an incorrectly processed mRNA transcript which results in Duchenne muscular dystrophy, Becker muscular dystrophy, facioscapulohumeral muscular dystrophy, congenital muscular dystrophy, or myotonic dystrophy. In some cases, a polynucleic acid molecule targets an incorrectly processed mRNA transcript which results in Duchenne muscular dystrophy. 
     In some instances, a polynucleic acid molecule targets an exon that is mutated in the DMD gene that causes Duchenne muscular dystrophy. Exemplary exons that are mutated in the DMD gene that causes Duchenne muscular dystrophy include, but not limited to, exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, and 78. In some instances, the polynucleic acid molecule targets a sequence adjacent to a mutated exon. For example, if there is a deletion of exon 50, the polynucleic acid molecule targets a sequence in exon 51 so that exon 51 is skipped. In another instance, if there is a mutation in exon 23, the polynucleic acid molecule targets a sequence in exon 22 so that exon 23 is skipped. 
     In some instances, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, or 78 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, or 63 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 8 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 23 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 35 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 43 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 44 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 45 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 48 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 49 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 50 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 51 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 52 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 53 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a region that is at the exon-intron junction of exon 55 of the DMD gene. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is at either the 5′ intron-exon junction or the 3′ exon-intron junction of at least one of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, and 78 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is at either the 5′ intron-exon junction or the 3′ exon-intron junction of at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is at either the 5′ intron-exon junction or the 3′ exon-intron junction of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. 
     In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of at least one of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, and 78 of the DMD gene (e.g., the 5′ intron-exon junction of exon 3 is the junction intron 2-exon 3). In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 of the DMD gene (e.g., the 5′ intron-exon junction of exon 3 is the junction intron 2-exon 3). In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 8 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 23 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 35 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 43 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 44 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 45 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 50 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 51 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 52 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 53 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 5′ intron-exon junction of exon 55 of the DMD gene. 
     In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of at least one of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, and 78 of the DMD gene (e.g., the 3′ exon-intron junction of exon 3 is the junction exon 3-intron 3). In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 of the DMD gene (e.g., the 3′ exon-intron junction of exon 3 is the junction exon 3-intron 3). In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 8 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 23 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 35 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 43 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 44 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 45 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 50 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 51 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 52 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 53 of the DMD gene. In some cases, the polynucleic acid molecule hybridizes to a target region that is at the 3′ exon-intron junction of exon 55 of the DMD gene. 
     In some instances, a polynucleic acid molecule described herein targets a splice site of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, and 78 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a splice site of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, or 63 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a splice site of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a splice site of exon 8 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a splice site of exon 23 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a splice site of exon 35 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a splice site of exon 43 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a splice site of exon 44 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a splice site of exon 45 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a splice site of exon 48 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a splice site of exon 49 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a splice site of exon 50 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a splice site of exon 51 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a splice site of exon 52 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a splice site of exon 53 of the DMD gene. In some cases, a polynucleic acid molecule described herein targets a splice site of exon 55 of the DMD gene. As used herein, a splice site includes a canonical splice site, a cryptic splice site or an alternative splice site that is capable of inducing an insertion, deletion, duplication, or alteration in an incorrectly spliced mRNA transcript to induce exon skipping or exon inclusion. 
     In some embodiments, a polynucleic acid molecule described herein target a partially spliced mRNA sequence comprising additional exons involved in Duchenne muscular dystrophy such as exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, or 63. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is proximal to the exon-intron junction. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, or 78 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, or 63 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 8 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 23 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 35 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 43 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 44 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 45 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 48 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 49 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 50 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 51 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 52 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 53 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nt, 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt upstream (or from the 5′) of exon 55 of the DMD gene. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is upstream (or 5′) to at least one of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, and 78 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is upstream (or 5′) to at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is upstream (or 5′) to at least one of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp upstream (or 5′) to at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 of the DMD gene. 
     In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, or 78 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, or 63 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 8 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 23 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 35 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 43 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 44 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 45 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 48 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 49 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 50 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 51 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 52 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 53 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets a region at least 1000 nucleotides (nt), 500 nt, 400 nt, 300 nt, 200 nt, 100 nt, 80 nt, 60 nt, 50 nt, 40 nt, 30 nt, 20 nt, 10 nt, or 5 nt downstream (or from the 3′) of exon 55 of the DMD gene. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is downstream (or 3′) to at least one of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, and 78 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is downstream (or 3′) to at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp downstream (or 3′) to at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp downstream (or 3′) to at least one of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. 
     In some instances, a polynucleic acid molecule described herein targets an internal region within exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, or 78 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, or 63 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 8 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 23 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 35 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 43 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 44 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 45 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 48 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 49 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 50 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 51 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 52 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 53 of the DMD gene. In some instances, a polynucleic acid molecule described herein targets an internal region within exon 55 of the DMD gene. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is within at least one of exon 2, 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, and 78 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is within at least one of exon 3, 4, 5, 6, 7, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, and 63 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is within at least one of exon 8, 23, 35, 43, 44, 45, 50, 51, 52, 53, or 55 of the DMD gene. 
     In some embodiments, a polynucleic acid molecule described herein targets a partially spliced mRNA sequence comprising exon 44 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is upstream (or 5′) to exon 44. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp upstream (or 5′) to exon 44. In some instances, the polynucleic acid molecule hybridizes to a target region that is downstream (or 3′) to exon 44. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp downstream (or 3′) to exon 44. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is within exon 44 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is at either the 5′ intron-exon 44 junction or the 3′ exon 44-intron junction. 
     In some embodiments, a polynucleic acid molecule described herein targets a partially spliced mRNA sequence comprising exon 45 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is upstream (or 5′) to exon 45. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp upstream (or 5′) to exon 45. In some instances, the polynucleic acid molecule hybridizes to a target region that is downstream (or 3′) to exon 45. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp downstream (or 3′) to exon 45. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is within exon 45 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is at either the 5′ intron-exon 45 junction or the 3′ exon 45-intron junction. 
     In some embodiments, a polynucleic acid molecule described herein targets a partially spliced mRNA sequence comprising exon 51 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is upstream (or 5′) to exon 51. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp upstream (or 5′) to exon 51. In some instances, the polynucleic acid molecule hybridizes to a target region that is downstream (or 3′) to exon 51. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp downstream (or 3′) to exon 51. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is within exon 51 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is at either the 5′ intron-exon 51 junction or the 3′ exon 51-intron junction. 
     In some embodiments, a polynucleic acid molecule described herein targets a partially spliced mRNA sequence comprising exon 53 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is upstream (or 5′) to exon 53. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp upstream (or 5′) to exon 53. In some instances, the polynucleic acid molecule hybridizes to a target region that is downstream (or 3′) to exon 53. In some instances, the polynucleic acid molecule hybridizes to a target region that is about 5, 10, 15, 20, 50, 100, 200, 300, 400 or 500 bp downstream (or 3′) to exon 53. 
     In some instances, the polynucleic acid molecule hybridizes to a target region that is within exon 53 of the DMD gene. In some instances, the polynucleic acid molecule hybridizes to a target region that is at either the 5′ intron-exon 53 junction or the 3′ exon 53-intron junction. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to a target sequence of interest. In some embodiments, the polynucleic acid molecule consists of a target sequence of interest. 
     In some embodiments, the polynucleic acid molecule comprises a first polynucleotide and a second polynucleotide. In some instances, the first polynucleotide comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a target sequence of interest. In some cases, the second polynucleotide comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a target sequence of interest. In some cases, the polynucleic acid molecule comprises a first polynucleotide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a target sequence of interest and a second polynucleotide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to a target sequence of interest. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to SEQ ID NOs: 964-1285. In some embodiments, the polynucleic acid molecule consists of SEQ ID NOs: 964-1285. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to SEQ ID NOs: 1056-1094. In some embodiments, the polynucleic acid molecule consists of SEQ ID NOs: 1056-1094. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to SEQ ID NOs: 1147-1162. In some embodiments, the polynucleic acid molecule consists of SEQ ID NOs: 1147-1162. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to SEQ ID NOs: 1173-1211. In some embodiments, the polynucleic acid molecule consists of SEQ ID NOs: 1173-1211. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to SEQ ID NOs: 1056-1076. In some embodiments, the polynucleic acid molecule consists of SEQ ID NOs: 1056-1076. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to SEQ ID NOs: 1077-1094. In some embodiments, the polynucleic acid molecule consists of SEQ ID NOs: 1077-1094. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to SEQ ID NOs: 1056-1058. In some embodiments, the polynucleic acid molecule consists of SEQ ID NOs: 1056-1058. 
     In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 50% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 60% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 70% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 75% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 80% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 85% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 90% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 95% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 96% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 97% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 98% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule comprises a sequence having at least 99% sequence identity to SEQ ID NOs: 1087-1089. In some embodiments, the polynucleic acid molecule consists of SEQ ID NOs: 1087-1089. 
     In some embodiments, the polynucleic acid molecule at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 964-1285. In some instances, the polynucleic acid molecule at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1056-1094, 1147-1162, or 1173-1211. In some instances, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1056-1076. In some instances, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1077-1094. In some instances, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1147-1162. In some instances, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1173-1211. In some instances, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1056-1058. In some instances, the polynucleic acid molecule comprises at least 10, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or more contiguous bases of a base sequence selected from SEQ ID NOs: 1087-1089. In some cases, the polynucleic acid molecule further comprises 1, 2, 3, or 4 mismatches. 
     In some embodiments, the polynucleic acid molecule comprises a guide strand and a passenger strand. In some instances, the guide strand comprises a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 964-1285. In some cases, the guide strand comprises a sequence selected from SEQ ID NOs: 964-1285. 
     In some embodiments, the polynucleic acid molecule described herein comprises RNA or DNA. In some cases, the polynucleic acid molecule comprises RNA. In some instances, RNA comprises short interfering RNA (siRNA), short hairpin RNA (shRNA), microRNA (miRNA), double-stranded RNA (dsRNA), transfer RNA (tRNA), ribosomal RNA (rRNA), or heterogeneous nuclear RNA (hnRNA). In some instances, RNA comprises shRNA. In some instances, RNA comprises miRNA. In some instances, RNA comprises dsRNA. In some instances, RNA comprises tRNA. In some instances, RNA comprises rRNA. In some instances, RNA comprises hnRNA. In some instances, the RNA comprises siRNA. In some instances, the polynucleic acid molecule comprises siRNA. In some instances, the polynucleic acid molecule is an antisense oligonucleotide (ASO). 
     In some embodiments, the polynucleic acid molecule is from about 10 to about 50 nucleotides in length. In some instances, the polynucleic acid molecule is from about 10 to about 30, from about 15 to about 30, from about 18 to about 30, from about 18 to about 25, form about 18 to about 24, from about 19 to about 23, from about 19 to about 30, from about 19 to about 25, form about 19 to about 24, from about 19 to about 23, from about 20 to about 30, from about 20 to about 25, from about 20 to about 24, from about 20 to about 23, or from about 20 to about 22 nucleotides in length. 
     In some embodiments, the polynucleic acid molecule is about 50 nucleotides in length. In some instances, the polynucleic acid molecule is about 45 nucleotides in length. In some instances, the polynucleic acid molecule is about 40 nucleotides in length. In some instances, the polynucleic acid molecule is about 35 nucleotides in length. In some instances, the polynucleic acid molecule is about 30 nucleotides in length. In some instances, the polynucleic acid molecule is about 25 nucleotides in length. In some instances, the polynucleic acid molecule is about 20 nucleotides in length. In some instances, the polynucleic acid molecule is about 19 nucleotides in length. In some instances, the polynucleic acid molecule is about 18 nucleotides in length. In some instances, the polynucleic acid molecule is about 17 nucleotides in length. In some instances, the polynucleic acid molecule is about 16 nucleotides in length. In some instances, the polynucleic acid molecule is about 15 nucleotides in length. In some instances, the polynucleic acid molecule is about 14 nucleotides in length. In some instances, the polynucleic acid molecule is about 13 nucleotides in length. In some instances, the polynucleic acid molecule is about 12 nucleotides in length. In some instances, the polynucleic acid molecule is about 11 nucleotides in length. In some instances, the polynucleic acid molecule is about 10 nucleotides in length. In some instances, the polynucleic acid molecule is between about 10 and about 50 nucleotides in length. In some instances, the polynucleic acid molecule is between about 10 and about 45 nucleotides in length. In some instances, the polynucleic acid molecule is between about 10 and about 40 nucleotides in length. In some instances, the polynucleic acid molecule is between about 10 and about 35 nucleotides in length. In some instances, the polynucleic acid molecule is between about 10 and about 30 nucleotides in length. In some instances, the polynucleic acid molecule is between about 10 and about 25 nucleotides in length. In some instances, the polynucleic acid molecule is between about 10 and about 20 nucleotides in length. In some instances, the polynucleic acid molecule is between about 15 and about 25 nucleotides in length. In some instances, the polynucleic acid molecule is between about 15 and about 30 nucleotides in length. In some instances, the polynucleic acid molecule is between about 12 and about 30 nucleotides in length. In some instances, the polynucleic acid molecule is between about 19 and about 30 nucleotides in length. In some instances, the polynucleic acid molecule is between about 20 and about 30 nucleotides in length. In some instances, the polynucleic acid molecule is between about 19 and about 25 nucleotides in length. In some instances, the polynucleic acid molecule is between about 20 and about 25 nucleotides in length. 
     In some embodiments, the polynucleic acid molecule is at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, or 50 nucleotides in length. In some instances, the polynucleic acid molecule is at least 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length. In some instances, the polynucleic acid molecule is at least 15 nucleotides in length. In some instances, the polynucleic acid molecule is at least 18 nucleotides in length. In some instances, the polynucleic acid molecule is at least 19 nucleotides in length. In some instances, the polynucleic acid molecule is at least 20 nucleotides in length. In some instances, the polynucleic acid molecule is at least 21 nucleotides in length. In some instances, the polynucleic acid molecule is at least 22 nucleotides in length. In some instances, the polynucleic acid molecule is at least 23 nucleotides in length. In some instances, the polynucleic acid molecule is at least 24 nucleotides in length. In some instances, the polynucleic acid molecule is at least 25 nucleotides in length. In some instances, the polynucleic acid molecule is at least 30 nucleotides in length. 
     In some embodiments, the polynucleic acid molecule is about 50 nucleotides in length. In some instances, the polynucleic acid molecule is about 45 nucleotides in length. In some instances, the polynucleic acid molecule is about 40 nucleotides in length. In some instances, the polynucleic acid molecule is about 35 nucleotides in length. In some instances, the polynucleic acid molecule is about 30 nucleotides in length. In some instances, the polynucleic acid molecule is about 29 nucleotides in length. In some instances, the polynucleic acid molecule is about 28 nucleotides in length. In some instances, the polynucleic acid molecule is about 27 nucleotides in length. In some instances, the polynucleic acid molecule is about 26 nucleotides in length. In some instances, the polynucleic acid molecule is about 25 nucleotides in length. In some instances, the polynucleic acid molecule is about 24 nucleotides in length. In some instances, the polynucleic acid molecule is about 23 nucleotides in length. In some instances, the polynucleic acid molecule is about 22 nucleotides in length. In some instances, the polynucleic acid molecule is about 21 nucleotides in length. In some instances, the polynucleic acid molecule is about 20 nucleotides in length. In some instances, the polynucleic acid molecule is about 19 nucleotides in length. In some instances, the polynucleic acid molecule is about 18 nucleotides in length. In some instances, the polynucleic acid molecule is about 17 nucleotides in length. In some instances, the polynucleic acid molecule is about 16 nucleotides in length. In some instances, the polynucleic acid molecule is about 15 nucleotides in length. In some instances, the polynucleic acid molecule is about 14 nucleotides in length. In some instances, the polynucleic acid molecule is about 13 nucleotides in length. In some instances, the polynucleic acid molecule is about 12 nucleotides in length. In some instances, the polynucleic acid molecule is about 11 nucleotides in length. In some instances, the polynucleic acid molecule is about 10 nucleotides in length. 
     In some embodiments, the polynucleic acid molecule comprises a first polynucleotide. In some instances, the polynucleic acid molecule comprises a second polynucleotide. In some instances, the polynucleic acid molecule comprises a first polynucleotide and a second polynucleotide. In some instances, the first polynucleotide is a sense strand or passenger strand. In some instances, the second polynucleotide is an antisense strand or guide strand. 
     In some embodiments, the polynucleic acid molecule is a first polynucleotide. In some embodiments, the first polynucleotide is from about 10 to about 50 nucleotides in length. In some instances, the first polynucleotide is from about 10 to about 30, from about 15 to about 30, from about 18 to about 30, from about 18 to about 25, form about 18 to about 24, from about 19 to about 23, from about 19 to about 30, from about 19 to about 25, form about 19 to about 24, from about 19 to about 23, from about 20 to about 30, from about 20 to about 25, from about 20 to about 24, from about 20 to about 23, or from about 20 to about 22 nucleotides in length. 
     In some instances, the first polynucleotide is about 50 nucleotides in length. In some instances, the first polynucleotide is about 45 nucleotides in length. In some instances, the first polynucleotide is about 40 nucleotides in length. In some instances, the first polynucleotide is about 35 nucleotides in length. In some instances, the first polynucleotide is about 30 nucleotides in length. In some instances, the first polynucleotide is about 25 nucleotides in length. In some instances, the first polynucleotide is about 20 nucleotides in length. In some instances, the first polynucleotide is about 19 nucleotides in length. In some instances, the first polynucleotide is about 18 nucleotides in length. In some instances, the first polynucleotide is about 17 nucleotides in length. In some instances, the first polynucleotide is about 16 nucleotides in length. In some instances, the first polynucleotide is about 15 nucleotides in length. In some instances, the first polynucleotide is about 14 nucleotides in length. In some instances, the first polynucleotide is about 13 nucleotides in length. In some instances, the first polynucleotide is about 12 nucleotides in length. In some instances, the first polynucleotide is about 11 nucleotides in length. In some instances, the first polynucleotide is about 10 nucleotides in length. In some instances, the first polynucleotide is between about 10 and about 50 nucleotides in length. In some instances, the first polynucleotide is between about 10 and about 45 nucleotides in length. In some instances, the first polynucleotide is between about 10 and about 40 nucleotides in length. In some instances, the first polynucleotide is between about 10 and about 35 nucleotides in length. In some instances, the first polynucleotide is between about 10 and about 30 nucleotides in length. In some instances, the first polynucleotide is between about 10 and about 25 nucleotides in length. In some instances, the first polynucleotide is between about 10 and about 20 nucleotides in length. In some instances, the first polynucleotide is between about 15 and about 25 nucleotides in length. In some instances, the first polynucleotide is between about 15 and about 30 nucleotides in length. In some instances, the first polynucleotide is between about 12 and about 30 nucleotides in length. 
     In some embodiments, the polynucleic acid molecule is a second polynucleotide. In some embodiments, the second polynucleotide is from about 10 to about 50 nucleotides in length. In some instances, the second polynucleotide is from about 10 to about 30, from about 15 to about 30, from about 18 to about 30, from about 18 to about 25, form about 18 to about 24, from about 19 to about 23, from about 19 to about 30, from about 19 to about 25, form about 19 to about 24, from about 19 to about 23, from about 20 to about 30, from about 20 to about 25, from about 20 to about 24, from about 20 to about 23, or from about 20 to about 22 nucleotides in length. 
     In some instances, the second polynucleotide is about 50 nucleotides in length. In some instances, the second polynucleotide is about 45 nucleotides in length. In some instances, the second polynucleotide is about 40 nucleotides in length. In some instances, the second polynucleotide is about 35 nucleotides in length. In some instances, the second polynucleotide is about 30 nucleotides in length. In some instances, the second polynucleotide is about 25 nucleotides in length. In some instances, the second polynucleotide is about 20 nucleotides in length. In some instances, the second polynucleotide is about 19 nucleotides in length. In some instances, the second polynucleotide is about 18 nucleotides in length. In some instances, the second polynucleotide is about 17 nucleotides in length. In some instances, the second polynucleotide is about 16 nucleotides in length. In some instances, the second polynucleotide is about 15 nucleotides in length. In some instances, the second polynucleotide is about 14 nucleotides in length. In some instances, the second polynucleotide is about 13 nucleotides in length. In some instances, the second polynucleotide is about 12 nucleotides in length. In some instances, the second polynucleotide is about 11 nucleotides in length. In some instances, the second polynucleotide is about 10 nucleotides in length. In some instances, the second polynucleotide is between about 10 and about 50 nucleotides in length. In some instances, the second polynucleotide is between about 10 and about 45 nucleotides in length. In some instances, the second polynucleotide is between about 10 and about 40 nucleotides in length. In some instances, the second polynucleotide is between about 10 and about 35 nucleotides in length. In some instances, the second polynucleotide is between about 10 and about 30 nucleotides in length. In some instances, the second polynucleotide is between about 10 and about 25 nucleotides in length. In some instances, the second polynucleotide is between about 10 and about 20 nucleotides in length. In some instances, the second polynucleotide is between about 15 and about 25 nucleotides in length. In some instances, the second polynucleotide is between about 15 and about 30 nucleotides in length. In some instances, the second polynucleotide is between about 12 and about 30 nucleotides in length. 
     In some embodiments, the polynucleic acid molecule comprises a first polynucleotide and a second polynucleotide. In some instances, the polynucleic acid molecule further comprises a blunt terminus, an overhang, or a combination thereof. In some instances, the blunt terminus is a 5′ blunt terminus, a 3′ blunt terminus, or both. In some cases, the overhang is a 5′ overhang, 3′ overhang, or both. In some cases, the overhang comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 non-base pairing nucleotides. In some cases, the overhang comprises 1, 2, 3, 4, 5, or 6 non-base pairing nucleotides. In some cases, the overhang comprises 1, 2, 3, or 4 non-base pairing nucleotides. In some cases, the overhang comprises 1 non-base pairing nucleotide. In some cases, the overhang comprises 2 non-base pairing nucleotides. In some cases, the overhang comprises 3 non-base pairing nucleotides. In some cases, the overhang comprises 4 non-base pairing nucleotides. 
     In some embodiments, the sequence of the polynucleic acid molecule is at least 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.5% complementary to a target sequence described herein. In some embodiments, the sequence of the polynucleic acid molecule is at least 50% complementary to a target sequence described herein. In some embodiments, the sequence of the polynucleic acid molecule is at least 60% complementary to a target sequence described herein. In some embodiments, the sequence of the polynucleic acid molecule is at least 70% complementary to a target sequence described herein. In some embodiments, the sequence of the polynucleic acid molecule is at least 80% complementary to a target sequence described herein. In some embodiments, the sequence of the polynucleic acid molecule is at least 90% complementary to a target sequence described herein. In some embodiments, the sequence of the polynucleic acid molecule is at least 95% complementary to a target sequence described herein. In some embodiments, the sequence of the polynucleic acid molecule is at least 99% complementary to a target sequence described herein. In some instances, the sequence of the polynucleic acid molecule is 100% complementary to a target sequence described herein. 
     In some embodiments, the sequence of the polynucleic acid molecule has 5 or less mismatches to a target sequence described herein. In some embodiments, the sequence of the polynucleic acid molecule has 4 or less mismatches to a target sequence described herein. In some instances, the sequence of the polynucleic acid molecule has 3 or less mismatches to a target sequence described herein. In some cases, the sequence of the polynucleic acid molecule has 2 or less mismatches to a target sequence described herein. In some cases, the sequence of the polynucleic acid molecule has 1 or less mismatches to a target sequence described herein. 
     In some embodiments, the specificity of the polynucleic acid molecule that hybridizes to a target sequence described herein is a 95%, 98%, 99%, 99.5% or 100% sequence complementarity of the polynucleic acid molecule to a target sequence. In some instances, the hybridization is a high stringent hybridization condition. 
     In some embodiments, the polynucleic acid molecule has reduced off-target effect. In some instances, “off-target” or “off-target effects” refer to any instance in which a polynucleic acid polymer directed against a given target causes an unintended effect by interacting either directly or indirectly with another mRNA sequence, a DNA sequence or a cellular protein or other moiety. In some instances, an “off-target effect” occurs when there is a simultaneous degradation of other transcripts due to partial homology or complementarity between that other transcript and the sense and/or antisense strand of the polynucleic acid molecule. 
     In some embodiments, the polynucleic acid molecule comprises natural or synthetic or artificial nucleotide analogues or bases. In some cases, the polynucleic acid molecule comprises combinations of DNA, RNA and/or nucleotide analogues. In some instances, the synthetic or artificial nucleotide analogues or bases comprise modifications at one or more of ribose moiety, phosphate moiety, nucleoside moiety, or a combination thereof. 
     In some embodiments, nucleotide analogues or artificial nucleotide base comprise a nucleic acid with a modification at a 2′ hydroxyl group of the ribose moiety. In some instances, the modification includes an H, OR, R, halo, SH, SR, NH2, NHR, NR2, or CN, wherein R is an alkyl moiety. Exemplary alkyl moiety includes, but is not limited to, halogens, sulfurs, thiols, thioethers, thioesters, amines (primary, secondary, or tertiary), amides, ethers, esters, alcohols and oxygen. In some instances, the alkyl moiety further comprises a modification. In some instances, the modification comprises an azo group, a keto group, an aldehyde group, a carboxyl group, a nitro group, a nitroso, group, a nitrile group, a heterocycle (e.g., imidazole, hydrazino or hydroxylamino) group, an isocyanate or cyanate group, or a sulfur containing group (e.g., sulfoxide, sulfone, sulfide, or disulfide). In some instances, the alkyl moiety further comprises a hetero substitution. In some instances, the carbon of the heterocyclic group is substituted by a nitrogen, oxygen or sulfur. In some instances, the heterocyclic substitution includes but is not limited to, morpholino, imidazole, and pyrrolidino. 
     In some instances, the modification at the 2′ hydroxyl group is a 2′-O-methyl modification or a 2′-O-methoxyethyl (2′-O-MOE) modification. In some cases, the 2′-O-methyl modification adds a methyl group to the 2′ hydroxyl group of the ribose moiety whereas the 2′O-methoxyethyl modification adds a methoxyethyl group to the 2′ hydroxyl group of the ribose moiety. Exemplary chemical structures of a 2′-O-methyl modification of an adenosine molecule and 2′O-methoxyethyl modification of an uridine are illustrated below. 
     
       
         
         
             
             
         
       
     
     In some instances, the modification at the 2′ hydroxyl group is a 2′-O-aminopropyl modification in which an extended amine group comprising a propyl linker binds the amine group to the 2′ oxygen. In some instances, this modification neutralizes the phosphate derived overall negative charge of the oligonucleotide molecule by introducing one positive charge from the amine group per sugar and thereby improves cellular uptake properties due to its zwitterionic properties. An exemplary chemical structure of a 2′-O-aminopropyl nucleoside phosphoramidite is illustrated below. 
     
       
         
         
             
             
         
       
     
     In some instances, the modification at the 2′ hydroxyl group is a locked or bridged ribose modification (e.g., locked nucleic acid or LNA) in which the oxygen molecule bound at the 2′ carbon is linked to the 4′ carbon by a methylene group, thus forming a 2′-C,4′-C-oxy-methylene-linked bicyclic ribonucleotide monomer. Exemplary representations of the chemical structure of LNA are illustrated below. The representation shown to the left highlights the chemical connectivities of an LNA monomer. The representation shown to the right highlights the locked 3′-endo ( 3 E) conformation of the furanose ring of an LNA monomer. 
     
       
         
         
             
             
         
       
     
     In some instances, the modification at the 2′ hydroxyl group comprises ethylene nucleic acids (ENA) such as for example 2′-4′-ethylene-bridged nucleic acid, which locks the sugar conformation into a C 3 ′-endo sugar puckering conformation. ENA are part of the bridged nucleic acids class of modified nucleic acids that also comprises LNA. Exemplary chemical structures of the ENA and bridged nucleic acids are illustrated below. 
     
       
         
         
             
             
         
       
     
     In some embodiments, additional modifications at the 2′ hydroxyl group include 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA). 
     In some embodiments, nucleotide analogues comprise modified bases such as, but not limited to, 5-propynyluridine, 5-propynylcytidine, 6-methyladenine, 6-methylguanine, N, N,-dimethyladenine, 2-propyladenine, 2propylguanine, 2-aminoadenine, 1-methylinosine, 3-methyluridine, 5-methylcytidine, 5-methyluridine and other nucleotides having a modification at the 5 position, 5-(2-amino) propyl uridine, 5-halocytidine, 5-halouridine, 4-acetylcytidine, 1-methyladenosine, 2-methyladenosine, 3-methylcytidine, 6-methyluridine, 2-methylguanosine, 7-methylguanosine, 2, 2-dimethylguanosine, 5-methylaminoethyluridine, 5-methyloxyuridine, deazanucleotides such as 7-deaza-adenosine, 6-azouridine, 6-azocytidine, 6-azothymidine, 5-methyl-2-thiouridine, other thio bases such as 2-thiouridine and 4-thiouridine and 2-thiocytidine, dihydrouridine, pseudouridine, queuosine, archaeosine, naphthyl and substituted naphthyl groups, any O- and N-alkylated purines and pyrimidines such as N6-methyladenosine, 5-methylcarbonylmethyluridine, uridine 5-oxyacetic acid, pyridine-4-one, pyridine-2-one, phenyl and modified phenyl groups such as aminophenol or 2,4, 6-trimethoxy benzene, modified cytosines that act as G-clamp nucleotides, 8-substituted adenines and guanines, 5-substituted uracils and thymines, azapyrimidines, carboxyhydroxyalkyl nucleotides, carboxyalkylaminoalkyi nucleotides, and alkylcarbonylalkylated nucleotides. Modified nucleotides also include those nucleotides that are modified with respect to the sugar moiety, as well as nucleotides having sugars or analogs thereof that are not ribosyl. For example, the sugar moieties, in some cases are or be based on, mannoses, arabinoses, glucopyranoses, galactopyranoses, 4′-thioribose, and other sugars, heterocycles, or carbocycles. The term nucleotide also includes what are known in the art as universal bases. By way of example, universal bases include but are not limited to 3-nitropyrrole, 5-nitroindole, or nebularine. 
     In some embodiments, nucleotide analogues further comprise morpholinos, peptide nucleic acids (PNAs), methylphosphonate nucleotides, thiolphosphonate nucleotides, 2′-fluoro N3-P5′-phosphoramidites, 1′,5′-anhydrohexitol nucleic acids (HNAs), or a combination thereof. Morpholino or phosphorodiamidate morpholino oligo (PMO) comprises synthetic molecules whose structure mimics natural nucleic acid structure by deviates from the normal sugar and phosphate structures. In some instances, the five member ribose ring is substituted with a six member morpholino ring containing four carbons, one nitrogen and one oxygen. In some cases, the ribose monomers are linked by a phosphordiamidate group instead of a phosphate group. In such cases, the backbone alterations remove all positive and negative charges making morpholinos neutral molecules capable of crossing cellular membranes without the aid of cellular delivery agents such as those used by charged oligonucleotides. 
     
       
         
         
             
             
         
       
     
     In some embodiments, peptide nucleic acid (PNA) does not contain sugar ring or phosphate linkage and the bases are attached and appropriately spaced by oligoglycine-like molecules, therefore, eliminating a backbone charge. 
     
       
         
         
             
             
         
       
     
     In some embodiments, one or more modifications optionally occur at the internucleotide linkage. In some instances, modified internucleotide linkage include, but is not limited to, phosphorothioates, phosphorodithioates, methylphosphonates, 5′-alkylenephosphonates, 5′-methylphosphonate, 3′-alkylene phosphonates, borontrifluoridates, borano phosphate esters and selenophosphates of 3′-5′linkage or 2′-5′linkage, phosphotriesters, thionoalkylphosphotriesters, hydrogen phosphonate linkages, alkyl phosphonates, alkylphosphonothioates, arylphosphonothioates, phosphoroselenoates, phosphorodiselenoates, phosphinates, phosphoramidates, 3′-alkylphosphoramidates, aminoalkylphosphoramidates, thionophosphoramidates, phosphoropiperazidates, phosphoroanilothioates, phosphoroanilidates, ketones, sulfones, sulfonamides, carbonates, carbamates, methylenehydrazos, methylenedimethylhydrazos, formacetals, thioformacetals, oximes, methyleneiminos, methylenemethyliminos, thioamidates, linkages with riboacetyl groups, aminoethyl glycine, silyl or siloxane linkages, alkyl or cycloalkyl linkages with or without heteroatoms of, for example, 1 to 10 carbons that are saturated or unsaturated and/or substituted and/or contain heteroatoms, linkages with morpholino structures, amides, polyamides wherein the bases are attached to the aza nitrogens of the backbone directly or indirectly, and combinations thereof. Phosphorothioate antisene oligonucleotides (PS ASO) are antisense oligonucleotides comprising a phosphorothioate linkage. An exemplary PS ASO is illustrated below. 
     
       
         
         
             
             
         
       
     
     In some instances, the modification is a methyl or thiol modification such as methylphosphonate or thiolphosphonate modification. Exemplary thiolphosphonate nucleotide (left) and methylphosphonate nucleotide (right) are illustrated below. 
     
       
         
         
             
             
         
       
     
     In some instances, a modified nucleotide includes, but is not limited to, 2′-fluoro N3-P5′-phosphoramidites illustrated as: 
     
       
         
         
             
             
         
       
     
     In some instances, a modified nucleotide includes, but is not limited to, hexitol nucleic acid (or 1′,5′-anhydrohexitol nucleic acids (HNA)) illustrated as: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a nucleotide analogue or artificial nucleotide base described above comprises a 5′-vinylphosphonate modified nucleotide nucleic acid with a modification at a 5′ hydroxyl group of the ribose moiety. In some embodiments, the 5′-vinylphosphonate modified nucleotide is selected from the nucleotide provided below, wherein X is O or S; and B is a heterocyclic base moiety. 
     
       
         
         
             
             
         
       
       
         
         
             
             
         
       
     
     In some instances, the modification at the 2′ hydroxyl group is a 2′-O-aminopropyl modification in which an extended amine group comprising a propyl linker binds the amine group to the 2′ oxygen. In some instances, this modification neutralizes the phosphate-derived overall negative charge of the oligonucleotide molecule by introducing one positive charge from the amine group per sugar and thereby improves cellular uptake properties due to its zwitterionic properties. 
     In some instances, the 5′-vinylphosphonate modified nucleotide is further modified at the 2′ hydroxyl group in a locked or bridged ribose modification (e.g., locked nucleic acid or LNA) in which the oxygen molecule bound at the 2′ carbon is linked to the 4′ carbon by a methylene group, thus forming a 2′-C,4′-C-oxy-methylene-linked bicyclic ribonucleotide monomer. Exemplary representations of the chemical structure of 5′-vinylphosphonate modified LNA are illustrated below, wherein X is O or S; B is a heterocyclic base moiety; and J is an internucleotide linking group linking to the adjacent nucleotide of the polynucleotide. 
     
       
         
         
             
             
         
       
     
     In some embodiments, additional modifications at the 2′ hydroxyl group include 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA). 
     In some embodiments, a nucleotide analogue comprises a modified base such as, but not limited to, 5-propynyluridine, 5-propynylcytidine, 6-methyladenine, 6-methylguanine, N, N,-dimethyladenine, 2-propyladenine, 2propylguanine, 2-aminoadenine, 1-methylinosine, 3-methyluridine, 5-methylcytidine, 5-methyluridine and other nucleotides having a modification at the 5 position, 5-(2-amino) propyl uridine, 5-halocytidine, 5-halouridine, 4-acetylcytidine, 1-methyladenosine, 2-methyladenosine, 3-methylcytidine, 6-methyluridine, 2-methylguanosine, 7-methylguanosine, 2, 2-dimethylguanosine, 5-methylaminoethyluridine, 5-methyloxyuridine, deazanucleotides (such as 7-deaza-adenosine, 6-azouridine, 6-azocytidine, or 6-azothymidine), 5-methyl-2-thiouridine, other thio bases (such as 2-thiouridine, 4-thiouridine, and 2-thiocytidine), dihydrouridine, pseudouridine, queuosine, archaeosine, naphthyl and substituted naphthyl groups, any O- and N-alkylated purines and pyrimidines (such as N6-methyladenosine, 5-methylcarbonylmethyluridine, uridine 5-oxyacetic acid, pyridine-4-one, or pyridine-2-one), phenyl and modified phenyl groups such as aminophenol or 2,4, 6-trimethoxy benzene, modified cytosines that act as G-clamp nucleotides, 8-substituted adenines and guanines, 5-substituted uracils and thymines, azapyrimidines, carboxyhydroxyalkyl nucleotides, carboxyalkylaminoalkyi nucleotides, and alkylcarbonylalkylated nucleotides. 5′-Vinylphosphonate modified nucleotides also include those nucleotides that are modified with respect to the sugar moiety, as well as 5′-vinylphosphonate modified nucleotides having sugars or analogs thereof that are not ribosyl. For example, the sugar moieties, in some cases are or are based on, mannoses, arabinoses, glucopyranoses, galactopyranoses, 4′-thioribose, and other sugars, heterocycles, or carbocycles. The term nucleotide also includes what are known in the art as universal bases. By way of example, universal bases include but are not limited to 3-nitropyrrole, 5-nitroindole, or nebularine. 
     In some embodiments, a 5′-vinylphosphonate modified nucleotide analogue further comprises a morpholino, a peptide nucleic acid (PNA), a methylphosphonate nucleotide, a thiolphosphonate nucleotide, a 2′-fluoro N3-P5′-phosphoramidite, or a 1′,5′-anhydrohexitol nucleic acid (HNA). Morpholino or phosphorodiamidate morpholino oligo (PMO) comprises synthetic molecules whose structure mimics natural nucleic acid structure but deviates from the normal sugar and phosphate structures. In some instances, the five member ribose ring is substituted with a six member morpholino ring containing four carbons, one nitrogen, and one oxygen. In some cases, the ribose monomers are linked by a phosphordiamidate group instead of a phosphate group. In such cases, the backbone alterations remove all positive and negative charges making morpholinos neutral molecules capable of crossing cellular membranes without the aid of cellular delivery agents such as those used by charged oligonucleotides. A non-limiting example of a 5′-vinylphosphonate modified morpholino oligonucleotide is illustrated below, wherein X is O or S; and B is a heterocyclic base moiety. 
     
       
         
         
             
             
         
       
     
     In some embodiments, a 5′-vinylphosphonate modified morpholino or PMO described above is a PMO comprising a positive or cationic charge. In some instances, the PMO is PMOplus (Sarepta). PMOplus refers to phosphorodiamidate morpholino oligomers comprising any number of (1-piperazino)phosphinylideneoxy, (1-(4-(omega-guanidino-alkanoyl))-piperazino)phosphinylideneoxy linkages (e.g., as such those described in PCT Publication No. WO2008/036127. In some cases, the PMO is a PMO described in U.S. Pat. No. 7,943,762. 
     In some embodiments, a morpholino or PMO described above is a PMO-X (Sarepta). In some cases, PMO-X refers to phosphorodiamidate morpholino oligomers comprising at least one linkage or at least one of the disclosed terminal modifications, such as those disclosed in PCT Publication No. WO2011/150408 and U.S. Publication No. 2012/0065169. 
     In some embodiments, a morpholino or PMO described above is a PMO as described in Table 5 of U.S. Publication No. 2014/0296321. 
     Exemplary representations of the chemical structure of 5′-vinylphosphonate modified nucleic acids are illustrated below, wherein X is O or S; B is a heterocyclic base moiety; and J is an internucleotide linkage. 
     
       
         
         
             
             
         
       
     
     In some embodiments, peptide nucleic acid (PNA) does not contain sugar ring or phosphate linkage and the bases are attached and appropriately spaced by oligoglycine-like molecules, therefore, eliminating a backbone charge. 
     
       
         
         
             
             
         
       
     
     In some embodiments, one or more modifications of the 5′-vinylphosphonate modified oligonucleotide optionally occur at the internucleotide linkage. In some instances, modified internucleotide linkage includes, but is not limited to, phosphorothioates; phosphorodithioates; methylphosphonates; 5′-alkylenephosphonates; 5′-methylphosphonate; 3′-alkylene phosphonates; borontrifluoridates; borano phosphate esters and selenophosphates of 3′-5′linkage or 2′-5′linkage; phosphotriesters; thionoalkylphosphotriesters; hydrogen phosphonate linkages; alkyl phosphonates; alkylphosphonothioates; arylphosphonothioates; phosphoroselenoates; phosphorodiselenoates; phosphinates; phosphoramidates; 3′-alkylphosphoramidates; aminoalkylphosphoramidates; thionophosphoramidates; phosphoropiperazidates; phosphoroanilothioates; phosphoroanilidates; ketones; sulfones; sulfonamides; carbonates; carbamates; methylenehydrazos; methylenedimethylhydrazos; formacetals; thioformacetals; oximes; methyleneiminos; methylenemethyliminos; thioamidates; linkages with riboacetyl groups; aminoethyl glycine; silyl or siloxane linkages; alkyl or cycloalkyl linkages with or without heteroatoms of, for example, 1 to 10 carbons that are saturated or unsaturated and/or substituted and/or contain heteroatoms; linkages with morpholino structures, amides, or polyamides wherein the bases are attached to the aza nitrogens of the backbone directly or indirectly; and combinations thereof. 
     In some instances, the modification is a methyl or thiol modification such as methylphosphonate or thiolphosphonate modification. Exemplary thiolphosphonate nucleotide (left), phosphorodithioates (center) and methylphosphonate nucleotide (right) are illustrated below. 
     
       
         
         
             
             
         
       
     
     In some instances, a 5′-vinylphosphonate modified nucleotide includes, but is not limited to, phosphoramidites illustrated as: 
     
       
         
         
             
             
         
       
     
     In some instances, the modified internucleotide linkage is a phosphorodiamidate linkage. A non-limiting example of a phosphorodiamidate linkage with a morpholino system is shown below. 
     
       
         
         
             
             
         
       
     
     In some instances, the modified internucleotide linkage is a methylphosphonate linkage. A non-limiting example of a methylphosphonate linkage is shown below. 
     
       
         
         
             
             
         
       
     
     In some instances, the modified internucleotide linkage is a amide linkage. A non-limiting example of an amide linkage is shown below. 
     
       
         
         
             
             
         
       
     
     In some instances, a 5′-vinylphosphonate modified nucleotide includes, but is not limited to, the modified nucleic acid illustrated below. 
     In some embodiments, one or more modifications comprise a modified phosphate backbone in which the modification generates a neutral or uncharged backbone. In some instances, the phosphate backbone is modified by alkylation to generate an uncharged or neutral phosphate backbone. As used herein, alkylation includes methylation, ethylation, and propylation. In some cases, an alkyl group, as used herein in the context of alkylation, refers to a linear or branched saturated hydrocarbon group containing from 1 to 6 carbon atoms. In some instances, exemplary alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, hexyl, isohexyl, 1, 1-dimethylbutyl, 2,2-dimethylbutyl, 3.3-dimethylbutyl, and 2-ethylbutyl groups. In some cases, a modified phosphate is a phosphate group as described in U.S. Pat. No. 9,481,905. 
     In some embodiments, additional modified phosphate backbones comprise methylphosphonate, ethylphosphonate, methylthiophosphonate, or methoxyphosphonate. In some cases, the modified phosphate is methylphosphonate. In some cases, the modified phosphate is ethylphosphonate. In some cases, the modified phosphate is methylthiophosphonate. In some cases, the modified phosphate is methoxyphosphonate. 
     In some embodiments, one or more modifications further optionally include modifications of the ribose moiety, phosphate backbone and the nucleoside, or modifications of the nucleotide analogues at the 3′ or the 5′ terminus. For example, the 3′ terminus optionally include a 3′ cationic group, or by inverting the nucleoside at the 3′-terminus with a 3′-3′ linkage. In another alternative, the 3′-terminus is optionally conjugated with an arninoalkyl group, e.g., a 3′ C5-aminoalkyl dT. In an additional alternative, the 3′-terminus is optionally conjugated with an abasic site, e.g., with an apurinic or apyrimidinic site. In some instances, the 5′-terminus is conjugated with an aminoalkyl group, e.g., a 5′-O-alkylamino substituent. In some cases, the 5′-terminus is conjugated with an abasic site, e.g., with an apurinic or apyrimidinic site, 
     In some embodiments, the polynucleic acid molecule comprises one or more of the artificial nucleotide analogues described herein. In some instances, the polynucleic acid molecule comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 25, or more of the artificial nucleotide analogues described herein. In some embodiments, the artificial nucleotide analogues include 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified, LNA, ENA, PNA, HNA, morpholino, methylphosphonate nucleotides, thiolphosphonate nucleotides, 2′-fluoro N3-P5′-phosphoramidites, or a combination thereof. In some instances, the polynucleic acid molecule comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 25, or more of the artificial nucleotide analogues selected from 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified, LNA, ENA, PNA, HNA, morpholino, methylphosphonate nucleotides, thiolphosphonate nucleotides, 2′-fluoro N3-P5′-phosphoramidites, or a combination thereof. In some instances, the polynucleic acid molecule comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 25, or more of 2′-O-methyl modified nucleotides. In some instances, the polynucleic acid molecule comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 25, or more of 2′-O-methoxyethyl (2′-O-MOE) modified nucleotides. In some instances, the polynucleic acid molecule comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 20, 25, or more of thiolphosphonate nucleotides. 
     In some embodiments, the polynucleic acid molecule comprises a plurality of phosphorodiamidate morpholino oligomers or a plurality of peptide nucleic acid-modified non-natural nucleotides, and optionally comprises at least one inverted abasic moiety. In some instances, the polynucleic acid molecule comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorodiamidate morpholino oligomer-modified non-natural nucleotides. In some instances, the polynucleic acid molecule comprises 100% phosphorodiamidate morpholino oligomer-modified non-natural nucleotides. 
     In some instances, the polynucleic acid molecule comprises at least 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more peptide nucleic acid-modified non-natural nucleotides. In some instances, the polynucleic acid molecule comprises 100% peptide nucleic acid-modified non-natural nucleotides. 
     In some embodiments, the polynucleic acid molecule comprises one or more nucleotide analogs in which each nucleotide analog is in a stereochemically isomeric form. In such instance, the polynucleic acid molecule is a chiral molecule. In some cases, the nucleotide analog comprises a backbone stereochemistry. In additional cases, the nucleotide analog comprises a chiral analog as described in U.S. Pat. Nos. 9,982,257, 9,695,211, or 9,605,019. 
     In some instances, the polynucleic acid molecule comprises at least one of: from about 5% to about 100% modification, from about 10% to about 100% modification, from about 20% to about 100% modification, from about 30% to about 100% modification, from about 40% to about 100% modification, from about 50% to about 100% modification, from about 60% to about 100% modification, from about 70% to about 100% modification, from about 80% to about 100% modification, and from about 90% to about 100% modification. 
     In some cases, the polynucleic acid molecule comprises at least one of: from about 10% to about 90% modification, from about 20% to about 90% modification, from about 30% to about 90% modification, from about 40% to about 90% modification, from about 50% to about 90% modification, from about 60% to about 90% modification, from about 70% to about 90% modification, and from about 80% to about 100% modification. 
     In some cases, the polynucleic acid molecule comprises at least one of: from about 10% to about 80% modification, from about 20% to about 80% modification, from about 30% to about 80% modification, from about 40% to about 80% modification, from about 50% to about 80% modification, from about 60% to about 80% modification, and from about 70% to about 80% modification. 
     In some instances, the polynucleic acid molecule comprises at least one of: from about 10% to about 70% modification, from about 20% to about 70% modification, from about 30% to about 70% modification, from about 40% to about 70% modification, from about 50% to about 70% modification, and from about 60% to about 70% modification. 
     In some instances, the polynucleic acid molecule comprises at least one of: from about 10% to about 60% modification, from about 20% to about 60% modification, from about 30% to about 60% modification, from about 40% to about 60% modification, and from about 50% to about 60% modification. 
     In some cases, the polynucleic acid molecule comprises at least one of: from about 10% to about 50% modification, from about 20% to about 50% modification, from about 30% to about 50% modification, and from about 40% to about 50% modification. 
     In some cases, the polynucleic acid molecule comprises at least one of: from about 10% to about 40% modification, from about 20% to about 40% modification, and from about 30% to about 40% modification. 
     In some cases, the polynucleic acid molecule comprises at least one of: from about 10% to about 30% modification, and from about 20% to about 30% modification. 
     In some cases, the polynucleic acid molecule comprises from about 10% to about 20% modification. 
     In some cases, the polynucleic acid molecule comprises from about 15% to about 90%, from about 20% to about 80%, from about 30% to about 70%, or from about 40% to about 60% modifications. 
     In additional cases, the polynucleic acid molecule comprises at least about 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% modification. 
     In some embodiments, the polynucleic acid molecule comprises at least about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22 or more modifications. 
     In some instances, the polynucleic acid molecule comprises at least about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22 or more modified nucleotides. 
     In some instances, from about 5 to about 100% of the polynucleic acid molecule comprise the artificial nucleotide analogues described herein. In some instances, about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 100% of the polynucleic acid molecule comprise the artificial nucleotide analogues described herein. In some instances, about 5% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 10% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 15% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 20% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 25% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 30% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 35% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 40% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 45% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 50% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 55% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 60% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 65% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 70% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 75% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 80% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 85% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 90% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 95% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 96% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 97% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 98% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 99% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some instances, about 100% of a polynucleic acid molecule comprises the artificial nucleotide analogues described herein. In some embodiments, the artificial nucleotide analogues include 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified, LNA, ENA, PNA, HNA, morpholino, methylphosphonate nucleotides, thiolphosphonate nucleotides, 2′-fluoro N3-P5′-phosphoramidites, or a combination thereof. 
     In some embodiments, the polynucleic acid molecule comprises from about 1 to about 25 modifications in which the modification comprises an artificial nucleotide analogues described herein. In some embodiments, a polynucleic acid molecule comprises about 1 modification in which the modification comprises an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 2 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 3 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 4 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 5 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 6 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 7 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 8 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 9 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 10 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 11 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 12 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 13 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 14 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 15 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 16 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 17 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 18 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 19 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 20 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 21 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 22 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 23 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 24 modifications in which the modifications comprise an artificial nucleotide analogue described herein. In some embodiments, a polynucleic acid molecule comprises about 25 modifications in which the modifications comprise an artificial nucleotide analogue described herein. 
     In some embodiments, a polynucleic acid molecule is assembled from two separate polynucleotides wherein one polynucleotide comprises the sense strand and the second polynucleotide comprises the antisense strand of the polynucleic acid molecule. In other embodiments, the sense strand is connected to the antisense strand via a linker molecule, which in some instances is a polynucleotide linker or a non-nucleotide linker. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and antisense strand, wherein pyrimidine nucleotides in the sense strand comprises 2′-O-methylpyrimidine nucleotides and purine nucleotides in the sense strand comprise 2′-deoxy purine nucleotides. In some embodiments, a polynucleic acid molecule comprises a sense strand and antisense strand, wherein pyrimidine nucleotides present in the sense strand comprise 2′-deoxy-2′-fluoro pyrimidine nucleotides and wherein purine nucleotides present in the sense strand comprise 2′-deoxy purine nucleotides. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and antisense strand, wherein the pyrimidine nucleotides when present in said antisense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and the purine nucleotides when present in said antisense strand are 2′-O-methyl purine nucleotides. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and antisense strand, wherein the pyrimidine nucleotides when present in said antisense strand are 2′-deoxy-2′-fluoro pyrimidine nucleotides and wherein the purine nucleotides when present in said antisense strand comprise 2′-deoxy-purine nucleotides. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and antisense strand, wherein the sense strand includes a terminal cap moiety at the 5′-end, the 3′-end, or both of the 5′ and 3′ ends of the sense strand. In other embodiments, the terminal cap moiety is an inverted deoxy abasic moiety. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and an antisense strand, wherein the antisense strand comprises a phosphate backbone modification at the 3′ end of the antisense strand. In some instances, the phosphate backbone modification is a phosphorothioate. In some cases, the passenger strand comprises more phosphorothioate modifications than the guide strand. In other cases, the guide strand comprises more phosphorothioate modifications than the passenger strand. In additional cases, the passenger strand comprises about 2, 3, 4, 5, 6, 7, 8, 9, 10, or more phosphorothioate modifications. In additional cases, the guide strand comprises about 2, 3, 4, 5, 6, 7, 8, 9, 10, or more phosphorothioate modifications. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and an antisense strand, wherein the antisense strand comprises a glyceryl modification at the 3′ end of the antisense strand. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and an antisense strand, in which the sense strand comprises one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, and/or about one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and in which the antisense strand comprises about 1 to about 10 or more, specifically about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In other embodiments, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, pyrimidine nucleotides of the sense and/or antisense strand are chemically-modified with 2′-deoxy, 2′-O-methyl and/or 2′-deoxy-2′-fluoro nucleotides, with or without one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and an antisense strand, in which the sense strand comprises about 1 to about 25, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and in which the antisense strand comprises about 1 to about 25 or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In other embodiments, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more, pyrimidine nucleotides of the sense and/or antisense strand are chemically-modified with 2′-deoxy, 2′-O-methyl and/or 2′-deoxy-2′-fluoro nucleotides, with or without about 1 to about 25 or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and an antisense strand, in which the antisense strand comprises one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorothioate internucleotide linkages, and/or about one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 10 or more, specifically about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In other embodiments, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more pyrimidine nucleotides of the sense and/or antisense strand are chemically-modified with 2′-deoxy, 2′-O-methyl and/or 2′-deoxy-2′-fluoro nucleotides, with or without one or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends, being present in the same or different strand. 
     In some embodiments, a polynucleic acid molecule comprises a sense strand and an antisense strand, in which the antisense strand comprises about 1 to about 25 or more, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the sense strand; and wherein the antisense strand comprises about 1 to about 25 or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more phosphorothioate internucleotide linkages, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) 2′-deoxy, 2′-O-methyl, 2′-deoxy-2′-fluoro, and/or one or more (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more) universal base modified nucleotides, and optionally a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of the antisense strand. In other embodiments, one or more, for example about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more pyrimidine nucleotides of the sense and/or antisense strand are chemically-modified with 2′-deoxy, 2′-O-methyl and/or 2′-deoxy-2′-fluoro nucleotides, with or without about 1 to about 5, for example about 1, 2, 3, 4, 5 or more phosphorothioate internucleotide linkages and/or a terminal cap molecule at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends, being present in the same or different strand. 
     In some embodiments, a polynucleic acid molecule is a duplex polynucleic acid molecule with one or more of the following properties: a greater hepatocyte stability, reduced overall charge, reduced hepatocyte uptake, or extended pharmacokinetics. In some embodiments, the duplex polynucleic acid molecule comprises a passenger strand (e.g., a sense strand) and a guide strand (e.g., an antisense strand) comprising a plurality of modifications. 
     In some embodiments, the duplex polynucleic acid molecule comprises a guide strand (e.g., an antisense strand) with one or more of the modification described above, and a passenger strand (e.g., a sense strand) with a plurality of phosphorodiamidate morpholino oligomers or a plurality of peptide nucleic acid-modified non-natural nucleotides. 
     In some embodiments, a polynucleic acid molecule described herein is a chemically-modified short interfering nucleic acid molecule having about 1 to about 25, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more phosphorothioate internucleotide linkages in each strand of the polynucleic acid molecule. 
     In another embodiment, a polynucleic acid molecule described herein comprises 2′-5′ internucleotide linkages. In some instances, the 2′-5′ internucleotide linkage(s) is at the 3′-end, the 5′-end, or both of the 3′- and 5′-ends of one or both sequence strands. In addition instances, the 2′-5′ internucleotide linkage(s) is present at various other positions within one or both sequence strands, for example, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more including every internucleotide linkage of a pyrimidine nucleotide in one or both strands of the polynucleic acid molecule comprise a 2′-5′ internucleotide linkage, or about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more including every internucleotide linkage of a purine nucleotide in one or both strands of the polynucleic acid molecule comprise a 2′-5′ internucleotide linkage. 
     In some embodiments, a polynucleic acid molecule is a single stranded polynucleic acid molecule that mediates RNAi activity in a cell or reconstituted in vitro system, wherein the polynucleic acid molecule comprises a single stranded polynucleotide having complementarity to a target nucleic acid sequence, and wherein one or more pyrimidine nucleotides present in the polynucleic acid are 2′-deoxy-2′-fluoro pyrimidine nucleotides (e.g., wherein all pyrimidine nucleotides are 2′-deoxy-2′-fluoro pyrimidine nucleotides or alternately a plurality of pyrimidine nucleotides are 2′-deoxy-2′-fluoro pyrimidine nucleotides), and wherein any purine nucleotides present in the polynucleic acid are 2′-deoxy purine nucleotides (e.g., wherein all purine nucleotides are 2′-deoxy purine nucleotides or alternately a plurality of purine nucleotides are 2′-deoxy purine nucleotides), and a terminal cap modification, that is optionally present at the 3′-end, the 5′-end, or both of the 3′ and 5′-ends of the antisense sequence, the polynucleic acid molecule optionally further comprising about 1 to about 4 (e.g., about 1, 2, 3, or 4) terminal 2′-deoxynucleotides at the 3′-end of the polynucleic acid molecule, wherein the terminal nucleotides further comprise one or more (e.g., 1, 2, 3, or 4) phosphorothioate internucleotide linkages, and wherein the polynucleic acid molecule optionally further comprises a terminal phosphate group, such as a 5′-terminal phosphate group. 
     In some cases, one or more of the artificial nucleotide analogues described herein are resistant toward nucleases such as for example ribonuclease such as RNase H, deoxyribunuclease such as DNase, or exonuclease such as 5′-3′ exonuclease and 3′-5′ exonuclease when compared to natural polynucleic acid molecules. In some instances, artificial nucleotide analogues comprising 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified, LNA, ENA, PNA, HNA, morpholino, methylphosphonate nucleotides, thiolphosphonate nucleotides, 2′-fluoro N3-P5′-phosphoramidites, or combinations thereof are resistant toward nucleases such as for example ribonuclease such as RNase H, deoxyribunuclease such as DNase, or exonuclease such as 5′-3′ exonuclease and 3′-5′ exonuclease. In some instances, 2′-O-methyl modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, 2′O-methoxyethyl (2′-O-MOE) modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, 2′-O-aminopropyl modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, 2′-deoxy modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, T-deoxy-2′-fluoro modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, 2′-O-aminopropyl (2′-O-AP) modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, 2′-O-dimethylaminoethyl (2′-O-DMAOE) modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, 2′-O-dimethylaminopropyl (2′-O-DMAP) modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE) modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, 2′-O—N-methylacetamido (2′-O-NMA) modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, LNA modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, ENA modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, HNA modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, morpholinos is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, PNA modified polynucleic acid molecule is resistant to nucleases (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, methylphosphonate nucleotides modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, thiolphosphonate nucleotides modified polynucleic acid molecule is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, polynucleic acid molecule comprising 2′-fluoro N3-P5′-phosphoramidites is nuclease resistance (e.g., RNase H, DNase, 5′-3′ exonuclease or 3′-5′ exonuclease resistance). In some instances, the 5′ conjugates described herein inhibit 5′-3′ exonucleolytic cleavage. In some instances, the 3′ conjugates described herein inhibit 3′-5′ exonucleolytic cleavage. 
     In some embodiments, one or more of the artificial nucleotide analogues described herein have increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. The one or more of the artificial nucleotide analogues comprising 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modified, LNA, ENA, PNA, HNA, morpholino, methylphosphonate nucleotides, thiolphosphonate nucleotides, or 2′-fluoro N3-P5′-phosphoramidites have increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, 2′-O-methyl modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, 2′-O-methoxyethyl (2′-O-MOE) modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, 2′-O-aminopropyl modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, 2′-deoxy modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, T-deoxy-2′-fluoro modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, 2′-O-aminopropyl (2′-O-AP) modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, 2′-O-dimethylaminoethyl (2′-O-DMAOE) modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, 2′-O-dimethylaminopropyl (2′-O-DMAP) modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE) modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, 2′-O—N-methylacetamido (2′-O-NMA) modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, LNA modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, ENA modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, PNA modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, HNA modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, morpholino modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, methylphosphonate nucleotides modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, thiolphosphonate nucleotides modified polynucleic acid molecule has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some instances, polynucleic acid molecule comprising 2′-fluoro N3-P5′-phosphoramidites has increased binding affinity toward their mRNA target relative to an equivalent natural polynucleic acid molecule. In some cases, the increased affinity is illustrated with a lower Kd, a higher melt temperature (Tm), or a combination thereof. 
     In some embodiments, a polynucleic acid molecule described herein is a chirally pure (or stereo pure) polynucleic acid molecule, or a polynucleic acid molecule comprising a single enantiomer. In some instances, the polynucleic acid molecule comprises L-nucleotide. In some instances, the polynucleic acid molecule comprises D-nucleotides. In some instance, a polynucleic acid molecule composition comprises less than 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, or less of its mirror enantiomer. In some cases, a polynucleic acid molecule composition comprises less than 30%, 25%, 20%, 15%, 10%, 5%, 4%, 3%, 2%, 1%, or less of a racemic mixture. In some instances, the polynucleic acid molecule is a polynucleic acid molecule described in: U.S. Patent Publication Nos: 2014/194610 and 2015/211006; and PCT Publication No.: WO2015107425. 
     In some embodiments, a polynucleic acid molecule described herein is further modified to include an aptamer conjugating moiety. In some instances, the aptamer conjugating moiety is a DNA aptamer conjugating moiety. In some instances, the aptamer conjugating moiety is Alphamer (Centauri Therapeutics), which comprises an aptamer portion that recognizes a specific cell-surface target and a portion that presents a specific epitopes for attaching to circulating antibodies. In some instance, a polynucleic acid molecule described herein is further modified to include an aptamer conjugating moiety as described in: U.S. Pat. Nos. 8,604,184, 8,591,910, and 7,850,975. 
     In additional embodiments, a polynucleic acid molecule described herein is modified to increase its stability. In some embodiment, the polynucleic acid molecule is RNA (e.g., siRNA). In some instances, the polynucleic acid molecule is modified by one or more of the modifications described above to increase its stability. In some cases, the polynucleic acid molecule is modified at the 2′ hydroxyl position, such as by 2′-O-methyl, 2′-O-methoxyethyl (2′-O-MOE), 2′-O-aminopropyl, 2′-deoxy, T-deoxy-2′-fluoro, 2′-O-aminopropyl (2′-O-AP), 2′-O-dimethylaminoethyl (2′-O-DMAOE), 2′-O-dimethylaminopropyl (2′-O-DMAP), T-O-dimethylaminoethyloxyethyl (2′-O-DMAEOE), or 2′-O—N-methylacetamido (2′-O-NMA) modification or by a locked or bridged ribose conformation (e.g., LNA or ENA). In some cases, the polynucleic acid molecule is modified by 2′-O-methyl and/or 2′-O-methoxyethyl ribose. In some cases, the polynucleic acid molecule also includes morpholinos, PNAs, HNA, methylphosphonate nucleotides, thiolphosphonate nucleotides, and/or 2′-fluoro N3-P5′-phosphoramidites to increase its stability. In some instances, the polynucleic acid molecule is a chirally pure (or stereo pure) polynucleic acid molecule. In some instances, the chirally pure (or stereo pure) polynucleic acid molecule is modified to increase its stability. Suitable modifications to the RNA to increase stability for delivery will be apparent to the skilled person. 
     In some embodiments, a polynucleic acid molecule describe herein has RNAi activity that modulates expression of RNA encoded by a gene involved in muscular dystrophy such as, but not limited to, DMD, DUX4, DYSF, EMD, or LMNA. In some instances, a polynucleic acid molecule describe herein is a double-stranded siRNA molecule that down-regulates expression of at least one of DMD, DUX4, DYSF, EMD, or LMNA, wherein one of the strands of the double-stranded siRNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence of at least one of DMD, DUX4, DYSF, EMD, or LMNA or RNA encoded by at least one of DMD, DUX4, DYSF, EMD, or LMNA or a portion thereof, and wherein the second strand of the double-stranded siRNA molecule comprises a nucleotide sequence substantially similar to the nucleotide sequence of at least one of DMD, DUX4, DYSF, EMD, or LMNA or RNA encoded by at least one of DMD, DUX4, DYSF, EMD, or LMNA or a portion thereof. In some cases, a polynucleic acid molecule describe herein is a double-stranded siRNA molecule that down-regulates expression of at least one of DMD, DUX4, DYSF, EMD, or LMNA, wherein each strand of the siRNA molecule comprises about 15 to 25, 18 to 24, or 19 to about 23 nucleotides, and wherein each strand comprises at least about 14, 17, or 19 nucleotides that are complementary to the nucleotides of the other strand. In some cases, a polynucleic acid molecule describe herein is a double-stranded siRNA molecule that down-regulates expression of at least one of DMD, DUX4, DYSF, EMD, or LMNA, wherein each strand of the siRNA molecule comprises about 19 to about 23 nucleotides, and wherein each strand comprises at least about 19 nucleotides that are complementary to the nucleotides of the other strand. In some instances, the RNAi activity occurs within a cell. In other instances, the RNAi activity occurs in a reconstituted in vitro system. 
     In some embodiments, a polynucleic acid molecule describe herein has RNAi activity that modulates expression of RNA encoded by the DMD gene. In some instances, a polynucleic acid molecule describe herein is a single-stranded siRNA molecule that down-regulates expression of DMD, wherein the single-stranded siRNA molecule comprises a nucleotide sequence that is complementary to a nucleotide sequence of DMD or RNA encoded by DMD or a portion thereof. In some cases, a polynucleic acid molecule describe herein is a single-stranded siRNA molecule that down-regulates expression of DMD, wherein the siRNA molecule comprises about 15 to 25, 18 to 24, or 19 to about 23 nucleotides. In some cases, a polynucleic acid molecule describe herein is a single-stranded siRNA molecule that down-regulates expression of DMD, wherein the siRNA molecule comprises about 19 to about 23 nucleotides. In some instances, the RNAi activity occurs within a cell. In other instances, the RNAi activity occurs in a reconstituted in vitro system. 
     In some instances, the polynucleic acid molecule is a double-stranded polynucleotide molecule comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. In some instances, the polynucleic acid molecule is assembled from two separate polynucleotides, where one strand is the sense strand and the other is the antisense strand, wherein the antisense and sense strands are self-complementary (e.g., each strand comprises nucleotide sequence that is complementary to nucleotide sequence in the other strand; such as where the antisense strand and sense strand form a duplex or double stranded structure, for example wherein the double stranded region is about 19, 20, 21, 22, 23, or more base pairs); the antisense strand comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense strand comprises nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. Alternatively, the polynucleic acid molecule is assembled from a single oligonucleotide, where the self-complementary sense and antisense regions of the polynucleic acid molecule are linked by means of a nucleic acid based or non-nucleic acid-based linker(s). 
     In some cases, the polynucleic acid molecule is a polynucleotide with a duplex, asymmetric duplex, hairpin or asymmetric hairpin secondary structure, having self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a separate target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof. In other cases, the polynucleic acid molecule is a circular single-stranded polynucleotide having two or more loop structures and a stem comprising self-complementary sense and antisense regions, wherein the antisense region comprises nucleotide sequence that is complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof and the sense region having nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof, and wherein the circular polynucleotide is processed either in vivo or in vitro to generate an active polynucleic acid molecule capable of mediating RNAi. In additional cases, the polynucleic acid molecule also comprises a single stranded polynucleotide having nucleotide sequence complementary to nucleotide sequence in a target nucleic acid molecule or a portion thereof (for example, where such polynucleic acid molecule does not require the presence within the polynucleic acid molecule of nucleotide sequence corresponding to the target nucleic acid sequence or a portion thereof), wherein the single stranded polynucleotide further comprises a terminal phosphate group, such as a 5′-phosphate (see for example Martinez et al., 2002 , Cell.,  110, 563-574 and Schwarz et al., 2002 , Molecular Cell,  10, 537-568), or 5′,3′-diphosphate. 
     In some instances, an asymmetric is a linear polynucleic acid molecule comprising an antisense region, a loop portion that comprises nucleotides or non-nucleotides, and a sense region that comprises fewer nucleotides than the antisense region to the extent that the sense region has enough complimentary nucleotides to base pair with the antisense region and form a duplex with loop. For example, an asymmetric hairpin polynucleic acid molecule comprises an antisense region having length sufficient to mediate RNAi in a cell or in vitro system (e.g. about 19 to about 22 nucleotides) and a loop region comprising about 4 to about 8 nucleotides, and a sense region having about 3 to about 18 nucleotides that are complementary to the antisense region. In some cases, the asymmetric hairpin polynucleic acid molecule also comprises a 5′-terminal phosphate group that is chemically modified. In additional cases, the loop portion of the asymmetric hairpin polynucleic acid molecule comprises nucleotides, non-nucleotides, linker molecules, or conjugate molecules. 
     In some embodiments, an asymmetric duplex is a polynucleic acid molecule having two separate strands comprising a sense region and an antisense region, wherein the sense region comprises fewer nucleotides than the antisense region to the extent that the sense region has enough complimentary nucleotides to base pair with the antisense region and form a duplex. For example, an asymmetric duplex polynucleic acid molecule comprises an antisense region having length sufficient to mediate RNAi in a cell or in vitro system (e.g. about 19 to about 22 nucleotides) and a sense region having about 3 to about 18 nucleotides that are complementary to the antisense region. 
     In some cases, an universal base refers to nucleotide base analogs that form base pairs with each of the natural DNA/RNA bases with little discrimination between them. Non-limiting examples of universal bases include C-phenyl, C-naphthyl and other aromatic derivatives, inosine, azole carboxamides, and nitroazole derivatives such as 3-nitropyrrole, 4-nitroindole, 5-nitroindole, and 6-nitroindole as known in the art (see for example Loakes, 2001, Nucleic Acids Research, 29, 2437-2447). 
     Polynucleic Acid Molecule Synthesis 
     In some embodiments, a polynucleic acid molecule described herein is constructed using chemical synthesis and/or enzymatic ligation reactions using procedures known in the art. For example, a polynucleic acid molecule is chemically synthesized using naturally occurring nucleotides or variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the polynucleic acid molecule and target nucleic acids. Exemplary methods include those described in: U.S. Pat. Nos. 5,142,047; 5,185,444; 5,889,136; 6,008,400; and 6,111,086; PCT Publication No. WO2009099942; or European Publication No. 1579015. Additional exemplary methods include those described in: Griffey et al., “2′-O-aminopropyl ribonucleotides: a zwitterionic modification that enhances the exonuclease resistance and biological activity of antisense oligonucleotides,”  J. Med. Chem.  39(26):5100-5109 (1997)); Obika, et al. “Synthesis of 2′-O,4′-C-methyleneuridine and -cytidine. Novel bicyclic nucleosides having a fixed C3, -endo sugar puckering”.  Tetrahedron Letters  38 (50): 8735 (1997); Koizumi, M. “ENA oligonucleotides as therapeutics”.  Current opinion in molecular therapeutics  8 (2): 144-149 (2006); and Abramova et al., “Novel oligonucleotide analogues based on morpholino nucleoside subunits-antisense technologies: new chemical possibilities,”  Indian Journal of Chemistry  48B:1721-1726 (2009). Alternatively, the polynucleic acid molecule is produced biologically using an expression vector into which a polynucleic acid molecule has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted polynucleic acid molecule will be of an antisense orientation to a target polynucleic acid molecule of interest). 
     In some embodiments, a polynucleic acid molecule is synthesized via a tandem synthesis methodology, wherein both strands are synthesized as a single contiguous oligonucleotide fragment or strand separated by a cleavable linker which is subsequently cleaved to provide separate fragments or strands that hybridize and permit purification of the duplex. 
     In some instances, a polynucleic acid molecule is also assembled from two distinct nucleic acid strands or fragments wherein one fragment includes the sense region and the second fragment includes the antisense region of the molecule. 
     Additional modification methods for incorporating, for example, sugar, base and phosphate modifications include: Eckstein et al., International Publication PCT No. WO 92/07065; Perrault et al.  Nature,  1990, 344, 565-568; Pieken et al.  Science,  1991, 253, 314-317; Usman and Cedergren,  Trends in Biochem. Sci.,  1992, 17, 334-339; Usman et al. International Publication PCT No. WO 93/15187; Sproat, U.S. Pat. No. 5,334,711 and Beigelman et al., 1995 , J Biol. Chem.,  270, 25702; Beigelman et al., International PCT publication No. WO 97/26270; Beigelman et al., U.S. Pat. No. 5,716,824; Usman et al., U.S. Pat. No. 5,627,053; Woolf et al., International PCT Publication No. WO 98/13526; Thompson et al., U.S. Ser. No. 60/082,404 which was filed on Apr. 20, 1998; Karpeisky et al., 1998 , Tetrahedron Lett.,  39, 1131; Earnshaw and Gait, 1998 , Biopolymers  ( Nucleic Acid Sciences ), 48, 39-55; Verma and Eckstein, 1998 , Annu. Rev. Biochem.,  67, 99-134; and Burlina et al., 1997 , Bioorg. Med. Chem.,  5, 1999-2010. Such publications describe general methods and strategies to determine the location of incorporation of sugar, base and/or phosphate modifications and the like into nucleic acid molecules without modulating catalysis. 
     In some instances, while chemical modification of the polynucleic acid molecule internucleotide linkages with phosphorothioate, phosphorodithioate, and/or 5′-methylphosphonate linkages improves stability, excessive modifications sometimes cause toxicity or decreased activity. Therefore, when designing nucleic acid molecules, the amount of these internucleotide linkages in some cases is minimized. In such cases, the reduction in the concentration of these linkages lowers toxicity, increases efficacy and higher specificity of these molecules. 
     Nucleic Acid-Polypeptide Conjugate 
     In some embodiments, a polynucleic acid molecule is further conjugated to a polypeptide A for delivery to a site of interest. In some cases, a polynucleic acid molecule is conjugated to a polypeptide A and optionally a polymeric moiety. 
     In some instances, at least one polypeptide A is conjugated to at least one B. In some instances, the at least one polypeptide A is conjugated to the at least one B to form an A-B conjugate. In some embodiments, at least one A is conjugated to the 5′ terminus of B, the 3′ terminus of B, an internal site on B, or in any combinations thereof. In some instances, the at least one polypeptide A is conjugated to at least two B. In some instances, the at least one polypeptide A is conjugated to at least 2, 3, 4, 5, 6, 7, 8, or more B. 
     In some embodiments, at least one polypeptide A is conjugated at one terminus of at least one B while at least one C is conjugated at the opposite terminus of the at least one B to form an A-B-C conjugate. In some instances, at least one polypeptide A is conjugated at one terminus of the at least one B while at least one of C is conjugated at an internal site on the at least one B. In some instances, at least one polypeptide A is conjugated directly to the at least one C. In some instances, the at least one B is conjugated indirectly to the at least one polypeptide A via the at least one C to form an A-C—B conjugate. 
     In some instances, at least one B and/or at least one C, and optionally at least one D are conjugated to at least one polypeptide A. In some instances, the at least one B is conjugated at a terminus (e.g., a 5′ terminus or a 3′ terminus) to the at least one polypeptide A or are conjugated via an internal site to the at least one polypeptide A. In some cases, the at least one C is conjugated either directly to the at least one polypeptide A or indirectly via the at least one B. If indirectly via the at least one B, the at least one C is conjugated either at the same terminus as the at least one polypeptide A on B, at opposing terminus from the at least one polypeptide A, or independently at an internal site. In some instances, at least one additional polypeptide A is further conjugated to the at least one polypeptide A, to B, or to C. In additional instances, the at least one D is optionally conjugated either directly or indirectly to the at least one polypeptide A, to the at least one B, or to the at least one C. If directly to the at least one polypeptide A, the at least one D is also optionally conjugated to the at least one B to form an A-D-B conjugate or is optionally conjugated to the at least one B and the at least one C to form an A-D-B-C conjugate. In some instances, the at least one D is directly conjugated to the at least one polypeptide A and indirectly to the at least one B and the at least one C to form a D-A-B-C conjugate. If indirectly to the at least one polypeptide A, the at least one D is also optionally conjugated to the at least one B to form an A-B-D conjugate or is optionally conjugated to the at least one B and the at least one C to form an A-B-D-C conjugate. In some instances, at least one additional D is further conjugated to the at least one polypeptide A, to B, or to C. 
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     In some embodiments, a polynucleic acid molecule conjugate comprises a construct as illustrated: 
     
       
         
         
             
             
         
       
     
     The 
     
       
         
         
             
             
         
       
     
     as illustrated above is for representation purposes only and encompasses a humanized antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab′, divalent Fab2, single-chain variable fragment (scFv), diabody, minibody, nanobody, single-domain antibody (sdAb), or camelid antibody or binding fragment thereof. 
     Binding Moiety 
     In some embodiments, the binding moiety A is a polypeptide. In some instances, the polypeptide is an antibody or its fragment thereof. In some cases, the fragment is a binding fragment. In some instances, the antibody or binding fragment thereof comprises a humanized antibody or binding fragment thereof, murine antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab′, divalent Fab 2 , F(ab)′ 3  fragments, single-chain variable fragment (scFv), bis-scFv, (scFv) 2 , diabody, minibody, nanobody, triabody, tetrabody, disulfide stabilized Fv protein (dsFv), single-domain antibody (sdAb), Ig NAR, camelid antibody or binding fragment thereof, bispecific antibody or biding fragment thereof, or a chemically modified derivative thereof. 
     In some instances, A is an antibody or binding fragment thereof. In some instances, A is a humanized antibody or binding fragment thereof, murine antibody or binding fragment thereof, chimeric antibody or binding fragment thereof, monoclonal antibody or binding fragment thereof, monovalent Fab′, divalent Fab 2 , F(ab)′ 3  fragments, single-chain variable fragment (scFv), bis-scFv, (scFv) 2 , diabody, minibody, nanobody, triabody, tetrabody, disulfide stabilized Fv protein (“dsFv”), single-domain antibody (sdAb), Ig NAR, camelid antibody or binding fragment thereof, bispecific antibody or biding fragment thereof, or a chemically modified derivative thereof. In some instances, A is a humanized antibody or binding fragment thereof. In some instances, A is a murine antibody or binding fragment thereof. In some instances, A is a chimeric antibody or binding fragment thereof. In some instances, A is a monoclonal antibody or binding fragment thereof. In some instances, A is a monovalent Fab′. In some instances, A is a divalent Fab 2 . In some instances, A is a single-chain variable fragment (scFv). 
     In some embodiments, the binding moiety A is a bispecific antibody or binding fragment thereof. In some instances, the bispecific antibody is a trifunctional antibody or a bispecific mini-antibody. In some cases, the bispecific antibody is a trifunctional antibody. In some instances, the trifunctional antibody is a full length monoclonal antibody comprising binding sites for two different antigens. 
     In some cases, the bispecific antibody is a bispecific mini-antibody. In some instances, the bispecific mini-antibody comprises divalent Fab 2 , F(ab)′ 3  fragments, bis-scFv, (scFv) 2 , diabody, minibody, triabody, tetrabody or a bi-specific T-cell engager (BiTE). In some embodiments, the bi-specific T-cell engager is a fusion protein that contains two single-chain variable fragments (scFvs) in which the two scFvs target epitopes of two different antigens. 
     In some embodiments, the binding moiety A is a bispecific mini-antibody. In some instances, A is a bispecific Fab 2 . In some instances, A is a bispecific F(ab)′ 3  fragment. In some cases, A is a bispecific bis-scFv. In some cases, A is a bispecific (scFv) 2 . In some embodiments, A is a bispecific diabody. In some embodiments, A is a bispecific minibody. In some embodiments, A is a bispecific triabody. In other embodiments, A is a bispecific tetrabody. In other embodiments, A is a bi-specific T-cell engager (BiTE). 
     In some embodiments, the binding moiety A is a trispecific antibody. In some instances, the trispecific antibody comprises F(ab)′ 3  fragments or a triabody. In some instances, A is a trispecific F(ab)′ 3  fragment. In some cases, A is a triabody. In some embodiments, A is a trispecific antibody as described in Dimas, et al., “Development of a trispecific antibody designed to simultaneously and efficiently target three different antigens on tumor cells,”  Mol. Pharmaceutics,  12(9): 3490-3501 (2015). 
     In some embodiments, the binding moiety A is an antibody or binding fragment thereof that recognizes a cell surface protein. In some instances, the binding moiety A is an antibody or binding fragment thereof that recognizes a cell surface protein on a muscle cell. Exemplary cell surface proteins recognized by an antibody or binding fragment thereof include, but are not limited to, Sca-1, CD34, Myo-D, myogenin, MRF4, NCAM, CD43, and CD95 (Fas). 
     In some instances, the cell surface protein comprises clusters of differentiation (CD) cell surface markers. Exemplary CD cell surface markers include, but are not limited to, CD1, CD2, CD3, CD4, CD5, CD6, CD7, CD8, CD9, CD10, CD11a, CD11b, CD11c, CD11d, CDw12, CD13, CD14, CD15, CD15s, CD16, CDw17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD43, CD44, CD45, CD45RO, CD45RA, CD45RB, CD46, CD47, CD48, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CDw60, CD61, CD62E, CD62L (L-selectin), CD62P, CD63, CD64, CD65, CD66a, CD66b, CD66c, CD66d, CD66e, CD79 (e.g., CD79a, CD79b), CD90, CD95 (Fas), CD103, CD104, CD125 (IL5RA), CD134 (OX40), CD137 (4-1BB), CD152 (CTLA-4), CD221, CD274, CD279 (PD-1), CD319 (SLAMF7), CD326 (EpCAM), and the like. 
     In some instances, the binding moiety A is an antibody or binding fragment thereof that recognizes a CD cell surface marker. In some instances, the binding moiety A is an antibody or binding fragment thereof that recognizes CD1, CD2, CD3, CD4, CD5, CD6, CD7, CD8, CD9, CD10, CD11a, CD11b, CD11c, CD11d, CDw12, CD13, CD14, CD15, CD15s, CD16, CDw17, CD18, CD19, CD20, CD21, CD22, CD23, CD24, CD25, CD26, CD27, CD28, CD29, CD30, CD31, CD32, CD33, CD34, CD35, CD36, CD37, CD38, CD39, CD40, CD41, CD42, CD43, CD44, CD45, CD45RO, CD45RA, CD45RB, CD46, CD47, CD48, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD50, CD51, CD52, CD53, CD54, CD55, CD56, CD57, CD58, CD59, CDw60, CD61, CD62E, CD62L (L-selectin), CD62P, CD63, CD64, CD65, CD66a, CD66b, CD66c, CD66d, CD66e, CD79 (e.g., CD79a, CD79b), CD90, CD95 (Fas), CD103, CD104, CD125 (IL5RA), CD134 (OX40), CD137 (4-1BB), CD152 (CTLA-4), CD221, CD274, CD279 (PD-1), CD319 (SLAMF7), CD326 (EpCAM), or a combination thereof. 
     In some embodiments, the binding moiety A is an anti-myosin antibody, an anti-transferrin antibody, and an antibody that recognizes Muscle-Specific kinase (MuSK). 
     In some instances, the binding moiety A is an anti-myosin antibody. In some cases, the anti-myosin antibody is a humanized antibody. In other cases, the anti-myosin antibody is a chimeric antibody. In additional cases, the anti-myosin antibody is a monovalent, a divalent, or a multi-valent antibody. 
     In some instances, the binding moiety A is an anti-transferrin (anti-CD71) antibody. In some cases, the anti-transferrin antibody is a humanized antibody. In other cases, the anti-transferrin antibody is a chimeric antibody. In additional cases, the anti-transferrin antibody is a monovalent, a divalent, or a multi-valent antibody. In some embodiments, exemplary anti-transferrin antibodies include MAB5746 from R&amp;D Systems, AHP858 from Bio-Rad Laboratories, A80-128A from Bethyl Laboratories, Inc., and T2027 from MilliporeSigma. 
     In some instances, the binding moiety A is an antibody that recognizes MuSK. In some cases, the anti-MuSK antibody is a humanized antibody. In other cases, the anti-MuSK antibody is a chimeric antibody. In additional cases, the anti-MuSK antibody is a monovalent, a divalent, or a multi-valent antibody. 
     In some embodiments, the binding moiety A is conjugated to a polynucleic acid molecule (B) non-specifically. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) via a lysine residue or a cysteine residue, in a non-site specific manner. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) via a lysine residue in a non-site specific manner. In some cases, the binding moiety A is conjugated to a polynucleic acid molecule (B) via a cysteine residue in a non-site specific manner. 
     In some embodiments, the binding moiety A is conjugated to a polynucleic acid molecule (B) in a site-specific manner. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) through a lysine residue, a cysteine residue, at the 5′-terminus, at the 3′-terminus, an unnatural amino acid, or an enzyme-modified or enzyme-catalyzed residue, via a site-specific manner. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) through a lysine residue via a site-specific manner. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) through a cysteine residue via a site-specific manner. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) at the 5′-terminus via a site-specific manner. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) at the 3′-terminus via a site-specific manner. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) through an unnatural amino acid via a site-specific manner. In some instances, the binding moiety A is conjugated to a polynucleic acid molecule (B) through an enzyme-modified or enzyme-catalyzed residue via a site-specific manner. 
     In some embodiments, one or more polynucleic acid molecule (B) is conjugated to a binding moiety A. In some instances, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or more polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 1 polynucleic acid molecule is conjugated to one binding moiety A. In some instances, about 2 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 3 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 4 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 5 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 6 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 7 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 8 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 9 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 10 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 11 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 12 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 13 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 14 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 15 polynucleic acid molecules are conjugated to one binding moiety A. In some instances, about 16 polynucleic acid molecules are conjugated to one binding moiety A. In some cases, the one or more polynucleic acid molecules are the same. In other cases, the one or more polynucleic acid molecules are different. 
     In some embodiments, the number of polynucleic acid molecule (B) conjugated to a binding moiety A forms a ratio. In some instances, the ratio is referred to as a DAR (drug-to-antibody) ratio, in which the drug as referred to herein is the polynucleic acid molecule (B). In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 1 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 2 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 3 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 4 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 5 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 6 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 7 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 8 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 9 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 10 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 11 or greater. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 12 or greater. 
     In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 1. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 2. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 3. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 4. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 5. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 6. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 7. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 8. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 9. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 10. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 11. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 12. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 13. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 14. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 15. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is about 16. 
     In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is 1. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is 2. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is 4. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is 6. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is 8. In some instances, the DAR ratio of the polynucleic acid molecule (B) to binding moiety A is 12. 
     In some instances, a conjugate comprising polynucleic acid molecule (B) and binding moiety A has improved activity as compared to a conjugate comprising polynucleic acid molecule (B) without a binding moiety A. In some instances, improved activity results in enhanced biologically relevant functions, e.g., improved stability, affinity, binding, functional activity, and efficacy in treatment or prevention of a disease state. In some instances, the disease state is a result of one or more mutated exons of a gene. In some instances, the conjugate comprising polynucleic acid molecule (B) and binding moiety A results in increased exon skipping of the one or more mutated exons as compared to the conjugate comprising polynucleic acid molecule (B) without a binding moiety A. In some instances, exon skipping is increased by at least or about 5%, 10%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or more than 95% in the conjugate comprising polynucleic acid molecule (B) and binding moiety A as compared to the conjugate comprising polynucleic acid molecule (B) without a binding moiety A. 
     In some embodiments, an antibody or its binding fragment is further modified using conventional techniques known in the art, for example, by using amino acid deletion, insertion, substitution, addition, and/or by recombination and/or any other modification (e.g. posttranslational and chemical modifications, such as glycosylation and phosphorylation) known in the art either alone or in combination. In some instances, the modification further comprises a modification for modulating interaction with Fc receptors. In some instances, the one or more modifications include those described in, for example, International Publication No. WO97/34631, which discloses amino acid residues involved in the interaction between the Fc domain and the FcRn receptor. Methods for introducing such modifications in the nucleic acid sequence underlying the amino acid sequence of an antibody or its binding fragment is well known to the person skilled in the art. 
     In some instances, an antibody binding fragment further encompasses its derivatives and includes polypeptide sequences containing at least one CDR. 
     In some instances, the term “single-chain” as used herein means that the first and second domains of a bi-specific single chain construct are covalently linked, preferably in the form of a co-linear amino acid sequence encodable by a single nucleic acid molecule. 
     In some instances, a bispecific single chain antibody construct relates to a construct comprising two antibody derived binding domains. In such embodiments, bi-specific single chain antibody construct is tandem bi-scFv or diabody. In some instances, a scFv contains a VH and VL domain connected by a linker peptide. In some instances, linkers are of a length and sequence sufficient to ensure that each of the first and second domains can, independently from one another, retain their differential binding specificities. 
     In some embodiments, binding to or interacting with as used herein defines a binding/interaction of at least two antigen-interaction-sites with each other. In some instances, antigen-interaction-site defines a motif of a polypeptide that shows the capacity of specific interaction with a specific antigen or a specific group of antigens. In some cases, the binding/interaction is also understood to define a specific recognition. In such cases, specific recognition refers to that the antibody or its binding fragment is capable of specifically interacting with and/or binding to at least two amino acids of each of a target molecule. For example, specific recognition relates to the specificity of the antibody molecule, or to its ability to discriminate between the specific regions of a target molecule. In additional instances, the specific interaction of the antigen-interaction-site with its specific antigen results in an initiation of a signal, e.g. due to the induction of a change of the conformation of the antigen, an oligomerization of the antigen, etc. In further embodiments, the binding is exemplified by the specificity of a “key-lock-principle”. Thus in some instances, specific motifs in the amino acid sequence of the antigen-interaction-site and the antigen bind to each other as a result of their primary, secondary or tertiary structure as well as the result of secondary modifications of said structure. In such cases, the specific interaction of the antigen-interaction-site with its specific antigen results as well in a simple binding of the site to the antigen. 
     In some instances, specific interaction further refers to a reduced cross-reactivity of the antibody or its binding fragment or a reduced off-target effect. For example, the antibody or its binding fragment that bind to the polypeptide/protein of interest but do not or do not essentially bind to any of the other polypeptides are considered as specific for the polypeptide/protein of interest. Examples for the specific interaction of an antigen-interaction-site with a specific antigen comprise the specificity of a ligand for its receptor, for example, the interaction of an antigenic determinant (epitope) with the antigenic binding site of an antibody. 
     Additional Binding Moieties 
     In some embodiments, the binding moiety is a plasma protein. In some instances, the plasma protein comprises albumin. In some instances, the binding moiety A is albumin. In some instances, albumin is conjugated by one or more of a conjugation chemistry described herein to a polynucleic acid molecule. In some instances, albumin is conjugated by native ligation chemistry to a polynucleic acid molecule. In some instances, albumin is conjugated by lysine conjugation to a polynucleic acid molecule. 
     In some instances, the binding moiety is a steroid. Exemplary steroids include cholesterol, phospholipids, di- and triacylglycerols, fatty acids, hydrocarbons that are saturated, unsaturated, comprise substitutions, or combinations thereof. In some instances, the steroid is cholesterol. In some instances, the binding moiety is cholesterol. In some instances, cholesterol is conjugated by one or more of a conjugation chemistry described herein to a polynucleic acid molecule. In some instances, cholesterol is conjugated by native ligation chemistry to a polynucleic acid molecule. In some instances, cholesterol is conjugated by lysine conjugation to a polynucleic acid molecule. 
     In some instances, the binding moiety is a polymer, including but not limited to polynucleic acid molecule aptamers that bind to specific surface markers on cells. In this instance the binding moiety is a polynucleic acid that does not hybridize to a target gene or mRNA, but instead is capable of selectively binding to a cell surface marker similarly to an antibody binding to its specific epitope of a cell surface marker. 
     In some cases, the binding moiety is a peptide. In some cases, the peptide comprises between about 1 and about 3 kDa. In some cases, the peptide comprises between about 1.2 and about 2.8 kDa, about 1.5 and about 2.5 kDa, or about 1.5 and about 2 kDa. In some instances, the peptide is a bicyclic peptide. In some cases, the bicyclic peptide is a constrained bicyclic peptide. In some instances, the binding moiety is a bicyclic peptide (e.g., bicycles from Bicycle Therapeutics). 
     In additional cases, the binding moiety is a small molecule. In some instances, the small molecule is an antibody-recruiting small molecule. In some cases, the antibody-recruiting small molecule comprises a target-binding terminus and an antibody-binding terminus, in which the target-binding terminus is capable of recognizing and interacting with a cell surface receptor. For example, in some instances, the target-binding terminus comprising a glutamate urea compound enables interaction with PSMA, thereby, enhances an antibody interaction with a cell that expresses PSMA. In some instances, a binding moiety is a small molecule described in Zhang et al., “A remote arene-binding site on prostate specific membrane antigen revealed by antibody-recruiting small molecules,” J Am Chem Soc. 132(36): 12711-12716 (2010); or McEnaney, et al., “Antibody-recruiting molecules: an emerging paradigm for engaging immune function in treating human disease,” ACS Chem Biol. 7(7): 1139-1151 (2012). 
     Conjugation Chemistry 
     In some embodiments, a polynucleic acid molecule B is conjugated to a binding moiety. In some instances, the binding moiety comprises amino acids, peptides, polypeptides, proteins, antibodies, antigens, toxins, hormones, lipids, nucleotides, nucleosides, sugars, carbohydrates, polymers such as polyethylene glycol and polypropylene glycol, as well as analogs or derivatives of all of these classes of substances. Additional examples of binding moiety also include steroids, such as cholesterol, phospholipids, di- and triacylglycerols, fatty acids, hydrocarbons (e.g., saturated, unsaturated, or contains substitutions), enzyme substrates, biotin, digoxigenin, and polysaccharides. In some instances, the binding moiety is an antibody or binding fragment thereof. In some instances, the polynucleic acid molecule is further conjugated to a polymer, and optionally an endosomolytic moiety. 
     In some embodiments, the polynucleic acid molecule is conjugated to the binding moiety by a chemical ligation process. In some instances, the polynucleic acid molecule is conjugated to the binding moiety by a native ligation. In some instances, the conjugation is as described in: Dawson, et al. “Synthesis of proteins by native chemical ligation,”  Science  1994, 266, 776-779; Dawson, et al. “Modulation of Reactivity in Native Chemical Ligation through the Use of Thiol Additives,”  J Am. Chem. Soc.  1997, 119, 4325-4329; Hackeng, et al. “Protein synthesis by native chemical ligation: Expanded scope by using straightforward methodology.,”  Proc. Natl. Acad. Sci. USA  1999, 96, 10068-10073; or Wu, et al. “Building complex glycopeptides: Development of a cysteine-free native chemical ligation protocol,”  Angew. Chem. Int. Ed.  2006, 45, 4116-4125. In some instances, the conjugation is as described in U.S. Pat. No. 8,936,910. In some embodiments, the polynucleic acid molecule is conjugated to the binding moiety either site-specifically or non-specifically via native ligation chemistry. 
     In some instances, the polynucleic acid molecule is conjugated to the binding moiety by a site-directed method utilizing a “traceless” coupling technology (Philochem). In some instances, the “traceless” coupling technology utilizes an N-terminal 1,2-aminothiol group on the binding moiety which is then conjugate with a polynucleic acid molecule containing an aldehyde group. (see Casi et al., “Site-specific traceless coupling of potent cytotoxic drugs to recombinant antibodies for pharmacodelivery,”  JACS  134(13): 5887-5892 (2012)) 
     In some instances, the polynucleic acid molecule is conjugated to the binding moiety by a site-directed method utilizing an unnatural amino acid incorporated into the binding moiety. In some instances, the unnatural amino acid comprises p-acetylphenylalanine (pAcPhe). In some instances, the keto group of pAcPhe is selectively coupled to an alkoxy-amine derivatived conjugating moiety to form an oxime bond. (see Axup et al., “Synthesis of site-specific antibody-drug conjugates using unnatural amino acids,”  PNAS  109(40): 16101-16106 (2012)). 
     In some instances, the polynucleic acid molecule is conjugated to the binding moiety by a site-directed method utilizing an enzyme-catalyzed process. In some instances, the site-directed method utilizes SMARTag™ technology (Redwood). In some instances, the SMARTag™ technology comprises generation of a formylglycine (FGly) residue from cysteine by formylglycine-generating enzyme (FGE) through an oxidation process under the presence of an aldehyde tag and the subsequent conjugation of FGly to an alkylhydraine-functionalized polynucleic acid molecule via hydrazino-Pictet-Spengler (HIPS) ligation. (see Wu et al., “Site-specific chemical modification of recombinant proteins produced in mammalian cells by using the genetically encoded aldehyde tag,”  PNAS  106(9): 3000-3005 (2009); Agarwal, et al., “A Pictet-Spengler ligation for protein chemical modification,”  PNAS  110(1): 46-51 (2013)) 
     In some instances, the enzyme-catalyzed process comprises microbial transglutaminase (mTG). In some cases, the polynucleic acid molecule is conjugated to the binding moiety utilizing a microbial transglutaminze catalyzed process. In some instances, mTG catalyzes the formation of a covalent bond between the amide side chain of a glutamine within the recognition sequence and a primary amine of a functionalized polynucleic acid molecule. In some instances, mTG is produced from  Streptomyces  mobarensis. (see Strop et al., “Location matters: site of conjugation modulates stability and pharmacokinetics of antibody drug conjugates,”  Chemistry and Biology  20(2) 161-167 (2013)) 
     In some instances, the polynucleic acid molecule is conjugated to the binding moiety by a method as described in PCT Publication No. WO2014/140317, which utilizes a sequence-specific transpeptidase. 
     In some instances, the polynucleic acid molecule is conjugated to the binding moiety by a method as described in U.S. Patent Publication Nos. 2015/0105539 and 2015/0105540. 
     Production of Antibodies or Binding Fragments Thereof 
     In some embodiments, polypeptides described herein (e.g., antibodies and its binding fragments) are produced using any method known in the art to be useful for the synthesis of polypeptides (e.g., antibodies), in particular, by chemical synthesis or by recombinant expression, and are preferably produced by recombinant expression techniques. 
     In some instances, an antibody or its binding fragment thereof is expressed recombinantly, and the nucleic acid encoding the antibody or its binding fragment is assembled from chemically synthesized oligonucleotides (e.g., as described in Kutmeier et al., 1994 , BioTechniques  17:242), which involves the synthesis of overlapping oligonucleotides containing portions of the sequence encoding the antibody, annealing and ligation of those oligonucleotides, and then amplification of the ligated oligonucleotides by PCR. 
     Alternatively, a nucleic acid molecule encoding an antibody is optionally generated from a suitable source (e.g., an antibody cDNA library, or cDNA library generated from any tissue or cells expressing the immunoglobulin) by PCR amplification using synthetic primers hybridizable to the 3′ and 5′ ends of the sequence or by cloning using an oligonucleotide probe specific for the particular gene sequence. 
     In some instances, an antibody or its binding is optionally generated by immunizing an animal, such as a rabbit, to generate polyclonal antibodies or, more preferably, by generating monoclonal antibodies, e.g., as described by Kohler and Milstein (1975 , Nature  256:495-497) or, as described by Kozbor et al. (1983 , Immunology Today  4:72) or Cole et al. (1985 in  Monoclonal Antibodies and Cancer Therapy , Alan R. Liss, Inc., pp. 77-96). Alternatively, a clone encoding at least the Fab portion of the antibody is optionally obtained by screening Fab expression libraries (e.g., as described in Huse et al., 1989 , Science  246:1275-1281) for clones of Fab fragments that bind the specific antigen or by screening antibody libraries (See, e.g., Clackson et al., 1991 , Nature  352:624; Hane et al., 1997  Proc. Natl. Acad. Sci. USA  94:4937). 
     In some embodiments, techniques developed for the production of “chimeric antibodies” (Morrison et al., 1984 , Proc. Natl. Acad. Sci.  81:851-855; Neuberger et al., 1984 , Nature  312:604-608; Takeda et al., 1985 , Nature  314:452-454) by splicing genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity are used. A chimeric antibody is a molecule in which different portions are derived from different animal species, such as those having a variable region derived from a murine monoclonal antibody and a human immunoglobulin constant region, e.g., humanized antibodies. 
     In some embodiments, techniques described for the production of single chain antibodies (U.S. Pat. No. 4,694,778; Bird, 1988 , Science  242:423-42; Huston et al., 1988 , Proc. Natl. Acad. Sci. USA  85:5879-5883; and Ward et al., 1989 , Nature  334:544-54) are adapted to produce single chain antibodies. Single chain antibodies are formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge, resulting in a single chain polypeptide. Techniques for the assembly of functional Fv fragments in  E. coli  are also optionally used (Skerra et al., 1988 , Science  242:1038-1041). 
     In some embodiments, an expression vector comprising the nucleotide sequence of an antibody or the nucleotide sequence of an antibody is transferred to a host cell by conventional techniques (e.g., electroporation, liposomal transfection, and calcium phosphate precipitation), and the transfected cells are then cultured by conventional techniques to produce the antibody. In specific embodiments, the expression of the antibody is regulated by a constitutive, an inducible or a tissue, specific promoter. 
     In some embodiments, a variety of host-expression vector systems is utilized to express an antibody or its binding fragment described herein. Such host-expression systems represent vehicles by which the coding sequences of the antibody is produced and subsequently purified, but also represent cells that are, when transformed or transfected with the appropriate nucleotide coding sequences, express an antibody or its binding fragment in situ. These include, but are not limited to, microorganisms such as bacteria (e.g.,  E. coli  and  B. subtilis ) transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing an antibody or its binding fragment coding sequences; yeast (e.g.,  Saccharomyces Pichia ) transformed with recombinant yeast expression vectors containing an antibody or its binding fragment coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing an antibody or its binding fragment coding sequences; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus (CaMV) and tobacco mosaic virus (TMV)) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing an antibody or its binding fragment coding sequences; or mammalian cell systems (e.g., COS, CHO, BH, 293, 293T, 3T3 cells) harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g. the adenovirus late promoter; the vaccinia virus 7.5K promoter). 
     For long-term, high-yield production of recombinant proteins, stable expression is preferred. In some instances, cell lines that stably express an antibody are optionally engineered. Rather than using expression vectors that contain viral origins of replication, host cells are transformed with DNA controlled by appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker. Following the introduction of the foreign DNA, engineered cells are then allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media. The selectable marker in the recombinant plasmid confers resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow to form foci that in turn are cloned and expanded into cell lines. This method can advantageously be used to engineer cell lines which express the antibody or its binding fragments. 
     In some instances, a number of selection systems are used, including but not limited to the herpes simplex virus thymidine kinase (Wigler et al., 1977 , Cell  11:223), hypoxanthine-guanine phosphoribosyltransferase (Szybalska &amp; Szybalski, 192 , Proc. Natl. Acad. Sci. USA  48:202), and adenine phosphoribosyltransferase (Lowy et al., 1980 , Cell  22:817) genes are employed in tk-, hgprt- or aprt-cells, respectively. Also, antimetabolite resistance are used as the basis of selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler et al., 1980 , Proc. Natl. Acad. Sci. USA  77:357; O&#39;Hare et al., 1981 , Proc. Natl. Acad. Sci. USA  78:1527); gpt, which confers resistance to mycophenolic acid (Mulligan &amp; Berg, 1981 , Proc. Natl. Acad. Sci. USA  78:2072); neo, which confers resistance to the aminoglycoside G-418 (Clinical Pharmacy 12:488-505; Wu and Wu, 1991 , Biotherapy  3:87-95; Tolstoshev, 1993 , Ann. Rev. Pharmacol. Toxicol.  32:573-596; Mulligan, 1993 , Science  260:926-932; and Morgan and Anderson, 1993 , Ann. Rev. Biochem.  62:191-217; May, 1993 , TIB TECH  11(5): 155-215) and hygro, which confers resistance to hygromycin (Santerre et al., 1984 , Gene  30:147). Methods commonly known in the art of recombinant DNA technology which can be used are described in Ausubel et al. (eds., 1993 , Current Protocols in Molecular Biology , John Wiley &amp; Sons, NY; Kriegler, 1990, Gene Transfer and Expression,  A Laboratory Manual , Stockton Press, NY; and in Chapters 12 and 13, Dracopoli et al. (eds), 1994 , Current Protocols in Human Genetics , John Wiley &amp; Sons, NY.; Colberre-Garapin et al., 1981 , J. Mol. Biol.  150:1). 
     In some instances, the expression levels of an antibody are increased by vector amplification (for a review, see Bebbington and Hentschel, The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, Vol. 3. (Academic Press, New York, 1987)). When a marker in the vector system expressing an antibody is amplifiable, an increase in the level of inhibitor present in culture of host cell will increase the number of copies of the marker gene. Since the amplified region is associated with the nucleotide sequence of the antibody, production of the antibody will also increase (Crouse et al., 1983 , Mol. Cell Biol.  3:257). 
     In some instances, any method known in the art for purification or analysis of an antibody or antibody conjugates is used, for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after Protein A, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for the purification of proteins. Exemplary chromatography methods included, but are not limited to, strong anion exchange chromatography, hydrophobic interaction chromatography, size exclusion chromatography, and fast protein liquid chromatography. 
     Polymer Conjugating Moiety 
     In some embodiments, a polymer moiety C is further conjugated to a polynucleic acid molecule described herein, a binding moiety described herein, or in combinations thereof. In some instances, a polymer moiety C is conjugated a polynucleic acid molecule. In some cases, a polymer moiety C is conjugated to a binding moiety. In other cases, a polymer moiety C is conjugated to a polynucleic acid molecule-binding moiety molecule. In additional cases, a polymer moiety C is conjugated, as illustrated supra. 
     In some instances, the polymer moiety C is a natural or synthetic polymer, consisting of long chains of branched or unbranched monomers, and/or cross-linked network of monomers in two or three dimensions. In some instances, the polymer moiety C includes a polysaccharide, lignin, rubber, or polyalkylen oxide (e.g., polyethylene glycol). In some instances, the at least one polymer moiety C includes, but is not limited to, alpha-, omega-dihydroxylpolyethyleneglycol, biodegradable lactone-based polymer, e.g. polyacrylic acid, polylactide acid (PLA), poly(glycolic acid) (PGA), polypropylene, polystyrene, polyolefin, polyamide, polycyanoacrylate, polyimide, polyethylenterephthalat (PET, PETG), polyethylene terephthalate (PETE), polytetramethylene glycol (PTG), or polyurethane as well as mixtures thereof. As used herein, a mixture refers to the use of different polymers within the same compound as well as in reference to block copolymers. In some cases, block copolymers are polymers wherein at least one section of a polymer is build up from monomers of another polymer. In some instances, the polymer moiety C comprises polyalkylene oxide. In some instances, the polymer moiety C comprises PEG. In some instances, the polymer moiety C comprises polyethylene imide (PEI) or hydroxy ethyl starch (HES). 
     In some instances, C is a PEG moiety. In some instances, the PEG moiety is conjugated at the 5′ terminus of the polynucleic acid molecule while the binding moiety is conjugated at the 3′ terminus of the polynucleic acid molecule. In some instances, the PEG moiety is conjugated at the 3′ terminus of the polynucleic acid molecule while the binding moiety is conjugated at the 5′ terminus of the polynucleic acid molecule. In some instances, the PEG moiety is conjugated to an internal site of the polynucleic acid molecule. In some instances, the PEG moiety, the binding moiety, or a combination thereof, are conjugated to an internal site of the polynucleic acid molecule. In some instances, the conjugation is a direct conjugation. In some instances, the conjugation is via native ligation. 
     In some embodiments, the polyalkylene oxide (e.g., PEG) is a polydispers or monodispers compound. In some instances, polydispers material comprises disperse distribution of different molecular weight of the material, characterized by mean weight (weight average) size and dispersity. In some instances, the monodisperse PEG comprises one size of molecules. In some embodiments, C is poly- or monodispersed polyalkylene oxide (e.g., PEG) and the indicated molecular weight represents an average of the molecular weight of the polyalkylene oxide, e.g., PEG, molecules. 
     In some embodiments, the molecular weight of the polyalkylene oxide (e.g., PEG) is about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1450, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3250, 3350, 3500, 3750, 4000, 4250, 4500, 4600, 4750, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 10,000, 12,000, 20,000, 35,000, 40,000, 50,000, 60,000, or 100,000 Da. 
     In some embodiments, C is polyalkylene oxide (e.g., PEG) and has a molecular weight of about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1450, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3250, 3350, 3500, 3750, 4000, 4250, 4500, 4600, 4750, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 10,000, 12,000, 20,000, 35,000, 40,000, 50,000, 60,000, or 100,000 Da. In some embodiments, C is PEG and has a molecular weight of about 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1450, 1500, 1600, 1700, 1800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2600, 2700, 2800, 2900, 3000, 3250, 3350, 3500, 3750, 4000, 4250, 4500, 4600, 4750, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 10,000, 12,000, 20,000, 35,000, 40,000, 50,000, 60,000, or 100,000 Da. In some instances, the molecular weight of C is about 200 Da. In some instances, the molecular weight of C is about 300 Da. In some instances, the molecular weight of C is about 400 Da. In some instances, the molecular weight of C is about 500 Da. In some instances, the molecular weight of C is about 600 Da. In some instances, the molecular weight of C is about 700 Da. In some instances, the molecular weight of C is about 800 Da. In some instances, the molecular weight of C is about 900 Da. In some instances, the molecular weight of C is about 1000 Da. In some instances, the molecular weight of C is about 1100 Da. In some instances, the molecular weight of C is about 1200 Da. In some instances, the molecular weight of C is about 1300 Da. In some instances, the molecular weight of C is about 1400 Da. In some instances, the molecular weight of C is about 1450 Da. In some instances, the molecular weight of C is about 1500 Da. In some instances, the molecular weight of C is about 1600 Da. In some instances, the molecular weight of C is about 1700 Da. In some instances, the molecular weight of C is about 1800 Da. In some instances, the molecular weight of C is about 1900 Da. In some instances, the molecular weight of C is about 2000 Da. In some instances, the molecular weight of C is about 2100 Da. In some instances, the molecular weight of C is about 2200 Da. In some instances, the molecular weight of C is about 2300 Da. In some instances, the molecular weight of C is about 2400 Da. In some instances, the molecular weight of C is about 2500 Da. In some instances, the molecular weight of C is about 2600 Da. In some instances, the molecular weight of C is about 2700 Da. In some instances, the molecular weight of C is about 2800 Da. In some instances, the molecular weight of C is about 2900 Da. In some instances, the molecular weight of C is about 3000 Da. In some instances, the molecular weight of C is about 3250 Da. In some instances, the molecular weight of C is about 3350 Da. In some instances, the molecular weight of C is about 3500 Da. In some instances, the molecular weight of C is about 3750 Da. In some instances, the molecular weight of C is about 4000 Da. In some instances, the molecular weight of C is about 4250 Da. In some instances, the molecular weight of C is about 4500 Da. In some instances, the molecular weight of C is about 4600 Da. In some instances, the molecular weight of C is about 4750 Da. In some instances, the molecular weight of C is about 5000 Da. In some instances, the molecular weight of C is about 5500 Da. In some instances, the molecular weight of C is about 6000 Da. In some instances, the molecular weight of C is about 6500 Da. In some instances, the molecular weight of C is about 7000 Da. In some instances, the molecular weight of C is about 7500 Da. In some instances, the molecular weight of C is about 8000 Da. In some instances, the molecular weight of C is about 10,000 Da. In some instances, the molecular weight of C is about 12,000 Da. In some instances, the molecular weight of C is about 20,000 Da. In some instances, the molecular weight of C is about 35,000 Da. In some instances, the molecular weight of C is about 40,000 Da. In some instances, the molecular weight of C is about 50,000 Da. In some instances, the molecular weight of C is about 60,000 Da. In some instances, the molecular weight of C is about 100,000 Da. 
     In some embodiments, the polyalkylene oxide (e.g., PEG) is a discrete PEG, in which the discrete PEG is a polymeric PEG comprising more than one repeating ethylene oxide units. In some instances, a discrete PEG (dPEG) comprises from 2 to 60, from 2 to 50, or from 2 to 48 repeating ethylene oxide units. In some instances, a dPEG comprises about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 22, 24, 26, 28, 30, 35, 40, 42, 48, 50 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 2 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 3 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 4 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 5 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 6 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 7 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 8 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 9 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 10 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 11 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 12 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 13 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 14 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 15 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 16 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 17 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 18 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 19 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 20 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 22 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 24 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 26 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 28 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 30 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 35 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 40 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 42 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 48 or more repeating ethylene oxide units. In some instances, a dPEG comprises about 50 or more repeating ethylene oxide units. In some cases, a dPEG is synthesized as a single molecular weight compound from pure (e.g., about 95%, 98%, 99%, or 99.5%) staring material in a step-wise fashion. In some cases, a dPEG has a specific molecular weight, rather than an average molecular weight. In some cases, a dPEG described herein is a dPEG from Quanta Biodesign, LMD. 
     In some embodiments, the polymer moiety C comprises a cationic mucic acid-based polymer (cMAP). In some instances, cMAP comprises one or more subunit of at least one repeating subunit, and the subunit structure is represented as Formula (V): 
     
       
         
         
             
             
         
       
     
     wherein m is independently at each occurrence 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, preferably 4-6 or 5; and n is independently at each occurrence 1, 2, 3, 4, or 5. In some embodiments, m and n are, for example, about 10. 
     In some instances, cMAP is further conjugated to a PEG moiety, generating a cMAP-PEG copolymer, an mPEG-cMAP-PEGm triblock polymer, or a cMAP-PEG-cMAP triblock polymer. In some instances, the PEG moiety is in a range of from about 500 Da to about 50,000 Da. In some instances, the PEG moiety is in a range of from about 500 Da to about 1000 Da, greater than 1000 Da to about 5000 Da, greater than 5000 Da to about 10,000 Da, greater than 10,000 to about 25,000 Da, greater than 25,000 Da to about 50,000 Da, or any combination of two or more of these ranges. 
     In some instances, the polymer moiety C is cMAP-PEG copolymer, an mPEG-cMAP-PEGm triblock polymer, or a cMAP-PEG-cMAP triblock polymer. In some cases, the polymer moiety C is cMAP-PEG copolymer. In other cases, the polymer moiety C is an mPEG-cMAP-PEGm triblock polymer. In additional cases, the polymer moiety C is a cMAP-PEG-cMAP triblock polymer. 
     In some embodiments, the polymer moiety C is conjugated to the polynucleic acid molecule, the binding moiety, and optionally to the endosomolytic moiety as illustrated supra. 
     Endosomolytic Moiety 
     In some embodiments, a molecule of Formula (I): A-X—B—Y—C, further comprises an additional conjugating moiety. In some instances, the additional conjugating moiety is an endosomolytic moiety. In some cases, the endosomolytic moiety is a cellular compartmental release component, such as a compound capable of releasing from any of the cellular compartments known in the art, such as the endosome, lysosome, endoplasmic reticulum (ER), golgi apparatus, microtubule, peroxisome, or other vesicular bodies with the cell. In some cases, the endosomolytic moiety comprises an endosomolytic polypeptide, an endosomolytic polymer, an endosomolytic lipid, or an endosomolytic small molecule. In some cases, the endosomolytic moiety comprises an endosomolytic polypeptide. In other cases, the endosomolytic moiety comprises an endosomolytic polymer. 
     Endosomolytic Polypeptides 
     In some embodiments, a molecule of Formula (I): A-X—B—Y—C, is further conjugated with an endosomolytic polypeptide. In some embodiments, a molecule of Formula (V): A-(X 1 —B) n  or Formula (II): A-X 1 —(B—X 2 —C) n  is further conjugated with an endosomolytic polypeptide. In some cases, the endosomolytic polypeptide is a pH-dependent membrane active peptide. In some cases, the endosomolytic polypeptide is an amphipathic polypeptide. In additional cases, the endosomolytic polypeptide is a peptidomimetic. In some instances, the endosomolytic polypeptide comprises INF, melittin, meucin, or their respective derivatives thereof. In some instances, the endosomolytic polypeptide comprises INF or its derivatives thereof. In other cases, the endosomolytic polypeptide comprises melittin or its derivatives thereof. In additional cases, the endosomolytic polypeptide comprises meucin or its derivatives thereof. 
     In some instances, INF7 is a 24 residue polypeptide those sequence comprises CGIFGEIEELIEEGLENLIDWGNA (SEQ ID NO: 1), or GLFEAIEGFIENGWEGMIDGWYGC (SEQ ID NO: 2). In some instances, INF7 or its derivatives comprise a sequence of: GLFEAIEGFIENGWEGMIWDYGSGSCG (SEQ ID NO: 3), GLFEAIEGFIENGWEGMIDG WYG-(PEG)6-NH2 (SEQ ID NO: 4), or GLFEAIEGFIENGWEGMIWDYG-SGSC-K(GalNAc)2 (SEQ ID NO: 5). 
     In some cases, melittin is a 26 residue polypeptide those sequence comprises CLIGAILKVLATGLPTLISWIKNKRKQ (SEQ ID NO: 6), or GIGAVLKVLTTGLPAISWIKRKRQQ (SEQ ID NO: 7). In some instances, melittin comprises a polypeptide sequence as described in U.S. Pat. No. 8,501,930. 
     In some instances, meucin is an antimicrobial peptide (AMP) derived from the venom gland of the scorpion Mesobuthus eupeus. In some instances, meucin comprises of meucin-13 those sequence comprises IFGAIAGLLKNIF-NH 2  (SEQ ID NO: 8) and meucin-18 those sequence comprises FFGHLFKLATKIIPSLFQ (SEQ ID NO: 9). 
     In some instances, the endosomolytic polypeptide comprises a polypeptide in which its sequence is at least 50%, 60%, 70%, 80%, 90%, 95%, or 99% sequence identity to INF7 or its derivatives thereof, melittin or its derivatives thereof, or meucin or its derivatives thereof. In some instances, the endosomolytic moiety comprises INF7 or its derivatives thereof, melittin or its derivatives thereof, or meucin or its derivatives thereof. 
     In some instances, the endosomolytic moiety is INF7 or its derivatives thereof. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 1-5. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 1. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 2-5. In some cases, the endosomolytic moiety comprises SEQ ID NO: 1. In some cases, the endosomolytic moiety comprises SEQ ID NO: 2-5. In some cases, the endosomolytic moiety consists of SEQ ID NO: 1. In some cases, the endosomolytic moiety consists of SEQ ID NO: 2-5. 
     In some instances, the endosomolytic moiety is melittin or its derivatives thereof. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 6 or 7. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 6. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 7. In some cases, the endosomolytic moiety comprises SEQ ID NO: 6. In some cases, the endosomolytic moiety comprises SEQ ID NO: 7. In some cases, the endosomolytic moiety consists of SEQ ID NO: 6. In some cases, the endosomolytic moiety consists of SEQ ID NO: 7. 
     In some instances, the endosomolytic moiety is meucin or its derivatives thereof. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NOs: 8 or 9. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 8. In some cases, the endosomolytic moiety comprises a polypeptide having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to SEQ ID NO: 9. In some cases, the endosomolytic moiety comprises SEQ ID NO: 8. In some cases, the endosomolytic moiety comprises SEQ ID NO: 9. In some cases, the endosomolytic moiety consists of SEQ ID NO: 8. In some cases, the endosomolytic moiety consists of SEQ ID NO: 9. 
     In some instances, the endosomolytic moiety comprises a sequence as illustrated in Table 1 below. 
     
       
         
           
               
               
               
               
               
             
               
                   
               
               
                   
                   
                   
                 SEQ 
                   
               
               
                   
                   
                   
                 ID 
                   
               
               
                 Name 
                 Origin 
                 Amino Acid Sequence 
                 NO: 
                 Type 
               
               
                   
               
             
            
               
                 Pep-1 
                 NLS from Simian Virus 
                 KETWWETWWTEWSQPKKKR 
                 10 
                 Primary 
               
               
                   
                 40 large antigen and 
                 KV 
                   
                 amphipathic 
               
               
                   
                 Reverse transcriptase 
                   
                   
                   
               
               
                   
                 of HIV 
                   
                   
                   
               
               
                   
               
               
                 pVEC 
                 VE-cadherin 
                 LLIILRRRRIRKQAHAHSK 
                 11 
                 Primary 
               
               
                   
                   
                   
                   
                 amphipathic 
               
               
                   
               
               
                 VT5 
                 Synthetic peptide 
                 DPKGDPKGVTVTVTVTVTGK 
                 12 
                 β-sheet 
               
               
                   
                   
                 GDPKPD 
                   
                 amphipathic 
               
               
                   
               
               
                 C105Y 
                 1-antitrypsin 
                 CSIPPEVKFNKPFVYLI 
                 13 
                 — 
               
               
                   
               
               
                 Transportan 
                 Galanin and mastoparan 
                 GWTLNSAGYLLGKINLKALA 
                 14 
                 Primary 
               
               
                   
                   
                 ALAKKIL 
                   
                 amphipathic 
               
               
                   
               
               
                 TP10 
                 Galanin and mastoparan 
                 AGYLLGKINLKALAALAKKIL 
                 15 
                 Primary 
               
               
                   
                   
                   
                   
                 amphipathic 
               
               
                   
               
               
                 MPG 
                 A hydrofobic domain 
                 GALFLGFLGAAGSTMGA 
                 16 
                 β-sheet 
               
               
                   
                 from the fusion 
                   
                   
                 amphipathic 
               
               
                   
                 sequence of HIV gp41 
                   
                   
                   
               
               
                   
                 and NLS of SV40 T 
                   
                   
                   
               
               
                   
                 antigen 
                   
                   
                   
               
               
                   
               
               
                 gH625 
                 Glycoprotein gH of 
                 HGLASTLTRWAHYNALIRAF 
                 17 
                 Secondary 
               
               
                   
                 HSV type I 
                   
                   
                 amphipathic α- 
               
               
                   
                   
                   
                   
                 helical 
               
               
                   
               
               
                 CADY 
                 PPTG1 peptide 
                 GLWRALWRLLRSLWRLLWRA 
                 18 
                 Secondary 
               
               
                   
                   
                   
                   
                 amphipathic α- 
               
               
                   
                   
                   
                   
                 helical 
               
               
                   
               
               
                 GALA 
                 Synthetic peptide 
                 WEAALAEALAEALAEHLAEA 
                 19 
                 Secondary 
               
               
                   
                   
                 LAEALEALAA 
                   
                 amphipathic α- 
               
               
                   
                   
                   
                   
                 helical 
               
               
                   
               
               
                 INF 
                 Influenza HA2 fusion 
                 GLFEAIEGFIENGWEGMIDGW 
                 20 
                 Secondary 
               
               
                   
                 peptide 
                 YGC 
                   
                 amphipathic α- 
               
               
                   
                   
                   
                   
                 helical/pH- 
               
               
                   
                   
                   
                   
                 dependent 
               
               
                   
                   
                   
                   
                 membrane 
               
               
                   
                   
                   
                   
                 active peptide 
               
               
                   
               
               
                 HA2E5- 
                 Influenza HA2 subunit 
                 GLFGAIAGFIENGWEGMIDGW 
                 21 
                 Secondary 
               
               
                 TAT 
                 of influenza virus X31 
                 YG 
                   
                 amphipathic α- 
               
               
                   
                 strain fusion peptide 
                   
                   
                 helical/pH- 
               
               
                   
                   
                   
                   
                 dependent 
               
               
                   
                   
                   
                   
                 membrane 
               
               
                   
                   
                   
                   
                 active peptide 
               
               
                   
               
               
                 HA2- 
                 Influenza HA2 subunit 
                 GLFGAIAGFIENGWEGMIDGR 
                 22 
                 pH-dependent 
               
               
                 penetratin 
                 of influenza virus X31 
                 QIKIWFQNRRMKW 
                   
                 membrane 
               
               
                   
                 strain fusion peptide 
                 KK-amide 
                   
                 active peptide 
               
               
                   
               
               
                 HA-K4 
                 Influenza HA2 subunit 
                 GLFGAIAGFIENGWEGMIDG- 
                 23 
                 pH-dependent 
               
               
                   
                 of influenza virus X31 
                 SSKKKK 
                   
                 membrane 
               
               
                   
                 strain fusion peptide 
                   
                   
                 active peptide 
               
               
                   
               
               
                 HA2E4 
                 Influenza HA2 subunit 
                 GLFEAIAGFIENGWEGMIDGG 
                 24 
                 pH-dependent 
               
               
                   
                 of influenza virus X31 
                 GYC 
                   
                 membrane 
               
               
                   
                 strain fusion peptide 
                   
                   
                 active peptide 
               
               
                   
               
               
                 H5WYG 
                 HA2 analogue 
                 GLFHAIAHFIHGGWH 
                 25 
                 pH-dependent 
               
               
                   
                   
                 GLIHGWYG 
                   
                 membrane 
               
               
                   
                   
                   
                   
                 active peptide 
               
               
                   
               
               
                 GALA- 
                 INF3 fusion peptide 
                 GLFEAIEGFIENGWEGLAEALA 
                 26 
                 pH-dependent 
               
               
                 INF3- 
                   
                 EALEALAA- 
                   
                 membrane 
               
               
                 (PEG)6-NH 
                   
                 (PEG)6-NH2 
                   
                 active peptide 
               
               
                   
               
               
                 CM18- 
                 Cecropin-A-Melittin 2-12   
                 KWKLFKKIGAVLKVLTTG- 
                 27 
                 pH-dependent 
               
               
                 TAT11 
                 (CM 18 ) fusion peptide 
                 YGRKKRRQRRR 
                   
                 membrane 
               
               
                   
                   
                   
                   
                 active peptide 
               
               
                   
               
            
           
         
       
     
     In some cases, the endosomolytic moiety comprises a Bak BH3 polypeptide which induces apoptosis through antagonization of suppressor targets such as Bcl-2 and/or Bcl-x L . In some instances, the endosomolytic moiety comprises a Bak BH3 polypeptide described in Albarran, et al., “Efficient intracellular delivery of a pro-apoptotic peptide with a pH-responsive carrier,”  Reactive  &amp;  Functional Polymers  71: 261-265 (2011). 
     In some instances, the endosomolytic moiety comprises a polypeptide (e.g., a cell-penetrating polypeptide) as described in PCT Publication Nos. WO2013/166155 or WO2015/069587. 
     Endosomolytic Polymers 
     In some embodiments, a molecule of Formula (V): A-(X 1 —B), or Formula (VI): A-X 1 —(B—X 2 —C) n  is further conjugated with an endosomolytic polymer. As used herein, an endosomolytic polymer comprises a linear, a branched network, a star, a comb, or a ladder type of polymer. In some instances, an endosomolytic polymer is a homopolymer or a copolymer comprising two ro more different types of monomers. In some cases, an endosomolytic polymer is a polycation polymer. In other cases, an endosomolytic polymer is a polyanion polymer. 
     In some instances, a polycation polymer comprises monomer units that are charge positive, charge neutral, or charge negative, with a net charge being positive. In other cases, a polycation polymer comprises a non-polymeric molecule that contains two or more positive charges. Exemplary cationic polymers include, but are not limited to, poly(L-lysine) (PLL), poly(L-arginine) (PLA), polyethyleneimine (PEI), poly[α-(4-aminobutyl)-L-glycolic acid] (PAGA), 2-(dimethylamino)ethyl methacrylate (DMAEMA), or N,N-Diethylaminoethyl Methacrylate (DEAEMA). 
     In some cases, a polyanion polymer comprises monomer units that are charge positive, charge neutral, or charge negative, with a net charge being negative. In other cases, a polyanion polymer comprises a non-polymeric molecule that contains two or more negative charges. Exemplary anionic polymers include p(alkylacrylates) (e.g., poly(propyl acrylic acid) (PPAA)) or poly(N-isopropylacrylamide) (NIPAM). Additional examples include PP75, a L-phenylalanine-poly(L-lysine isophthalamide) polymer described in Khormaee, et al., “Edosomolytic anionic polymer for the cytoplasmic delivery of siRNAs in localized in vivo applications,”  Advanced Functional Materials  23: 565-574 (2013). 
     In some embodiments, an endosomolytic polymer described herein is a pH-responsive endosomolytic polymer. A pH-responsive polymer comprises a polymer that increases in size (swell) or collapses depending on the pH of the environment. Polyacrylic acid and chitosan are examples of pH-responsive polymers. 
     In some instances, an endosomolytic moiety described herein is a membrane-disruptive polymer. In some cases, the membrane-disruptive polymer comprises a cationic polymer, a neutral or hydrophobic polymer, or an anionic polymer. In some instances, the membrane-disruptive polymer is a hydrophilic polymer. 
     In some instances, an endosomolytic moiety described herein is a pH-responsive membrane-disruptive polymer. Exemplary pH-responsive membrane-disruptive polymers include p(alkylacrylic acids), poly(N-isopropylacrylamide) (NIPAM) copolymers, succinylated p(glycidols), and p(P3-malic acid) polymers. 
     In some instances, p(alkylacrylic acids) include poly(propylacrylic acid) (polyPAA), poly(methacrylic acid) (PMAA), poly(ethylacrylic acid) (PEAA), and poly(propyl acrylic acid) (PPAA). In some instances, a p(alkylacrylic acid) include a p(alkylacrylic acid) described in Jones, et al.,  Biochemistry Journal  372: 65-75 (2003). 
     In some embodiments, a pH-responsive membrane-disruptive polymer comprises p(butyl acrylate-co-methacrylic acid). (see Bulmus, et al.,  Journal of Controlled Release  93: 105-120 (2003); and Yessine, et al.,  Biochimica et Biophysica Acta  1613: 28-38 (2003)) 
     In some embodiments, a pH-responsive membrane-disruptive polymer comprises p(styrene-alt-maleic anhydride). (see Henry, et al.,  Biomacromolecules  7: 2407-2414 (2006)) 
     In some embodiments, a pH-responsive membrane-disruptive polymer comprises pyridyldisulfide acrylate (PDSA) polymers such as poly(MAA-co-PDSA), poly(EAA-co-PDSA), poly(PAA-co-PDSA), poly(MAA-co-BA-co-PDSA), poly(EAA-co-BA-co-PDSA), or poly(PAA-co-BA-co-PDSA) polymers. (see El-Sayed, et al., “Rational design of composition and activity correlations for pH-responsive and glutathione-reactive polymer therapeutics,”  Journal of Controlled Release  104: 417-427 (2005); or Flanary et al., “Antigen delivery with poly(propylacrylic acid) conjugation enhanced MHC-1 presentation and T-cell activation,”  Bioconjugate Chem.  20: 241-248 (2009)) 
     In some embodiments, a pH-responsive membrane-disruptive polymer comprises a lytic polymer comprising the base structure of: 
     
       
         
         
             
             
         
       
     
     In some instances, an endosomolytic moiety described herein is further conjugated to an additional conjugate, e.g., a polymer (e.g., PEG), or a modified polymer (e.g., cholesterol-modified polymer). 
     In some instances, the additional conjugate comprises a detergent (e.g., Triton X-100). In some instances, an endosomolytic moiety described herein comprises a polymer (e.g., a poly(amidoamine)) conjugated with a detergent (e.g., Triton X-100). In some instances, an endosomolytic moiety described herein comprises poly(amidoamine)-Triton X-100 conjugate (Duncan, et al., “A polymer-Triton X-100 conjugate capable of pH-dependent red blood cell lysis: a model system illustrating the possibility of drug delivery within acidic intracellular compartments,”  Journal of Drug Targeting  2: 341-347 (1994)). 
     Endosomolytic Lipids 
     In some embodiments, the endosomolytic moiety is a lipid (e.g., a fusogenic lipid). In some embodiments, a molecule of Formula (V): A-(X 1 —B), or Formula (VI): A-X 1 —(B—X 2 —C) n  is further conjugated with an endosomolytic lipid (e.g., fusogenic lipid). Exemplary fusogenic lipids include 1,2-dileoyl-sn-3-phosphoethanolamine (DOPE), phosphatidylethanolamine (POPE), palmitoyloleoylphosphatidylcholine (POPC), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-ol (Di-Lin), N-methyl(2,2-di((9Z,12Z)-octadeca-9,12-dienyl)-1,3-dioxolan-4-yl)methanamine (DLin-k-DMA) and N-methyl-2-(2,2-di((9Z,12Z)-octadeca-9,12-dienyl)-1,3-dioxolan-4-yl)ethanamine (XTC). 
     In some instances, an endosomolytic moiety is a lipid (e.g., a fusogenic lipid) described in PCT Publication No. WO09/126,933. 
     Endosomolytic Small Molecules 
     In some embodiments, the endosomolytic moiety is a small molecule. In some embodiments, a molecule of Formula (I): A-(X 1 —B), or Formula (II): A-X 1 —(B—X 2 —C) n  is further conjugated with an endosomolytic small molecule. Exemplary small molecules suitable as endosomolytic moieties include, but are not limited to, quinine, chloroquine, hydroxychloroquines, amodiaquins (carnoquines), amopyroquines, primaquines, mefloquines, nivaquines, halofantrines, quinone imines, or a combination thereof. In some instances, quinoline endosomolytic moieties include, but are not limited to, 7-chloro-4-(4-diethylamino-1-methylbutyl-amino)quinoline (chloroquine); 7-chloro-4-(4-ethyl-(2-hydroxyethyl)-amino-1-methylbutyl-amino)quinoline (hydroxychloroquine); 7-fluoro-4-(4-diethylamino-1-methylbutyl-amino)quinoline; 4-(4-diethylamino-1-methylbutylamino) quinoline; 7-hydroxy-4-(4-diethyl-amino-1-methylbutylamino)quinoline; 7-chloro-4-(4-diethylamino-1-butylamino)quinoline (desmethylchloroquine); 7-fluoro-4-(4-diethylamino-1-butylamino)quinoline); 4-(4-diethyl-amino-1-butylamino)quinoline; 7-hydroxy-4-(4-diethylamino-1-butylamino)quinoline; 7-chloro-4-(1-carboxy-4-diethylamino-1-butylamino)quinoline; 7-fluoro-4-(1-carboxy-4-diethyl-amino-1-butylamino)quinoline; 4-(1-carboxy-4-diethylamino-1-butylamino) quinoline; 7-hydroxy-4-(1-carboxy-4-diethylamino-1-butylamino)quinoline; 7-chloro-4-(1-carboxy-4-diethylamino-1-methylbutylamino)quinoline; 7-fluoro-4-(1-carboxy-4-diethyl-amino-1-methylbutylamino)quinoline; 4-(1-carboxy-4-diethylamino-1-methylbutylamino)quinoline; 7-hydroxy-4-(1-carboxy-4-diethylamino-1-methylbutylamino)quinoline; 7-fluoro-4-(4-ethyl-(2-hydroxyethyl)-amino-1-methylbutylamino)quinoline; 4-(4-ethyl-(2-hydroxy-ethyl)-amino-1-methylbutylamino-)quinoline; 7-hydroxy-4-(4-ethyl-(2-hydroxyethyl)-amino-1-methylbutylamino)quinoline; hydroxychloroquine phosphate; 7-chloro-4-(4-ethyl-(2-hydroxyethyl-1)-amino-1-butylamino)quinoline (desmethylhydroxychloroquine); 7-fluoro-4-(4-ethyl-(2-hydroxyethyl)-amino-1-butylamino)quinoline; 4-(4-ethyl-(2-hydroxyethyl)-amino-1-butylamino)quinoline; 7-hydroxy-4-(4-ethyl-(2-hydroxyethyl)-amino-1-butylamino) quinoline; 7-chloro-4-(1-carboxy-4-ethyl-(2-hydroxyethyl)-amino-1-butylamino)quinoline; 7-fluoro-4-(1-carboxy-4-ethyl-(2-hydroxyethyl)-amino-1-butylamino)quinoline; 4-(1-carboxy-4-ethyl-(2-hydroxyethyl)-amino-1-butylamino)quinoline; 7-hydroxy-4-(1-carboxy-4-ethyl-(2-hydroxyethyl)-amino-1-butylamino)quinoline; 7-chloro-4-(1-carboxy-4-ethyl-(2-hydroxyethyl)-amino-1-methylbutylamino)quinoline; 7-fluoro-4-(1-carboxy-4-ethyl-(2-hydroxyethyl)-amino-1-methylbutylamino)quinoline; 4-(1-carboxy-4-ethyl-(2-hydroxyethyl)-amino-1-methylbutylamino)quinoline; 7-hydroxy-4-(1-carboxy-4-ethyl-(2-hydroxyethyl)-amino-1-methylbutylamino)quinoline; 8-[(4-aminopentyl)amino-6-methoxydihydrochloride quinoline; 1-acetyl-1,2,3,4-tetrahydroquinoline; 8-[(4-aminopentyl)amino]-6-methoxyquinoline dihydrochloride; 1-butyryl-1,2,3,4-tetrahydroquinoline; 3-chloro-4-(4-hydroxy-alpha,alpha′-bis(2-methyl-1-pyrrolidinyl)-2,5-xylidinoquinoline, 4-[(4-diethyl-amino)-1-methylbutyl-amino]-6-methoxyquinoline; 3-fluoro-4-(4-hydroxy-alpha,alpha′-bis(2-methyl-1-pyrrolidinyl)-2,5-xylidinoquinoline, 4-[(4-diethylamino)-1-methylbutyl-amino]-6-methoxyquinoline; 4-(4-hydroxy-alpha,alpha′-bis(2-methyl-1-pyrrolidinyl)-2,5-xylidinoquinoline; 4-[(4-diethylamino)-1-methylbutyl-amino]-6-methoxyquinoline; 3,4-dihydro-1-(2H)-quinolinecarboxyaldehyde; 11′-pentamethylene diquinoleinium diiodide; 8-quinolinol sulfate and amino, aldehyde, carboxylic, hydroxyl, halogen, keto, sulfhydryl and vinyl derivatives or analogs thereof. In some instances, an endosomolytic moiety is a small molecule described in Naisbitt et al (1997, J Pharmacol Exp Therapy 280:884-893) and in U.S. Pat. No. 5,736,557. 
     In some embodiments, the endosomolytic moiety is nigericin or a conjugate thereof, e.g., such as a folate-nigericin ester conjugate, a folate-nigericin amide conjugate, or a folate-nigericin carbamate conjugate. In some instances, the endosomolytic moiety is nigericin described in Rangasamy, et. al., “New mechanism for release of endosomal contents: osmotic lysis via nigericin-mediated K+/H+ exchange,”  Bioconjugate Chem.  29:1047-1059 (2018). 
     Linkers 
     In some embodiments, a linker described herein is a cleavable linker or a non-cleavable linker. In some instances, the linker is a cleavable linker. In other instances, the linker is a non-cleavable linker. 
     In some cases, the linker is a non-polymeric linker. A non-polymeric linker refers to a linker that does not contain a repeating unit of monomers generated by a polymerization process. Exemplary non-polymeric linkers include, but are not limited to, C 1 -C 6  alkyl group (e.g., a C 5 , C 4 , C 3 , C 2 , or C 1  alkyl group), homobifunctional cross linkers, heterobifunctional cross linkers, peptide linkers, traceless linkers, self-immolative linkers, maleimide-based linkers, or combinations thereof. In some cases, the non-polymeric linker comprises a C 1 -C 6  alkyl group (e.g., a C 5 , C 4 , C 3 , C 2 , or C 1  alkyl group), a homobifunctional cross linker, a heterobifunctional cross linker, a peptide linker, a traceless linker, a self-immolative linker, a maleimide-based linker, or a combination thereof. In additional cases, the non-polymeric linker does not comprise more than two of the same type of linkers, e.g., more than two homobifunctional cross linkers, or more than two peptide linkers. In further cases, the non-polymeric linker optionally comprises one or more reactive functional groups. 
     In some instances, the non-polymeric linker does not encompass a polymer that is described above. In some instances, the non-polymeric linker does not encompass a polymer encompassed by the polymer moiety C. In some cases, the non-polymeric linker does not encompass a polyalkylene oxide (e.g., PEG). In some cases, the non-polymeric linker does not encompass a PEG. 
     In some instances, the linker comprises a homobifunctional linker. Exemplary homobifunctional linkers include, but are not limited to, Lomant&#39;s reagent dithiobis (succinimidylpropionate) DSP, 3′3′-dithiobis(sulfosuccinimidyl proprionate (DTSSP), disuccinimidyl suberate (DSS), bis(sulfosuccinimidyl)suberate (BS), disuccinimidyl tartrate (DST), disulfosuccinimidyl tartrate (sulfo DST), ethylene glycobis(succinimidylsuccinate) (EGS), disuccinimidyl glutarate (DSG), N,N′-disuccinimidyl carbonate (DSC), dimethyl adipimidate (DMA), dimethyl pimelimidate (DMP), dimethyl suberimidate (DMS), dimethyl-3,3′-dithiobispropionimidate (DTBP), 1,4-di-3′-(2′-pyridyldithio)propionamido)butane (DPDPB), bismaleimidohexane (BMH), aryl halide-containing compound (DFDNB), such as e.g. 1,5-difluoro-2,4-dinitrobenzene or 1,3-difluoro-4,6-dinitrobenzene, 4,4′-difluoro-3,3′-dinitrophenylsulfone (DFDNPS), bis-[3-(4-azidosalicylamido)ethyl]disulfide (BASED), formaldehyde, glutaraldehyde, 1,4-butanediol diglycidyl ether, adipic acid dihydrazide, carbohydrazide, o-toluidine, 3,3′-dimethylbenzidine, benzidine, α,α′-p-diaminodiphenyl, diiodo-p-xylene sulfonic acid, N,N′-ethylene-bis(iodoacetamide), or N,N′-hexamethylene-bis(iodoacetamide). 
     In some embodiments, the linker comprises a heterobifunctional linker. Exemplary heterobifunctional linker include, but are not limited to, amine-reactive and sulfhydryl cross-linkers such as N-succinimidyl 3-(2-pyridyldithio)propionate (sPDP), long-chain N-succinimidyl 3-(2-pyridyldithio)propionate (LC-sPDP), water-soluble-long-chain N-succinimidyl 3-(2-pyridyldithio) propionate (sulfo-LC-sPDP), succinimidyloxycarbonyl-a-methyl-a-(2-pyridyldithio)toluene (sMPT), sulfosuccinimidyl-6-[α-methyl-α-(2-pyridyldithio)toluamido]hexanoate (sulfo-LC-sMPT), succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sMCC), sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-sMCC), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBs), m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester (sulfo-MBs), N-succinimidyl(4-iodoacteyl)aminobenzoate (sIAB), sulfosuccinimidyl(4-iodoacteyl)aminobenzoate (sulfo-sIAB), succinimidyl-4-(p-maleimidophenyl)butyrate (sMPB), sulfosuccinimidyl-4-(p-maleimidophenyl)butyrate (sulfo-sMPB), N-(y-maleimidobutyryloxy)succinimide ester (GMBs), N-(y-maleimidobutyryloxy)sulfosuccinimide ester (sulfo-GMBs), succinimidyl 6-((iodoacetyl)amino)hexanoate (sIAX), succinimidyl 6-[6-(((iodoacetyl)amino)hexanoyl)amino]hexanoate (sIAXX), succinimidyl 4-(((iodoacetyl)amino)methyl)cyclohexane-1-carboxylate (sIAC), succinimidyl 6-((((4-iodoacetyl)amino)methyl)cyclohexane-1-carbonyl)amino) hexanoate (sIACX), p-nitrophenyl iodoacetate (NPIA), carbonyl-reactive and sulfhydryl-reactive cross-linkers such as 4-(4-N-maleimidophenyl)butyric acid hydrazide (MPBH), 4-(N-maleimidomethyl)cyclohexane-1-carboxyl-hydrazide-8 (M 2 C2H), 3-(2-pyridyldithio)propionyl hydrazide (PDPH), amine-reactive and photoreactive cross-linkers such as N-hydroxysuccinimidyl-4-azidosalicylic acid (NHs-AsA), N-hydroxysulfosuccinimidyl-4-azidosalicylic acid (sulfo-NHs-AsA), sulfosuccinimidyl-(4-azidosalicylamido)hexanoate (sulfo-NHs-LC-AsA), sulfosuccinimidyl-2-(p-azidosalicylamido)ethyl-1,3′-dithiopropionate (sAsD), N-hydroxysuccinimidyl-4-azidobenzoate (HsAB), N-hydroxysulfosuccinimidyl-4-azidobenzoate (sulfo-HsAB), N-succinimidyl-6-(4′-azido-2′-nitrophenylamino)hexanoate (sANPAH), sulfosuccinimidyl-6-(4′-azido-2′-nitrophenylamino)hexanoate (sulfo-sANPAH), N-5-azido-2-nitrobenzoyloxysuccinimide (ANB-NOs), sulfosuccinimidyl-2-(m-azido-o-nitrobenzamido)-ethyl-1,3′-dithiopropionate (sAND), N-succinimidyl-4(4-azidophenyl) 1,3′-dithiopropionate (sADP), N-sulfosuccinimidyl(4-azidophenyl)-1,3′-dithiopropionate (sulfo-sADP), sulfosuccinimidyl 4-(p-azidophenyl)butyrate (sulfo-sAPB), sulfosuccinimidyl 2-(7-azido-4-methylcoumarin-3-acetamide)ethyl-1,3′-dithiopropionate (sAED), sulfosuccinimidyl 7-azido-4-methylcoumain-3-acetate (sulfo-sAMCA), p-nitrophenyl diazopyruvate (pNPDP), p-nitrophenyl-2-diazo-3,3,3-trifluoropropionate (PNP-DTP), sulfhydryl-reactive and photoreactive cross-linkers such asl-(p-Azidosalicylamido)-4-(iodoacetamido)butane (AsIB), N-[4-(p-azidosalicylamido)butyl]-3′-(2′-pyridyldithio)propionamide (APDP), benzophenone-4-iodoacetamide, benzophenone-4-maleimide carbonyl-reactive and photoreactive cross-linkers such as p-azidobenzoyl hydrazide (ABH), carboxylate-reactive and photoreactive cross-linkers such as 4-(p-azidosalicylamido)butylamine (AsBA), and arginine-reactive and photoreactive cross-linkers such as p-azidophenyl glyoxal (APG). 
     In some instances, the linker comprises a reactive functional group. In some cases, the reactive functional group comprises a nucleophilic group that is reactive to an electrophilic group present on a binding moiety. Exemplary electrophilic groups include carbonyl groups-such as aldehyde, ketone, carboxylic acid, ester, amide, enone, acyl halide or acid anhydride. In some embodiments, the reactive functional group is aldehyde. Exemplary nucleophilic groups include hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxylate, and arylhydrazide. 
     In some embodiments, the linker comprises a maleimide goup. In some instances, the maleimide group is also referred to as a maleimide spacer. In some instances, the maleimide group further encompasses a caproic acid, forming maleimidocaproyl (mc). In some cases, the linker comprises maleimidocaproyl (mc). In some cases, the linker is maleimidocaproyl (mc). In other instances, the maleimide group comprises a maleimidomethyl group, such as succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sMCC) or sulfosuccinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (sulfo-sMCC) described above. 
     In some embodiments, the maleimide group is a self-stablizing maleimide. In some instances, the self-stablizing maleimide utilizes diaminopropionic acid (DPR) to incorporate a basic amino group adjacent to the maleimide to provide intramolecular catalysis of tiosuccinimide ring hydrolysis, thereby eliminating maleimide from undergoing an elimination reaction through a retro-Michael reaction. In some instances, the self-stabilizing maleimide is a maleimide group described in Lyon, et al., “Self-hydrolyzing maleimides improve the stability and pharmacological properties of antibody-drug conjugates,”  Nat. Biotechnol.  32(10): 1059-1062 (2014). In some instances, the linker comprises a self-stablizing maleimide. In some instances, the linker is a self-stablizing maleimide. 
     In some embodiments, the linker comprises a peptide moiety. In some instances, the peptide moiety comprises at least 2, 3, 4, 5, or 6 more amino acid residues. In some instances, the peptide moiety comprises at most 2, 3, 4, 5, 6, 7, or 8 amino acid residues. In some instances, the peptide moiety comprises about 2, about 3, about 4, about 5, or about 6 amino acid residues. In some instances, the peptide moiety is a cleavable peptide moiety (e.g., either enzymatically or chemically). In some instances, the peptide moiety is a non-cleavable peptide moiety. In some instances, the peptide moiety comprises Val-Cit (valine-citrulline), Gly-Gly-Phe-Gly, Phe-Lys, Val-Lys, Gly-Phe-Lys, Phe-Phe-Lys, Ala-Lys, Val-Arg, Phe-Cit, Phe-Arg, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Ala, Ala-Leu-Ala-Leu, or Gly-Phe-Leu-Gly. In some instances, the linker comprises a peptide moiety such as: Val-Cit (valine-citrulline), Gly-Gly-Phe-Gly, Phe-Lys, Val-Lys, Gly-Phe-Lys, Phe-Phe-Lys, Ala-Lys, Val-Arg, Phe-Cit, Phe-Arg, Leu-Cit, Ile-Cit, Trp-Cit, Phe-Ala, Ala-Leu-Ala-Leu, or Gly-Phe-Leu-Gly. In some cases, the linker comprises Val-Cit. In some cases, the linker is Val-Cit. 
     In some embodiments, the linker comprises a benzoic acid group, or its derivatives thereof. In some instances, the benzoic acid group or its derivatives thereof comprise paraaminobenzoic acid (PABA). In some instances, the benzoic acid group or its derivatives thereof comprise gamma-aminobutyric acid (GABA). 
     In some embodiments, the linker comprises one or more of a maleimide group, a peptide moiety, and/or a benzoic acid group, in any combination. In some embodiments, the linker comprises a combination of a maleimide group, a peptide moiety, and/or a benzoic acid group. In some instances, the maleimide group is maleimidocaproyl (mc). In some instances, the peptide group is val-cit. In some instances, the benzoic acid group is PABA. In some instances, the linker comprises a mc-val-cit group. In some cases, the linker comprises a val-cit-PABA group. In additional cases, the linker comprises a mc-val-cit-PABA group. 
     In some embodiments, the linker is a self-immolative linker or a self-elimination linker. In some cases, the linker is a self-immolative linker. In other cases, the linker is a self-elimination linker (e.g., a cyclization self-elimination linker). In some instances, the linker comprises a linker described in U.S. Pat. No. 9,089,614 or PCT Publication No. WO2015038426. 
     In some embodiments, the linker is a dendritic type linker. In some instances, the dendritic type linker comprises a branching, multifunctional linker moiety. In some instances, the dendritic type linker is used to increase the molar ratio of polynucleotide B to the binding moiety A. In some instances, the dendritic type linker comprises PAMAM dendrimers. 
     In some embodiments, the linker is a traceless linker or a linker in which after cleavage does not leave behind a linker moiety (e.g., an atom or a linker group) to a binding moiety A, a polynucleotide B, a polymer C, or an endosomolytic moiety D. Exemplary traceless linkers include, but are not limited to, germanium linkers, silicium linkers, sulfur linkers, selenium linkers, nitrogen linkers, phosphorus linkers, boron linkers, chromium linkers, or phenylhydrazide linker. In some cases, the linker is a traceless aryl-triazene linker as described in Hejesen, et al., “A traceless aryl-triazene linker for DNA-directed chemistry,”  Org Biomol Chem  11(15): 2493-2497 (2013). In some instances, the linker is a traceless linker described in Blaney, et al., “Traceless solid-phase organic synthesis,”  Chem. Rev.  102: 2607-2024 (2002). In some instances, a linker is a traceless linker as described in U.S. Pat. No. 6,821,783. 
     In some instances, the linker is a linker described in U.S. Pat. Nos. 6,884,869; 7,498,298; 8,288,352; 8,609,105; or 8,697,688; U.S. Patent Publication Nos. 2014/0127239; 2013/028919; 2014/286970; 2013/0309256; 2015/037360; or 2014/0294851; or PCT Publication Nos. WO2015057699; WO2014080251; WO2014197854; WO2014145090; or WO2014177042. 
     In some embodiments, X, Y, and L are independently a bond or a linker. In some instances, X, Y, and L are independently a bond. In some cases, X, Y, and L are independently a linker. 
     In some instances, X is a bond or a linker. In some instances, X is a bond. In some instances, X is a linker. In some instances, the linker is a C 1 -C 6  alkyl group. In some cases, X is a C 1 -C 6  alkyl group, such as for example, a C 5 , C 4 , C 3 , C 2 , or C 1  alkyl group. In some cases, the C 1 -C 6  alkyl group is an unsubstituted C 1 -C 6  alkyl group. As used in the context of a linker, and in particular in the context of X, alkyl means a saturated straight or branched hydrocarbon radical containing up to six carbon atoms. In some instances, X is a non-polymeric linker. In some instances, X includes a homobifunctional linker or a heterobifunctional linker described supra. In some cases, X includes a heterobifunctional linker. In some cases, X includes sMCC. In other instances, X includes a heterobifunctional linker optionally conjugated to a C 1 -C 6  alkyl group. In other instances, X includes sMCC optionally conjugated to a C 1 -C 5  alkyl group. In additional instances, X does not include a homobifunctional linker or a heterobifunctional linker described supra. 
     In some instances, Y is a bond or a linker. In some instances, Y is a bond. In other cases, Y is a linker. In some embodiments, Y is a C 1 -C 6  alkyl group. In some instances, Y is a homobifunctional linker or a heterobifunctional linker described supra. In some instances, Y is a homobifunctional linker described supra. In some instances, Y is a heterobifunctional linker described supra. In some instances, Y comprises a maleimide group, such as maleimidocaproyl (mc) or a self-stabilizing maleimide group described above. In some instances, Y comprises a peptide moiety, such as Val-Cit. In some instances, Y comprises a benzoic acid group, such as PABA. In additional instances, Y comprises a combination of a maleimide group, a peptide moiety, and/or a benzoic acid group. In additional instances, Y comprises a mc group. In additional instances, Y comprises a mc-val-cit group. In additional instances, Y comprises a val-cit-PABA group. In additional instances, Y comprises a mc-val-cit-PABA group. 
     In some instances, L is a bond or a linker. In some cases, L is a bond. In other cases, L is a linker. In some embodiments, L is a C 1 -C 6  alkyl group. In some instances, L is a homobifunctional linker or a heterobifunctional linker described supra. In some instances, L is a homobifunctional linker described supra. In some instances, L is a heterobifunctional linker described supra. In some instances, L comprises a maleimide group, such as maleimidocaproyl (mc) or a self-stabilizing maleimide group described above. In some instances, L comprises a peptide moiety, such as Val-Cit. In some instances, L comprises a benzoic acid group, such as PABA. In additional instances, L comprises a combination of a maleimide group, a peptide moiety, and/or a benzoic acid group. In additional instances, L comprises a mc group. In additional instances, L comprises a mc-val-cit group. In additional instances, L comprises a val-cit-PABA group. In additional instances, L comprises a mc-val-cit-PABA group. 
     In some embodiments, X 1  and X 2  are each independently a bond or a non-polymeric linker. In some instances, X 1  and X 2  are each independently a bond. In some cases, X 1  and X 2  are each independently a non-polymeric linker. 
     In some instances, X 1  is a bond or a non-polymeric linker. In some instances, X 1  is a bond. In some instances, X 1  is a non-polymeric linker. In some instances, the linker is a C 1 -C 6  alkyl group. In some cases, X 1  is a C 1 -C 6  alkyl group, such as for example, a C 5 , C 4 , C 3 , C 2 , or C 1  alkyl group. In some cases, the C 1 -C 6  alkyl group is an unsubstituted C 1 -C 6  alkyl group. As used in the context of a linker, and in particular in the context of X 1 , alkyl means a saturated straight or branched hydrocarbon radical containing up to six carbon atoms. In some instances, X 1  includes a homobifunctional linker or a heterobifunctional linker described supra. In some cases, X 1  includes a heterobifunctional linker. In some cases, X 1  includes sMCC. In other instances, X 1  includes a heterobifunctional linker optionally conjugated to a C 1 -C 6  alkyl group. In other instances, X 1  includes sMCC optionally conjugated to a C 1 -C 6  alkyl group. In additional instances, X 1  does not include a homobifunctional linker or a heterobifunctional linker described supra. 
     In some instances, X 2  is a bond or a linker. In some instances, X 2  is a bond. In other cases, X 2  is a linker. In additional cases, X 2  is a non-polymeric linker. In some embodiments, X 2  is a C 1 -C 6  alkyl group. In some instances, X 2  is a homobifunctional linker or a heterobifunctional linker described supra. In some instances, X 2  is a homobifunctional linker described supra. In some instances, X 2  is a heterobifunctional linker described supra. In some instances, X 2  comprises a maleimide group, such as maleimidocaproyl (mc) or a self-stabilizing maleimide group described above. In some instances, X 2  comprises a peptide moiety, such as Val-Cit. In some instances, X 2  comprises a benzoic acid group, such as PABA. In additional instances, X 2  comprises a combination of a maleimide group, a peptide moiety, and/or a benzoic acid group. In additional instances, X 2  comprises a mc group. In additional instances, X 2  comprises a mc-val-cit group. In additional instances, X 2  comprises a val-cit-PABA group. In additional instances, X 2  comprises a mc-val-cit-PABA group. 
     Pharmaceutical Formulation 
     In some embodiments, the pharmaceutical formulations described herein are administered to a subject by multiple administration routes, including but not limited to, parenteral (e.g., intravenous, subcutaneous, intramuscular), oral, intranasal, buccal, rectal, or transdermal administration routes. In some instances, the pharmaceutical composition describe herein is formulated for parenteral (e.g., intravenous, subcutaneous, intramuscular, intra-arterial, intraperitoneal, intrathecal, intracerebral, intracerebroventricular, or intracranial) administration. In other instances, the pharmaceutical composition describe herein is formulated for oral administration. In still other instances, the pharmaceutical composition describe herein is formulated for intranasal administration. 
     In some embodiments, the pharmaceutical formulations include, but are not limited to, aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations (e.g., nanoparticle formulations), and mixed immediate and controlled release formulations. 
     In some instances, the pharmaceutical formulation includes multiparticulate formulations. In some instances, the pharmaceutical formulation includes nanoparticle formulations. In some instances, nanoparticles comprise cMAP, cyclodextrin, or lipids. In some cases, nanoparticles comprise solid lipid nanoparticles, polymeric nanoparticles, self-emulsifying nanoparticles, liposomes, microemulsions, or micellar solutions. Additional exemplary nanoparticles include, but are not limited to, paramagnetic nanoparticles, superparamagnetic nanoparticles, metal nanoparticles, fullerene-like materials, inorganic nanotubes, dendrimers (such as with covalently attached metal chelates), nanofibers, nanohorns, nano-onions, nanorods, nanoropes and quantum dots. In some instances, a nanoparticle is a metal nanoparticle, e.g., a nanoparticle of scandium, titanium, vanadium, chromium, manganese, iron, cobalt, nickel, copper, zinc, yttrium, zirconium, niobium, molybdenum, ruthenium, rhodium, palladium, silver, cadmium, hafnium, tantalum, tungsten, rhenium, osmium, iridium, platinum, gold, gadolinium, aluminum, gallium, indium, tin, thallium, lead, bismuth, magnesium, calcium, strontium, barium, lithium, sodium, potassium, boron, silicon, phosphorus, germanium, arsenic, antimony, and combinations, alloys or oxides thereof. 
     In some instances, a nanoparticle includes a core or a core and a shell, as in a core-shell nanoparticle. 
     In some instances, a nanoparticle is further coated with molecules for attachment of functional elements (e.g., with one or more of a polynucleic acid molecule or binding moiety described herein). In some instances, a coating comprises chondroitin sulfate, dextran sulfate, carboxymethyl dextran, alginic acid, pectin, carragheenan, fucoidan, agaropectin, porphyran, karaya gum, gellan gum, xanthan gum, hyaluronic acids, glucosamine, galactosamine, chitin (or chitosan), polyglutamic acid, polyaspartic acid, lysozyme, cytochrome C, ribonuclease, trypsinogen, chymotrypsinogen, a-chymotrypsin, polylysine, polyarginine, histone, protamine, ovalbumin or dextrin or cyclodextrin. In some instances, a nanoparticle comprises a graphene-coated nanoparticle. 
     In some cases, a nanoparticle has at least one dimension of less than about 500 nm, 400 nm, 300 nm, 200 nm, or 100 nm. 
     In some instances, the nanoparticle formulation comprises paramagnetic nanoparticles, superparamagnetic nanoparticles, metal nanoparticles, fullerene-like materials, inorganic nanotubes, dendrimers (such as with covalently attached metal chelates), nanofibers, nanohorns, nano-onions, nanorods, nanoropes or quantum dots. In some instances, a polynucleic acid molecule or a binding moiety described herein is conjugated either directly or indirectly to the nanoparticle. In some instances, at least 1, 5, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more polynucleic acid molecules or binding moieties described herein are conjugated either directly or indirectly to a nanoparticle. 
     In some embodiments, the pharmaceutical formulation comprise a delivery vector, e.g., a recombinant vector, the delivery of the polynucleic acid molecule into cells. In some instances, the recombinant vector is DNA plasmid. In other instances, the recombinant vector is a viral vector. Exemplary viral vectors include vectors derived from adeno-associated virus, retrovirus, adenovirus, or alphavirus. In some instances, the recombinant vectors capable of expressing the polynucleic acid molecules provide stable expression in target cells. In additional instances, viral vectors are used that provide for transient expression of polynucleic acid molecules. 
     In some embodiments, the pharmaceutical formulations include a carrier or carrier materials selected on the basis of compatibility with the composition disclosed herein, and the release profile properties of the desired dosage form. Exemplary carrier materials include, e.g., binders, suspending agents, disintegration agents, filling agents, surfactants, solubilizers, stabilizers, lubricants, wetting agents, diluents, and the like. Pharmaceutically compatible carrier materials include, but are not limited to, acacia, gelatin, colloidal silicon dioxide, calcium glycerophosphate, calcium lactate, maltodextrin, glycerine, magnesium silicate, polyvinylpyrrollidone (PVP), cholesterol, cholesterol esters, sodium caseinate, soy lecithin, taurocholic acid, phosphotidylcholine, sodium chloride, tricalcium phosphate, dipotassium phosphate, cellulose and cellulose conjugates, sugars sodium stearoyl lactylate, carrageenan, monoglyceride, diglyceride, pregelatinized starch, and the like. See, e.g., Remington: The Science and Practice ofPharmacy, Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E.,  Remington&#39;s Pharmaceutical Sciences , Mack Publishing Co., Easton, Pa. 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,  Marcel Decker , New York, N.Y., 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &amp; Wilkins 1999). 
     In some instances, the pharmaceutical formulations further include pH adjusting agents or buffering agents which include acids such as acetic, boric, citric, lactic, phosphoric and hydrochloric acids; bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate and tris-hydroxymethylaminomethane; and buffers such as citrate/dextrose, sodium bicarbonate and ammonium chloride. Such acids, bases and buffers are included in an amount required to maintain pH of the composition in an acceptable range. 
     In some instances, the pharmaceutical formulation includes one or more salts in an amount required to bring osmolality of the composition into an acceptable range. Such salts include those having sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anions; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite and ammonium sulfate. 
     In some instances, the pharmaceutical formulations further include diluent which are used to stabilize compounds because they provide a more stable environment. Salts dissolved in buffered solutions (which also provide pH control or maintenance) are utilized as diluents in the art, including, but not limited to a phosphate buffered saline solution. In certain instances, diluents increase bulk of the composition to facilitate compression or create sufficient bulk for homogenous blend for capsule filling. Such compounds include e.g., lactose, starch, mannitol, sorbitol, dextrose, microcrystalline cellulose such as Avicel®; dibasic calcium phosphate, dicalcium phosphate dihydrate; tricalcium phosphate, calcium phosphate; anhydrous lactose, spray-dried lactose; pregelatinized starch, compressible sugar, such as Di-Pac® (Amstar); mannitol, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose acetate stearate, sucrose-based diluents, confectioner&#39;s sugar; monobasic calcium sulfate monohydrate, calcium sulfate dihydrate; calcium lactate trihydrate, dextrates; hydrolyzed cereal solids, amylose; powdered cellulose, calcium carbonate; glycine, kaolin; mannitol, sodium chloride; inositol, bentonite, and the like. 
     In some cases, the pharmaceutical formulations include disintegration agents or disintegrants to facilitate the breakup or disintegration of a substance. The term “disintegrate” include both the dissolution and dispersion of the dosage form when contacted with gastrointestinal fluid. Examples of disintegration agents include a starch, e.g., a natural starch such as corn starch or potato starch, a pregelatinized starch such as National 1551 or Amijel®, or sodium starch glycolate such as Promogel® or Explotab®, a cellulose such as a wood product, methylcrystalline cellulose, e.g., Avicel®, Avicel® PH101, Avicel® PH102, Avicel® PH105, Elcema® P100, Emcocel®, Vivacel®, Ming Tia®, and Solka-Floc®, methylcellulose, croscarmellose, or a cross-linked cellulose, such as cross-linked sodium carboxymethylcellulose (Ac-Di-Sol®), cross-linked carboxymethylcellulose, or cross-linked croscarmellose, a cross-linked starch such as sodium starch glycolate, a cross-linked polymer such as crospovidone, a cross-linked polyvinylpyrrolidone, alginate such as alginic acid or a salt of alginic acid such as sodium alginate, a clay such as Veegum® HV (magnesium aluminum silicate), a gum such as agar, guar, locust bean, Karaya, pectin, or tragacanth, sodium starch glycolate, bentonite, a natural sponge, a surfactant, a resin such as a cation-exchange resin, citrus pulp, sodium lauryl sulfate, sodium lauryl sulfate in combination starch, and the like. 
     In some instances, the pharmaceutical formulations include filling agents such as lactose, calcium carbonate, calcium phosphate, dibasic calcium phosphate, calcium sulfate, microcrystalline cellulose, cellulose powder, dextrose, dextrates, dextran, starches, pregelatinized starch, sucrose, xylitol, lactitol, mannitol, sorbitol, sodium chloride, polyethylene glycol, and the like. 
     Lubricants and glidants are also optionally included in the pharmaceutical formulations described herein for preventing, reducing or inhibiting adhesion or friction of materials. Exemplary lubricants include, e.g., stearic acid, calcium hydroxide, talc, sodium stearyl fumerate, a hydrocarbon such as mineral oil, or hydrogenated vegetable oil such as hydrogenated soybean oil (Sterotex®), higher fatty acids and their alkali-metal and alkaline earth metal salts, such as aluminum, calcium, magnesium, zinc, stearic acid, sodium stearates, glycerol, talc, waxes, Stearowet®, boric acid, sodium benzoate, sodium acetate, sodium chloride, leucine, a polyethylene glycol (e.g., PEG-4000) or a methoxypolyethylene glycol such as Carbowax™, sodium oleate, sodium benzoate, glyceryl behenate, polyethylene glycol, magnesium or sodium lauryl sulfate, colloidal silica such as Syloid™, Cab-O-Sil®, a starch such as corn starch, silicone oil, a surfactant, and the like. 
     Plasticizers include compounds used to soften the microencapsulation material or film coatings to make them less brittle. Suitable plasticizers include, e.g., polyethylene glycols such as PEG 300, PEG 400, PEG 600, PEG 1450, PEG 3350, and PEG 800, stearic acid, propylene glycol, oleic acid, triethyl cellulose and triacetin. Plasticizers also function as dispersing agents or wetting agents. 
     Solubilizers include compounds such as triacetin, triethylcitrate, ethyl oleate, ethyl caprylate, sodium lauryl sulfate, sodium doccusate, vitamin E TPGS, dimethylacetamide, N-methylpyrrolidone, N-hydroxyethylpyrrolidone, polyvinylpyrrolidone, hydroxypropylmethyl cellulose, hydroxypropyl cyclodextrins, ethanol, n-butanol, isopropyl alcohol, cholesterol, bile salts, polyethylene glycol 200-600, glycofurol, transcutol, propylene glycol, and dimethyl isosorbide and the like. 
     Stabilizers include compounds such as any antioxidation agents, buffers, acids, preservatives and the like. 
     Suspending agents include compounds such as polyvinylpyrrolidone, e.g., polyvinylpyrrolidone K12, polyvinylpyrrolidone K17, polyvinylpyrrolidone K25, or polyvinylpyrrolidone K30, vinyl pyrrolidone/vinyl acetate copolymer (S630), polyethylene glycol, e.g., the polyethylene glycol has a molecular weight of about 300 to about 6000, or about 3350 to about 4000, or about 7000 to about 5400, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, hydroxymethylcellulose acetate stearate, polysorbate-80, hydroxyethylcellulose, sodium alginate, gums, such as, e.g., gum tragacanth and gum acacia, guar gum, xanthans, including xanthan gum, sugars, cellulosics, such as, e.g., sodium carboxymethylcellulose, methylcellulose, sodium carboxymethylcellulose, hydroxypropylmethylcellulose, hydroxyethylcellulose, polysorbate-80, sodium alginate, polyethoxylated sorbitan monolaurate, polyethoxylated sorbitan monolaurate, povidone and the like. 
     Surfactants include compounds such as sodium lauryl sulfate, sodium docusate, Tween 60 or 80, triacetin, vitamin E TPGS, sorbitan monooleate, polyoxyethylene sorbitan monooleate, polysorbates, polaxomers, bile salts, glyceryl monostearate, copolymers of ethylene oxide and propylene oxide, e.g., Pluronic® (BASF), and the like. Additional surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, e.g., polyoxyethylene (60) hydrogenated castor oil; and polyoxyethylene alkylethers and alkylphenyl ethers, e.g., octoxynol 10, octoxynol 40. Sometimes, surfactants is included to enhance physical stability or for other purposes. 
     Viscosity enhancing agents include, e.g., methyl cellulose, xanthan gum, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hydroxypropylmethyl cellulose acetate stearate, hydroxypropylmethyl cellulose phthalate, carbomer, polyvinyl alcohol, alginates, acacia, chitosans and combinations thereof. 
     Wetting agents include compounds such as oleic acid, glyceryl monostearate, sorbitan monooleate, sorbitan monolaurate, triethanolamine oleate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monolaurate, sodium docusate, sodium oleate, sodium lauryl sulfate, sodium doccusate, triacetin, Tween 80, vitamin E TPGS, ammonium salts and the like. 
     Therapeutic Regimens 
     In some embodiments, the pharmaceutical compositions described herein are administered for therapeutic applications. In some embodiments, the pharmaceutical composition is administered once per day, twice per day, three times per day or more. The pharmaceutical composition is administered daily, every day, every alternate day, five days a week, once a week, every other week, two weeks per month, three weeks per month, once a month, twice a month, three times per month, or more. The pharmaceutical composition is administered for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 18 months, 2 years, 3 years, or more. 
     In some embodiments, one or more pharmaceutical compositions are administered simultaneously, sequentially, or at an interval period of time. In some embodiments, one or more pharmaceutical compositions are administered simultaneously. In some cases, one or more pharmaceutical compositions are administered sequentially. In additional cases, one or more pharmaceutical compositions are administered at an interval period of time (e.g., the first administration of a first pharmaceutical composition is on day one followed by an interval of at least 1, 2, 3, 4, 5, or more days prior to the administration of at least a second pharmaceutical composition). 
     In some embodiments, two or more different pharmaceutical compositions are coadministered. In some instances, the two or more different pharmaceutical compositions are coadministered simultaneously. In some cases, the two or more different pharmaceutical compositions are coadministered sequentially without a gap of time between administrations. In other cases, the two or more different pharmaceutical compositions are coadministered sequentially with a gap of about 0.5 hour, 1 hour, 2 hour, 3 hour, 12 hours, 1 day, 2 days, or more between administrations. 
     In the case wherein the patient&#39;s status does improve, upon the doctor&#39;s discretion the administration of the composition is given continuously; alternatively, the dose of the composition being administered is temporarily reduced or temporarily suspended for a certain length of time (i.e., a “drug holiday”). In some instances, the length of the drug holiday varies between 2 days and 1 year, including by way of example only, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50 days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250 days, 280 days, 300 days, 320 days, 350 days, or 365 days. The dose reduction during a drug holiday is from 10%-100%, including, by way of example only, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. 
     Once improvement of the patient&#39;s conditions has occurred, a maintenance dose is administered if necessary. Subsequently, the dosage or the frequency of administration, or both, can be reduced, as a function of the symptoms, to a level at which the improved disease, disorder or condition is retained. 
     In some embodiments, the amount of a given agent that correspond to such an amount varies depending upon factors such as the particular compound, the severity of the disease, the identity (e.g., weight) of the subject or host in need of treatment, but nevertheless is routinely determined in a manner known in the art according to the particular circumstances surrounding the case, including, e.g., the specific agent being administered, the route of administration, and the subject or host being treated. In some instances, the desired dose is conveniently presented in a single dose or as divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, for example as two, three, four or more sub-doses per day. 
     The foregoing ranges are merely suggestive, as the number of variables in regard to an individual treatment regime is large, and considerable excursions from these recommended values are not uncommon. Such dosages is altered depending on a number of variables, not limited to the activity of the compound used, the disease or condition to be treated, the mode of administration, the requirements of the individual subject, the severity of the disease or condition being treated, and the judgment of the practitioner. 
     In some embodiments, toxicity and therapeutic efficacy of such therapeutic regimens are determined by standard pharmaceutical procedures in cell cultures or experimental animals, including, but not limited to, the determination of the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between the toxic and therapeutic effects is the therapeutic index and it is expressed as the ratio between LD50 and ED50. Compounds exhibiting high therapeutic indices are preferred. The data obtained from cell culture assays and animal studies are used in formulating a range of dosage for use in human. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with minimal toxicity. The dosage varies within this range depending upon the dosage form employed and the route of administration utilized. 
     Kits/Article of Manufacture 
     Disclosed herein, in certain embodiments, are kits and articles of manufacture for use with one or more of the compositions and methods described herein. Such kits include a carrier, package, or container that is compartmentalized to receive one or more containers such as vials, tubes, and the like, each of the container(s) comprising one of the separate elements to be used in a method described herein. Suitable containers include, for example, bottles, vials, syringes, and test tubes. In one embodiment, the containers are formed from a variety of materials such as glass or plastic. 
     The articles of manufacture provided herein contain packaging materials. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, bags, containers, bottles, and any packaging material suitable for a selected formulation and intended mode of administration and treatment. 
     For example, the container(s) include target nucleic acid molecule described herein. Such kits optionally include an identifying description or label or instructions relating to its use in the methods described herein. 
     A kit typically includes labels listing contents and/or instructions for use, and package inserts with instructions for use. A set of instructions will also typically be included. 
     In one embodiment, a label is on or associated with the container. In one embodiment, a label is on a container when letters, numbers or other characters forming the label are attached, molded or etched into the container itself; a label is associated with a container when it is present within a receptacle or carrier that also holds the container, e.g., as a package insert. In one embodiment, a label is used to indicate that the contents are to be used for a specific therapeutic application. The label also indicates directions for use of the contents, such as in the methods described herein. 
     In certain embodiments, the pharmaceutical compositions are presented in a pack or dispenser device which contains one or more unit dosage forms containing a compound provided herein. The pack, for example, contains metal or plastic foil, such as a blister pack. In one embodiment, the pack or dispenser device is accompanied by instructions for administration. In one embodiment, the pack or dispenser is also accompanied with a notice associated with the container in form prescribed by a governmental agency regulating the manufacture, use, or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the drug for human or veterinary administration. Such notice, for example, is the labeling approved by the U.S. Food and Drug Administration for prescription drugs, or the approved product insert. In one embodiment, compositions containing a compound provided herein formulated in a compatible pharmaceutical carrier are also prepared, placed in an appropriate container, and labeled for treatment of an indicated condition. 
     Certain Terminology 
     Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise. In this application, the use of “or” means “and/or” unless stated otherwise. Furthermore, use of the term “including” as well as other forms, such as “include”, “includes,” and “included,” is not limiting. 
     As used herein, ranges and amounts can be expressed as “about” a particular value or range. About also includes the exact amount. Hence “about 5 μL” means “about 5 μL” and also “5 μL.” Generally, the term “about” includes an amount that would be expected to be within experimental error. 
     The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. 
     As used herein, the terms “individual(s)”, “subject(s)” and “patient(s)” mean any mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human. None of the terms require or are limited to situations characterized by the supervision (e.g. constant or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a physician&#39;s assistant, an orderly or a hospice worker). 
     EXAMPLES 
     These examples are provided for illustrative purposes only and not to limit the scope of the claims provided herein. 
     Example 1. Antisense Oligonucleotide Sequences and Synthesis 
     Phosphorodiamidate morpholino oligomers (PMO), phosphorothioate antisense oligonucleotides (PS ASO), and antisense oligonucleotides (ASOs) were synthesized. 
     The PMO sequence was 5′GGCCAAACCTCGGCTTACCTGAAAT3′ Primary amine (SEQ ID NO: 28) and can be seen in  FIG. 1  with end nucleotides expanded. The PMO contains a C3-NH 2  conjugation handle at the 3′ end of the molecule for conjugation. PMOs were fully assembled on solid phase using standard solid phase synthesis protocols and purified over HPLC. 
     The PS ASO sequence was Amine-C6-GGCCAAACCUCGGCUUACCU (SEQ ID NO: 29) and can be seen in  FIGS. 2A-2B  with end nucleotides expanded. The structure of the PS ASO comprised a phosphate backbone that was 100% phosphorothioate linkages and all the ribose sugars contained a 2′ 2′OMe modification. The PS ASO also contained a C6-NH 2  conjugation handle at the 5′ end of the molecule for conjugation. The PS ASOs were fully assembled on the solid phase using standard solid phase phosphoramidite chemistry and purified over HPLC. 
     ASOs were fully assembled on the solid phase using standard solid phase phosphoramidite chemistry and purified over HPLC. ASOs contained a C6-NH 2  conjugation handle at the 5′ end of the molecule for conjugation. 
     Example 2. Detection of DMD Exon Skipping 
     Methods for Determining DMD Exon 23 Skipping in Differentiated C1C12 Cells 
     Mouse myoblast C2C12 cells were plated at 50,000-100,000/well in 24-well plates in 0.5 mL 10% FBS RPMI 1640 media and incubated at 37° C. with 5% CO 2  overnight. On the second day, cells were switched to differentiation media (2% horse serum RPMI 1640 and 1 μM insulin) and incubated for 3-5 days. Following incubation, samples were added and incubated for 24 hours. After the sample treatment, 1 mL of fresh media (with no compounds) was changed every day for 2 more days. At 72 hours after the start of treatments, cells were harvested. RNAs were isolated using InviTrap RNA Cell HTS 96 Kit (B-Bridge International #7061300400) and reverse transcribed using High Capacity cDNA Reverse transcription Kit (ThermoFisher #4368813). PCR reactions were performed using DreamTaq™ PCR Mastermix (ThermoFisher # K1072). The primary PCR used primers in exon 20 (Ex20F 5′-CAGAATTCTGCCAATTGCTGAG) (SEQ ID NO: 30) and exon 26 (Ex26R 5′-TTCTTCAGCTTGTGTCATCC) (SEQ ID NO: 31) to amplify both skipped and unskipped molecules using the protocol in Table 2. 
     
       
         
           
               
               
             
               
                 TABLE 2 
               
               
                   
               
               
                   
                 PCR Protocol 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 Hot Start 
                 95° C. for 2 minutes 
               
               
                   
                 Denaturation 
                 95° C. for 0.5 minute 
               
               
                   
                 Annealing of primers 
                 50° C. for 0.5 minute 
               
               
                   
                 Primer extension 
                 72° C. for 1 minute 
               
               
                   
                 Final extension 
                 72° C. for 5 minutes 
               
               
                   
                 Number of Cycles 
                 10 
               
               
                   
               
            
           
         
       
     
     For the nested PCR, primary PCR reactions were diluted with water 100×, and 5 μl was used for nested PCR reaction (50 μl total reaction volume). Nested PCR used primers in exon 20 (Ex20F2: 5′-ACCCAGTCTACCACCCTATC) (SEQ ID NO: 32) and exon 25 (Ex25R: 5′-CTCTTTATCTTCTGCCCACCTT) (SEQ ID NO: 33) to amplify both skipped and unskipped molecules using the protocol in Table 3. 
     
       
         
           
               
             
               
                 TABLE 3 
               
               
                   
               
               
                 Nested PCR Protocol 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
            
               
                   
                 Hot Start 
                 95° C. for 2 minutes 
               
               
                   
                 Denaturation 
                 95° C. for 0.5 minute 
               
               
                   
                 Annealing of primers 
                 50° C. for 0.5 minute 
               
               
                   
                 Primer extension 
                 72° C. for 1 minute 
               
               
                   
                 Final extension 
                 72° C. for 5 minutes 
               
               
                   
                 Number of Cycles 
                 35 
               
               
                   
               
            
           
         
       
     
     PCR reactions were analyzed using 4% TAE agarose gels. The wild-type (WT) DMD product had an expected size of 788 base pairs and the skipped DMD A23 of 575 base pairs. 
     Animals 
     All animal studies were conducted following protocols in accordance with the Institutional Animal Care and Use Committee (IACUC) at Explora BioLabs, which adhere to the regulations outlined in the USDA Animal Welfare Act as well as the “Guide for the Care and Use of Laboratory Animals” (National Research Council publication, 8th Ed., revised in 2011). All mice were obtained from either Charles River Laboratories or Harlan Laboratories. 
     In Vivo Mouse Model 
     WT CD-1 mice (4-6 weeks old) were dosed via intravenous (iv) injection with the indicated antisense conjugates (ASCs) and doses. The “naked” PMO or ASO were dosed via intramuscular injection at the indicated doses. After 4, 7, or 14 days, heart and gastrocnemius muscle tissues were harvested and snap-frozen in liquid nitrogen. RNAs were isolated with Trizol and RNeasy Plus 96 Kit (Qiagen, #74192) and reversed transcribed using High Capacity cDNA Reverse transcription Kit (ThermoFisher #4368813). Nested PCR reactions were performed as described. PCR reactions were analyzed in 4% TAE agarose gels which were quantitated by densitometry. 
     To confirm exon 23 skipping in treated mice, DNA fragments were isolated from the 4% agarose gels and sequenced. 
     To quantitatively determine the skipped DMD mRNA copy number, qPCR primer/probe sets were designed to quantify skipped and WT DMD mRNA ( FIG. 3 ). qPCR quantification standards were designed and produced via PCR using designed PCR primers as seen in Table 4. For the qPCR standard for WT and DMD, following PCR a 733 base pair fragment was isolated from the agarose gel. For qPCR standard for skipped DMA, the nested primers were used. 
     The amplification efficiency of the qPCR primer/probes were determined to be within 10% of expected efficiency. qPCR reactions were performed in QuantStudio 7 and Taqman™ PCR Universal Mastermix II (ThermoFisher #4440041) according to manufacturer&#39;s instructions. 
     
       
         
           
               
               
               
               
             
               
                 TABLE 4 
               
               
                   
               
               
                   
                 SEQ 
                   
                   
               
               
                   
                 ID 
                   
                   
               
               
                   
                 NO 
                 Primer/Probe 
                 Sequence 
               
               
                   
               
             
            
               
                 DMD Δ-23, for 
                 34 
                 Forward Primer 
                 5′ GCGCTATCA 
               
               
                 Ex23 skipping 
                   
                   
                 GGAGACAATGAG 
               
               
                   
                 35 
                 Reverse Primer 
                 5′ GTTTTTATGTGA 
               
               
                   
                   
                   
                 TTCTGTAATTTCCC 
               
               
                   
                 36 
                 Probe 
                 5′ CTCTCTGTACCT 
               
               
                   
                   
                   
                 TATCTTAGTGTT 
               
               
                   
               
               
                 DMD Ex22-23, 
                 37 
                 Forward Primer 
                 5′ TGGAGGAGA 
               
               
                 for WT DMD 
                   
                   
                 GACTCGGGAAA 
               
               
                 only 
                 38 
                 Reverse Primer 
                 5′ TTGAAGCCAT 
               
               
                   
                   
                   
                 TTTGTTGCTCTTT 
               
               
                   
                 39 
                 Probe 
                 5′ ACAGGCTCTG 
               
               
                   
                   
                   
                 CAAAGT 
               
               
                   
               
               
                 DMD Ex20-21, 
                 40 
                 Forward Primer 
                 5′ AACAGATGACA 
               
               
                 for All DMD 
                   
                   
                 ACTACTGCCGAAA 
               
               
                   
                 41 
                 Reverse Primer 
                 5′ TTGGCTCTGAT 
               
               
                   
                   
                   
                 AGGGTGGTAGAC 
               
               
                   
                 42 
                 Probe 
                 5′ CTTGTTGAAAA 
               
               
                   
                   
                   
                 CCC 
               
               
                   
               
               
                 qPCR standard 
                 43 
                 Forward Primer 
                 5′ TGAGGGTGTTA 
               
               
                 for WT and all 
                   
                   
                 ATGCTGAAAGTA 
               
               
                 DMD 
                 44 
                 Reverse Primer 
                 5′ CACCAACTGGG 
               
               
                   
                   
                   
                 AGGAAAGTT 
               
               
                   
               
            
           
         
       
     
     Example 3: Conjugate Synthesis 
     Analytical and Purification Methods 
     Analytical and purification methods were performed according to Tables 5-11. 
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Size exclusion chromatography (SEC) methods 
               
            
           
           
               
               
               
               
            
               
                 Size Exclusion 
                   
                   
                   
               
               
                 Chromatography  
                   
                 Mobile 
                   
               
               
                 (SEC) Method 
                 Column 
                 Phase 
                 Flow Rate 
               
               
                   
               
               
                 method 1 
                 TOSOH Biosciences, 
                 150 mM  
                 1.0 mL/  
               
               
                   
                 TSKgelG3000SW XL, 
                 phosphate 
                 minute for 
               
               
                   
                 7.8 × 300 mm, 5 μM 
                 buffer 
                 20 minutes 
               
               
                 method 2 
                 TOSOH Biosciences, 
                 PBS pH 7.4 
                 1.0 mL/  
               
               
                   
                 TSKgelG3000SW,  
                   
                 minute for 
               
               
                   
                 21.5 × 600 mm, 5 μM 
                   
                 180 minutes 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 6 
               
             
            
               
                   
               
               
                 Hydrophobic interaction chromatography (HIC) method 1 
               
            
           
           
               
               
               
            
               
                   
                   
                 Gradient 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Column 
                   
                   
               
               
                 Column 
                 Solvent 
                 Volume 
                 % A 
                 % B 
               
               
                   
               
               
                 GE, HiScreen  
                 Solvent A: 50 mM phosphate buffer,  
                  1.00 
                 95 
                  5 
               
               
                 Butyl 
                 0.8 M Ammonium Sulfate, pH 7.0 
                 30 
                  0 
                 100 
               
               
                 HP, 4.7 mL 
                 Solvent B: 80% 50 mM phosphate  
                  5 
                  0 
                 100 
               
               
                   
                 buffer, 20% IPA, pH 7.0 
                   
                   
                   
               
               
                   
                 Flow Rate: 1.0 mL/minute 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 7 
               
             
            
               
                   
               
               
                 Hydrophobic interaction chromatography (HIC) method 2 
               
            
           
           
               
               
               
            
               
                   
                   
                 Gradient 
               
            
           
           
               
               
               
               
               
            
               
                 Column 
                 Solvent 
                 Time 
                 % A 
                 % B 
               
               
                   
               
               
                 Thermo Scientific, 
                 Solvent A: 100 mM  
                  0.00 
                 100 
                  0 
               
               
                 MAbPac HIC-20, 
                 phosphate buffer,  
                  2.00 
                 100 
                  0 
               
               
                 4.6 mm ID × 10 cm, 
                 1.8 M Ammonium 
                   
                   
                   
               
               
                 5 um 
                 Sulfate, pH 7.0 
                   
                   
                   
               
               
                   
                 Solvent B: 80% 100 mM  
                 22.00 
                  0 
                 100 
               
               
                   
                 phosphate buffer, 20%  
                 25.00 
                  0 
                 100 
               
               
                   
                 IPA, pH 7.0 
                 26.00 
                 100 
                  0 
               
               
                   
                 Flow Rate: 0.7 mL/minute 
                 30.00 
                 100 
                  0 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 8 
               
             
            
               
                   
               
               
                 Hydrophobic interaction chromatography (HIC) method 3 
               
            
           
           
               
               
               
            
               
                   
                   
                 Gradient 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Column 
                   
                   
               
               
                 Column 
                 Solvent 
                 Volume 
                 % A 
                 % B 
               
               
                   
               
               
                 GE, HiScreen  
                 Solvent A: 50 mM phosphate  
                  1 
                 100 
                  0 
               
               
                 Butyl 
                 buffer, 0.8 M Ammonium  
                 25 
                  0 
                  80 
               
               
                 HP, 4.7 mL 
                 Sulfate, pH 7.0 
                   
                   
                   
               
               
                   
                 Solvent B: 80% 50 mM  
                  1 
                  0 
                 100 
               
               
                   
                 phosphate buffer, 
                   
                   
                   
               
               
                   
                 20% IPA, pH 7.0 
                  2 
                  0 
                 100 
               
               
                   
                 Flow Rate: 1.0 mL/minute 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 9 
               
             
            
               
                   
               
               
                 Hydrophobic interaction chromatography (HIC) method 4 
               
            
           
           
               
               
               
            
               
                   
                   
                 Gradient 
               
            
           
           
               
               
               
               
               
            
               
                 Column 
                 Solvent 
                 Time 
                 % A 
                 % B 
               
               
                   
               
               
                 Thermo Scientific, 
                 Solvent A: 100 mM phosphate  
                  0.00 
                 100 
                  0 
               
               
                 MAbPac HIC-20,  
                 buffer, 1.8 M Ammonium 
                  5.00 
                 100 
                  0 
               
               
                 4.6 mm ID × 10 cm, 
                 Sulfate, pH 7.0 
                   
                   
                   
               
               
                 5 um 
                 Solvent B: 80% 100 mM  
                 20.00 
                  0 
                 100 
               
               
                   
                 phosphate buffer, 
                 25.00 
                  0 
                 100 
               
               
                   
                 20% IPA, pH 7.0 
                 26.00 
                 100 
                  0 
               
               
                   
                 Flow Rate: 0.5 mL/minute 
                 30.00 
                 100 
                  0 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 10 
               
             
            
               
                   
               
               
                 Strong anion exchange chromatography (SAX) method 1 
               
            
           
           
               
               
               
            
               
                   
                   
                 Gradient 
               
            
           
           
               
               
               
               
               
            
               
                   
                   
                 Column 
                   
                   
               
               
                 Column 
                 Solvent 
                 Volume 
                 % A 
                 % B 
               
               
                   
               
               
                 Tosoh Bioscience, 
                 Solvent A: 20 mM TRIS  
                  0.5 
                 100 
                  0 
               
               
                 TSKGel SuperQ- 
                 buffer, pH 8.0; 
                   
                   
                   
               
               
                 5PW, 21.5 mm  
                 Solvent B: 20 mM TRIS,  
                  0.5 
                  80 
                  20 
               
               
                 ID × 15 cm,  
                 1.5 M NaCl, pH 8.0 
                 17 
                  20 
                  80 
               
               
                 13 um 
                 Flow Rate: 6.0 mL/ 
                  0.5 
                  0 
                 100 
               
               
                   
                 minute 
                  0.5 
                  0 
                 100 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 11 
               
             
            
               
                   
               
               
                 Strong anion exchange chromatography (SAX) method 2 
               
            
           
           
               
               
               
            
               
                   
                   
                 Gradient 
               
            
           
           
               
               
               
               
               
            
               
                 Column 
                 Solvent 
                 Time 
                 % A 
                 % B 
               
               
                   
               
               
                 Thermo Scientific, 
                 Solvent A: 80% 10 mM TRIS  
                  0.0 
                 90 
                  10 
               
               
                 ProPac ™ SAX-10, 
                 pH 8, 20% ethanol 
                  3.00 
                 90 
                  10 
               
               
                 Bio LC ™ 4 × 250 
                 Solvent B: 80% 10 mM TRIS  
                 17.00 
                  0 
                 100 
               
               
                 mm 
                 pH 8, 20% ethanol,  
                 21.00 
                  0 
                 100 
               
               
                   
                 1.5 M NaCl 
                 22.00 
                 90 
                  10 
               
               
                   
                 Flow Rate: 0.75 mL/minute 
                 25.00 
                 90 
                  10 
               
               
                   
               
            
           
         
       
     
     Anti-Transferrin Receptor Antibody 
     Anti-mouse transferrin receptor antibody or anti-CD71 mAb that was used was a rat IgG2a subclass monoclonal antibody that binds mouse CD71 or mouse transferrin receptor 1 (mTfR1). The antibody was produced by BioXcell and it is commercially available (Catalog # BE0175). 
     Anti-CD71 Antibody Morpholino Antisense Oligonucleotide Conjugate (Anti-CD71 mAb-PMO) 
     Anti-CD71 mAb-PMO Conjugation 
     Anti-CD71 antibody (10 mg/mL) in borate buffer (25 mM sodium tetraborate, 25 mM NaCl, 1 mM Diethylene triamine pentaacetic acid, pH 8.0) was reduced by adding 4 equivalents of tris(2-carboxyethyl)phosphine (TCEP) in water and incubating at 37° C. for 4 hours. 4(N-Maleimidomethyl)cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) was coupled to the primary amine on the 3′ end of the phosphorodiamidate morpholino oligomer (PMO) by incubating the PMO (50 mg/mL) in DMSO with 10 equivalents of SMCC (10 mg/mL) in DMSO for one hour. Unconjugated SMCC was removed by ultrafiltration using Amicon Ultra-15 centrifugal filter units with a MWCO of 3 kDa. The PMO-SMCC was washed three times with acetate buffer (10 mM sodium acetate, pH 6.0) and used immediately. The reduced antibody was mixed with 2.25 equivalents of PMO-SMCC and incubated overnight at 4° C. The pH of the reaction mixture was then reduced to 7.5, and 8 equivalents of N-Ethylmaleimide was added to the mixture at room temperature for 30 minutes to quench unreacted cysteines. Analysis of the reaction mixture by hydrophobic interaction chromatography (HIC) method 2 showed antibody-PMO conjugates along with unreacted antibody and PMO ( FIG. 4 ).  FIG. 4  shows a chromatogram of anti-CD71 mAb-PMO reaction mixture produced with HIC method 2 showing free antibody peak (1), free PMO (2), DAR 1 (3), DAR 2 (4), DAR 3 (5), DAR&gt;3 (6). “DAR” refers to a drug-to-antibody ratio. The number in parentheses refers to the peak in the chromatogram. 
     Purification 
     The reaction mixture was purified with an AKTA Explorer FPLC using HIC method 1. Fractions containing conjugates with a drug to antibody ratio of one (DAR 1) and two (DAR 2) were combined and concentrated with Amicon Ultra-15 centrifugal filter units with a MWCO of 50 kDa separately from conjugates with a DAR greater than 2. Concentrated conjugates were buffer exchanged with PBS (pH 7.4) using Amicon Ultra-15 centrifugal filter units prior to analysis. 
     Analysis of the Purified Conjugate 
     The isolated conjugates were characterized by size exclusion chromatography (SEC) and HIC. SEC method 1 was used to confirm the absence of high molecular weight aggregates and unconjugated PMOs ( FIGS. 5A-5C ).  FIG. 5A  shows a chromatogram of anti-CD71 mAb produced using SEC method 1.  FIG. 5B  shows a chromatogram of anti-CD71 mAb-PMO DAR 1,2 produced using SEC method 1.  FIG. 5C  shows a chromatogram of anti-CD71 mAb-PMO DAR greater than 2 produced using SEC method 1. “DAR” refers to a drug-to-antibody ratio. 
     The purity of the conjugate was assessed by analytical HPLC using HIC method 2 ( FIGS. 6A-6C ).  FIG. 6A  shows a chromatogram of anti-CD71 mAb produced using HIC method 2.  FIG. 6B  shows a chromatogram of purified anti-CD71 mAb-PMO DAR 1,2 conjugate produced using HIC method 2.  FIG. 6C  shows a chromatogram of purified anti-CD71 mAb-PMO DAR&gt;2 conjugate produced using HIC method 2. The 260/280 nm UV absorbance ratio of each sample was compared to a standard curve of known ratios of PMO and antibody to confirm DAR. The DAR 1,2 sample had an average DAR of ˜1.6 while the DAR greater than 2 sample had an average DAR of ˜3.7. “DAR” refers to a drug-to-antibody ratio. 
     Anti-CD71 Fab Morpholino Antisense Oligonucleotide Conjugate (Anti-CD71 Fab-PMO) 
     Antibody Digestion with Pepsin 
     Anti-CD71 antibody (5 mg/mL) in 20 mM acetate buffer (pH 4.0) was incubated with immobilized pepsin for 3 hours at 37° C. The resin was removed and the reaction mixture was washed with PBS (pH 7.4) using Amicon Ultra-15 centrifugal filter units with a MWCO of 30 kDa. The retentate was collected and purified using size exclusion chromatography (SEC) method 2 to isolate the F(ab′)2 fragment. 
     Anti-CD71 (Fab)-PMO Conjugation 
     The F(ab′)2 fragment (15 mg/mL) in borate buffer (pH 8.0) was reduced by adding 10 equivalents of TCEP in water and incubating at 37° C. for 2 hours. SMCC was added to the primary amine on the 3′ end of the PMO by incubating the PMO (50 mg/mL) in DMSO with 10 equivalents of SMCC (10 mg/mL) in DMSO for 1 hour. Unconjugated SMCC was removed by ultrafiltration using Amicon Ultra-15 centrifugal filter units with a MWCO of 3 kDa. The PMO-SMCC was washed three times with acetate buffer (pH 6.0) and used immediately. The reduced F(ab′) fragment (Fab) was buffer exchanged into borate buffer (pH 8.0) using Amicon Ultra-15 Centrifugal Filter Units with a MWCO of 10 kDa, and 1.75 equivalents of PMO-SMCC was added and incubated overnight at 4° C. The pH of the reaction mixture was then reduced to 7.5, and 6 equivalents of N-Ethylmaleimide was added to the mixture at room temperature for 30 minutes to quench unreacted cysteines. Analysis of the reaction mixture by hydrophobic interaction chromatography (HIC) method 3 showed anti-CD71 (Fab)-PMO conjugates along with unreacted Fab ( FIG. 7A ).  FIG. 7A  shows a chromatogram of FPLC purification of anti-CD71 Fab-PMO using HIC method 3. 
     Purification 
     The reaction mixture was purified with an AKTA Explorer FPLC using HIC method 3. Fractions containing conjugates with a DAR of one, two and three were combined and concentrated separately. Concentrated conjugates were buffer exchanged with PBS (pH 7.4) using Amicon Ultra-15 centrifugal filter units with a MWCO of 10 kDa prior to analysis. 
     Analysis of the Purified Conjugate 
     The isolated conjugates were characterized by SEC, and HIC. SEC method 1 was used to confirm the absence of high molecular weight aggregates and unconjugated PMO. See  FIGS. 7B-7E .  FIG. 7B  shows a chromatogram of anti-CD71 Fab produced using SEC method 1.  FIG. 7C  shows a chromatogram of anti-CD71 Fab-PMO DAR 1 conjugate produced using SEC method 1.  FIG. 7D  shows a chromatogram of anti-CD71 Fab-PMO DAR 2 conjugate produced using SEC method 1.  FIG. 7E  shows a chromatogram of anti-CD71 Fab-PMO DAR 3 conjugate produced using SEC method 1. The purity of the conjugate was assessed by analytical HPLC using HIC method 4. See  FIGS. 7F-7I .  FIG. 7F  shows a chromatogram of anti-CD71 Fab produced using HIC method 4.  FIG. 7G  shows a chromatogram of anti-CD71 Fab-PMO DAR 1 conjugate produced using HIC method 4.  FIG. 7H  shows a chromatogram of anti-CD71 Fab-PMO DAR 2 conjugate produced using HIC method 4.  FIG. 7I  shows a chromatogram of anti-CD71 Fab-PMO DAR 3 conjugate produced using HIC method 4. “DAR” refers to drug-to-antibody ratio. The 260/280 nm UV absorbance ratio of each sample was compared to a standard curve of known ratios of PMO and Fab to confirm DAR. 
     Anti-CD71 Antibody Phosphorothioate Antisense Oligonucleotide Conjugate (Anti-CD71 mAb-PS ASO) 
     Anti-CD71 mAb-PSASO 
     Anti-CD71 antibody (10 mg/mL) in borate buffer (pH 8.0) was reduced by adding 4 equivalents of TCEP in water and incubating at 37° C. for 4 hours. 4(N-Maleimidomethyl)cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) was added to the primary amine on the 5′ end of the PS-ASO by incubating the PS ASO (50 mg/mL) in 1:1 mixture of 250 mM PB (pH 7.5) and DMSO with 10 equivalents of SMCC (10 mg/mL) in DMSO for 1 hour. Unconjugated SMCC was removed by ultrafiltration using Amicon Ultra-15 centrifugal filter units with a MWCO of 3 kDa. The PS ASO-SMCC was washed three times with acetate buffer (pH 6.0) and used immediately. The reduced antibody was mixed with 1.7 equivalents of PS ASO-SMCC and incubated overnight at 4° C. The pH of the reaction mixture was then reduced to 7.4, and 8 equivalents of N-Ethylmaleimide was added to the mixture at room temperature for 30 minutes to quench unreacted cysteines. Analysis of the reaction mixture by strong anion exchange chromatography (SAX) method 2 showed antibody-PS ASO conjugates along with unreacted antibody and ASO ( FIG. 8A ).  FIG. 8A  shows a chromatogram of anti-CD71 mAb-PS ASO reaction mixture produced with SAX method 2 showing free antibody peak (1), free PS ASO (5), DAR 1 (2), DAR 2 (3), DAR&gt;2 (4). “DAR” refers to a drug-to-antibody ratio. The number in parentheses refers to the peak. 
     Purification 
     The reaction mixture was purified with an AKTA Explorer FPLC using SAX method 1. Fractions containing conjugates with a drug-to-antibody ratio (DAR) of one, two and three were combined and concentrated separately and buffer exchanged with PBS (pH 7.4) using Amicon Ultra-15 centrifugal filter units with a MWCO of 50 kDa prior to analysis. 
     Analysis of the Purified Conjugate 
     The isolated conjugates were characterized by size exclusion chromatography (SEC) and SAX. Size exclusion chromatography method 1 was used to confirm the absence of high molecular weight aggregates and unconjugated ASO. See  FIGS. 8B-8E .  FIG. 8B  shows a chromatogram of anti-CD71 mAb produced using SEC method 1.  FIG. 8C  shows a chromatogram of anti-CD71 mAb-PS ASO DAR 1 conjugate produced using SEC method 1.  FIG. 8D  shows a chromatogram of anti-CD71 mAb-PS ASO DAR 2 conjugate produced using SEC method 1.  FIG. 8E  shows a chromatogram of anti-CD71 mAb-PS ASO DAR 3 conjugate produced using SEC method 1. The purity of the conjugate was assessed by analytical HPLC using SAX method 2. See  FIGS. 8F-8H .  FIG. 8F  shows a chromatogram of anti-CD71 mAb-PS ASO DAR 1 conjugate produced using SAX method 2.  FIG. 8G  shows a chromatogram of anti-CD71 mAb-PS ASO DAR 2 conjugate produced using SAX method 2.  FIG. 8H  shows a chromatogram of anti-CD71 mAb-PS ASO DAR 3 conjugate produced using SAX method 2. The 260/280 nm UV absorbance ratio of each sample was compared to a standard curve of known ratios of ASO and antibody to confirm drug-to-antibody ratio (DAR). 
     Example 4: In Vitro Activity of Anti-CD71 mAb-PMO Conjugate 
     The anti-CD71 mAb-PMO conjugate was made and characterized as described in Example 3. The conjugate was assessed for its ability to mediate exon skipping in vitro in differentiated C2C12 cells using nested PCR using methods similar to Example 2. Briefly, the potency of “naked” morpholino ASO (“PMO”) was compared to an anti-CD71 mAb-PMO conjugate at multiple concentrations with the relevant vehicle controls. Controls included vehicle (“Veh”), scramble morpholino at 50 uM (“Scr50”), and no antibody (“Neg-Ab”). The concentrations of PMO used included 50 uM, 1 uM, and 0.02 uM. The concentrations of anti-CD71 mAB-PMO DAR 1,2 used included 200 nM, 20 nM, and 2 nM. “DAR” refers to drug-to-antibody ratio. 
     Following cDNA synthesis, two rounds of PCR amplification (primary and nested PCR) were used to detect exon-skipping. PCR reactions were analyzed in a 4% TAE agarose gel ( FIG. 9 ). 
     Referring to  FIG. 9 , anti-CD71 mAb-PMO conjugate produced measurable exon 23 skipping in differentiated C2C12 cells and lower concentrations than the “naked” PMO control. The wild-type product had an expected size of 788 base pairs and the skipped DMD A23 of 575 base pairs. 
     A second experiment included an anti-CD71 Fab-PMO conjugate and a PMO targeted with an anti-EGFR (“Z-PMO”) as a negative control ( FIG. 10 ). The concentrations of PMO used included 10 uM and 2 uM. The concentrations of anti-CD71 mAb-PMO used included 0.2 uM and 0.04 uM. Anti-CD71 mAb-PMO had a DAR of 2. Z-PMO was used at a concentration of 0.2 uM and had a DAR of 2. Concentrations of anti-CD71 Fab-PMO included 0.6 uM and 0.12 uM. DAR of 1, 2, and 3 for anti-CD71 mAb-PMO at 0.6 uM and 0.12 uM were assayed. 
     Referring to  FIG. 10 , Receptor mediated uptake utilizing the transferrin receptor, the anti-CD71 mAb-PMO, and anti-CD71 Fab-PMO conjugates resulted in measurable exon 23 skipping in C2C12 cells and lower concentrations than the “naked” PMO control. There was no measurable exon 23 skipping from the Z-PMO at the concentration tested, which produced skipping from the anti-CD71 conjugates. 
     Example 5. In Vitro Activity of Anti-CD71-ASO mAb PS Conjugate 
     The anti-CD71 mAb-PS ASO conjugate was made and characterized as described in Example 3. The conjugate was assessed for its ability to mediate exon skipping in vitro in differentiated C2C12 cells using nested PCR using similar methods as described in Example 2. Briefly, the potency of “naked” phosphorothioate ASO (PS ASO) was compared to an anti-CD71 mAb-PS ASO conjugate at multiple concentrations, with the relevant vehicle control. Two rounds of of PCR amplification (primary and nested PCR) were performed following cDNA synthesis to detect exon-skipping. PCR reactions were analyzed in a 4% TAE agarose gel ( FIG. 11 ).  FIG. 11  shows an agarose gel of PMO, ASO, conjugated anti-CD71 mAb-ASO of DAR1 (“ASC-DAR1”), conjugated anti-CD71 mAb-ASO of DAR2 (“ASC-DAR2”), and conjugated anti-CD71 mAb-ASO of DAR3 (“ASC-DAR3”). “PMO” and “ASO” refers to free PMO and ASO, unconjugated to antibody. “Veh” refers to vehicle only. The concentrations tested included 0.2, 1, and 5 micromolar (M). 
     Referring to  FIG. 11 , the anti-CD71 mAb-PS ASO conjugate produced measurable exon 23 skipping in differentiated C2C12 cells and lower concentrations than the “naked” PS ASO control. The wild-type product had an expected size of 788 base pairs and the skipped DMD A23 of 575 base pairs. 
     Example 6: In Vivo Activity of Anti-CD71 mAb-PMO Conjugate 
     The anti-CD71 mAb-PMO conjugate was made and characterized as described in Example 3. The conjugate anti-CD71 mAb-PMO DAR1,2 anti-CD71 and mAb-PMO DAR&gt;2 were assessed for its ability to mediate exon skipping in vivo in wild-type CD-1 mice using similar methods as described in Example 2. “DAR” refers to drug-to-antibody ratio. 
     Mice were dosed via intravenous (iv) injection with the mAb, vehicle control, and antisense conjugates (ASCs) at the doses as provided in Table 12. “DAR” refers to drug-to-antibody ratio. The “naked” PMO was dosed via intramuscular injection into the gastrocnemius muscle at the doses provided in Table 12. After 4, 7, or 14 days, heart and gastrocnemius muscle tissues were harvested and snap-frozen in liquid nitrogen. RNAs were isolated, reversed transcribed and a nested PCR reactions were performed. PCR reactions were analyzed in 4% TAE agarose gels which were then quantitated by densitometry. 
     
       
         
           
               
             
               
                 TABLE 12 
               
             
            
               
                   
               
               
                 In vivo study design 
               
            
           
           
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 PMO:mAb 
                 Harvest 
               
               
                   
                   
                   
                 mAb dose 
                 PMO Dose 
                 Ratio 
                 Time 
               
               
                 Group 
                 Test Article 
                 N 
                 (mg/kg) 
                 (mg/kg) 
                 (mol/mol) 
                 (h) 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
            
               
                 1 
                 anti-CD71 mAb-PMO, DAR1, 2 
                 3 
                 50 
                 4.8 
                 1.6 
                 96 
               
               
                 2 
                 anti-CD71 mAb-PMO, DAR1, 2 
                 3 
                 50 
                 4.8 
                 1.6 
                 168 
               
               
                 3 
                 anti-CD71 mAb-PMO, DAR1, 2 
                 3 
                 50 
                 4.8 
                 1.6 
                 336 
               
               
                 4 
                 anti-CD71 mAb-PMO, DAR &gt; 2 
                 3 
                 50 
                 10.5 
                 3.7 
                 96 
               
               
                 5 
                 anti-CD71 mAb-PMO, DAR &gt; 2 
                 3 
                 50 
                 10.5 
                 3.7 
                 168 
               
               
                 6 
                 anti-CD71 mAb-PMO, DAR &gt; 2 
                 3 
                 50 
                 10.5 
                 3.7 
                 336 
               
               
                 7 
                 anti-CD71 mAb 
                 3 
                 50 
                   
                   
                 96 
               
               
                 8 
                 anti-CD71 mAb 
                 3 
                 50 
                   
                   
                 168 
               
               
                 9 
                 anti-CD71 mAb 
                 3 
                 50 
                   
                   
                 336 
               
               
                 10 
                 PMO 
                 3 
                 40 ug/inj. 
                   
                   
                 96 
               
               
                 11 
                 PMO 
                 3 
                 40 ug/inj. 
                   
                   
                 168 
               
               
                 12 
                 PMO 
                 3 
                 40 ug/inj. 
                   
                   
                 336 
               
               
                 13 
                 Vehicle 
                 3 
                   
                   
                   
                 96 
               
               
                 14 
                 Vehicle 
                 3 
                   
                   
                   
                 168 
               
               
                 15 
                 Vehicle 
                 3 
                   
                   
                   
                 336 
               
               
                   
               
            
           
         
       
     
       FIG. 12A  shows a gel electrophoresis of gastrocnemius muscle samples from mice administered anti-CD71 mAb-PMO DAR 1,2, anti-CD71 mAb-PMO DAR&gt;2, anti-CD71 mAb, PMO, and vehicle for 4, 7, or 14 days. The wild-type product had an expected size of 788 base pairs and the skipped DMD A23 of 575 base pairs. Anti-CD71 mAb-PMO DAR 1,2 and anti-CD71 mAb-PMO DAR&gt;2 produced measurable exon 23 skipping in gastrocnemius muscle and lower concentrations than the “naked” PMO control. The intensity of the bands on the gel ( FIG. 12A ) was quantitated by densitometry as seen in  FIG. 12B .  FIG. 12C  shows the quantification of in vivo exon skipping in wild-type mice gastrocnemius muscle using Taqman qPCR. 
       FIG. 13A  shows a gel electrophoresis of heart samples from mice administered anti-CD71 mAb-PMO DAR 1,2, anti-CD71 mAb-PMO DAR&gt;2, anti-CD71 mAb, PMO, and vehicle for 4, 7, or 14 days. The wild-type product had an expected size of 788 base pairs and the skipped DMD A23 of 575 base pairs. The intensity of the bands on the gel ( FIG. 13A ) was quantitated by densitometry as seen in  FIG. 13B . Similar results as with the gastrocnemius muscle samples were obtained. Anti-CD71 mAb-PMO DAR 1,2 and anti-CD71 mAb-PMO DAR&gt;2 produced measurable exon 23 skipping in gastrocnemius muscle and lower concentrations than the “naked” PMO control. 
     DNA fragments were then isolated from the 4% agarose gels and sequenced. The sequencing data confirmed the correct sequence in the skipped and wild-type products as seen in  FIG. 14 . 
     Example 7. Sequences 
     Table 13 illustrates exemplary target sequences to induce insertion, deletion, duplications, or alteration in the DMD gene using compositions and methods as described herein. Table 14 illustrates exemplary nucleotide sequences to induce an insertion, deletion, duplication, or alteration in the DMD gene using compositions and methods as described herein. Table 15 and Table 16 illustrate exemplary target sequences in several genes for inducing an insertion, deletion, duplications, or alteration in the gene. Table 17 illustrates exemplary sequences, including sequences in the DMD gene to induce an insertion, deletion, duplication, or alteration in the gene using compositions and methods as described herein. 
     
       
         
           
               
               
               
             
               
                 TABLE 13 
               
               
                   
               
               
                   
                   
                 SEQ 
               
               
                 Target 
                   
                 ID 
               
               
                 Exon 
                 Antisense Sequence 
                 NO. 
               
               
                   
               
             
            
               
                 19 
                 5′ GCCUGAGCUGAUCUGCUGGCAUCUUGCAGU 
                 45 
               
               
                   
                 U 3′ 
                   
               
               
                   
               
               
                 19 or 20 
                 5′GCAGAAUUCGAUCCACCGGCUGUUCAAGCCU 
                 46 
               
               
                   
                 GAGCUGAUCUGCUCGCAUCUUGCAGU3′ 
                   
               
               
                   
               
               
                 20 
                 5′ CAGCAGUAGUUGUCAUCUGCUC 3′ 
                 47 
               
               
                   
               
               
                 21 
                 5′ CACAAAGUCUGCAUCCAGGAACAUGGGUC  
                 48 
               
               
                   
                 3′ 
                   
               
               
                   
               
               
                 22 
                 5′ CUGCAAUUCCCCGAGUCUCUGC 3′ 
                 49 
               
               
                   
               
               
                 51 
                 5′ CUCAUACCUUCUGCUUGAUGAUC 3′ 
                 50 
               
               
                   
               
               
                 52 
                 5′ UCCAACUGGGGACGCCUCUGUUCCAAAUCC 
                 51 
               
               
                   
                 3′ 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
             
               
                 TABLE 14 
               
               
                   
               
               
                   
                   
                   
                 SEQ 
               
               
                   
                   
                   
                 ID 
               
               
                 Gene 
                 Target Location 
                 Nucleotide Sequence (5′-3′) 
                 NO. 
               
               
                   
               
             
            
               
                   
               
            
           
           
               
               
               
               
            
               
                 DMD 
                 H8A(−06+18) 
                 GAUAGGUGGUAUCAACAUCUGUAA 
                 52 
               
               
                   
               
               
                 DMD 
                 H8A(−03+18) 
                 GAUAGGUGGUAUCAACAUCUG 
                 53 
               
               
                   
               
               
                 DMD 
                 H8A(−07+18) 
                 GAUAGGUGGUAUCAACAUCUGUAAG 
                 54 
               
               
                   
               
               
                 DMD 
                 H8A(−06+14) 
                 GGUGGUAUCAACAUCUGUAA 
                 55 
               
               
                   
               
               
                 DMD 
                 H8A(−10+10) 
                 GUAUCAACAUCUGUAAGCAC 
                 56 
               
               
                   
               
               
                 DMD 
                 H7A(+45+67) 
                 UGCAUGUUCCAGUCGUUGUGUGG 
                 57 
               
               
                   
               
               
                 DMD 
                 H7A(+02+26) 
                 CACUAUUCCAGUCAAAUAGGUCUGG 
                 58 
               
               
                   
               
               
                 DMD 
                 H7D(+15−10) 
                 AUUUACCAACCUUCAGGAUCGAGUA 
                 59 
               
               
                   
               
               
                 DMD 
                 H7A(−18+03) 
                 GGCCUAAAACACAUACACAUA 
                 60 
               
               
                   
               
               
                 DMD 
                 C6A(−10+10) 
                 CAUUUUUGACCUACAUGUGG 
                 61 
               
               
                   
               
               
                 DMD 
                 C6A(−14+06) 
                 UUUGACCUACAUGUGGAAAG 
                 62 
               
               
                   
               
               
                 DMD 
                 C6A(−14+12) 
                 UACAUUUUUGACCUACAUGUGGAAAG 
                 63 
               
               
                   
               
               
                 DMD 
                 C6A(−13+09) 
                 AUUUUUGACCUACAUGGGAAAG 
                 64 
               
               
                   
               
               
                 DMD 
                 CH6A(+69+91) 
                 UACGAGUUGAUUGUCGGACCCAG 
                 65 
               
               
                   
               
               
                 DMD 
                 C6D(+12−13) 
                 GUGGUCUCCUUACCUAUGACUGUGG 
                 66 
               
               
                   
               
               
                 DMD 
                 C6D(+06−11) 
                 GGUCUCCUUACCUAUGA 
                 67 
               
               
                   
               
               
                 DMD 
                 H6D(+04−21) 
                 UGUCUCAGUAAUCUUCUUACCUAU 
                 68 
               
               
                   
               
               
                 DMD 
                 H6D(+18−04) 
                 UCUUACCUAUGACUAUGGAUGAGA 
                 69 
               
               
                   
               
               
                 DMD 
                 H4A(+13+32) 
                 GCAUGAACUCUUGUGGAUCC 
                 70 
               
               
                   
               
               
                 DMD 
                 H4D(+04−16) 
                 CCAGGGUACUACUUACAUUA 
                 71 
               
               
                   
               
               
                 DMD 
                 H4D(−24−44) 
                 AUCGUGUGUCACAGCAUCCAG 
                 72 
               
               
                   
               
               
                 DMD 
                 H4A(+11+40) 
                 UGUUCAGGGCAUGAACUCUUGUGGAUCCUU 
                 73 
               
               
                   
               
               
                 DMD 
                 H3A(+30+60) 
                 UAGGAGGCGCCUCCCAUCCUGUAGGUCACUG 
                 74 
               
               
                   
               
               
                 DMD 
                 H3A(+35+65) 
                 AGGUCUAGGAGGCGCCUCCCAUCCUGUAGGU 
                 75 
               
               
                   
               
               
                 DMD 
                 H3A(+30+54) 
                 GCGCCUCCCAUCCUGUAGGUCACUG 
                 76 
               
               
                   
               
               
                 DMD 
                 H3D(+46−21) 
                 CUUCGAGGAGGUCUAGGAGGCGCCUC 
                 77 
               
               
                   
               
               
                 DMD 
                 H3A(+30+50) 
                 CUCCCAUCCUGUAGGUCACUG 
                 78 
               
               
                   
               
               
                 DMD 
                 H3D(+19−03) 
                 UACCAGUUUUUGCCCUGUCAGG 
                 79 
               
               
                   
               
               
                 DMD 
                 H3A(−06+20) 
                 UCAAUAUGCUGCUUCCCAAACUGAAA 
                 80 
               
               
                   
               
               
                 DMD 
                 H3A(+37+61) 
                 CUAGGAGGCGCCUCCCAUCCUGUAG 
                 81 
               
               
                   
               
               
                 DMD 
                 H5A(+20+50) 
                 UUAUGAUUUCCAUCUACGAUGUCAGUACUUC 
                 82 
               
               
                   
               
               
                 DMD 
                 H5D(+25−05) 
                 CUUACCUGCCAGUGGAGGAUUAUAUUCCAAA 
                 83 
               
               
                   
               
               
                 DMD 
                 H5D(+10−15) 
                 CAUCAGGAUUCUUACCUGCCAGUGG 
                 84 
               
               
                   
               
               
                 DMD 
                 H5A(+10+34) 
                 CGAUGUCAGUACUUCCAAUAUUCAC 
                 85 
               
               
                   
               
               
                 DMD 
                 H5D(−04−21) 
                 ACCAUUCAUCAGGAUUCU 
                 86 
               
               
                   
               
               
                 DMD 
                 H5D(+16−02) 
                 ACCUGCCAGUGGAGGAUU 
                 87 
               
               
                   
               
               
                 DMD 
                 H5A(−07+20) 
                 CCAAUAUUCACUAAAUCAACCUGUUAA 
                 88 
               
               
                   
               
               
                 DMD 
                 H5D(+18−12) 
                 CAGGAUUGUUACCUGCCAGUGGAGGAUUAU 
                 89 
               
               
                   
               
               
                 DMD 
                 H5A(+05+35) 
                 ACGAUGUCAGUACUUCCAAUAUUCACUAAAU 
                 90 
               
               
                   
               
               
                 DMD 
                 H5A(+15+45) 
                 AUUUCCAUCUACGAUGUCAGUACUUCCAAUA 
                 91 
               
               
                   
               
               
                 DMD 
                 H10A(−05+16) 
                 CAGGAGCUUCCAAAUGCUGCA 
                 92 
               
               
                   
               
               
                 DMD 
                 H10A(−05+24) 
                 CUUGUCUUCAGGAGCUUCCAAAUGCUGCA 
                 93 
               
               
                   
               
               
                 DMD 
                 H10A(+98+119) 
                 UCCUCAGCAGAAAGAAGCCACG 
                 94 
               
               
                   
               
               
                 DMD 
                 H10A(+130+149) 
                 UUAGAAAUCUCUCCUUGUGC 
                 95 
               
               
                   
               
               
                 DMD 
                 H10A(−33−14) 
                 UAAAUUGGGUGUUACACAAU 
                 96 
               
               
                   
               
               
                 DMD 
                 H11D(+26+49) 
                 CCCUGAGGCAUUCCCAUCUUGAAU 
                 97 
               
               
                   
               
               
                 DMD 
                 H11D(+11−09) 
                 AGGACUUACUUGCUUUGUUU 
                 98 
               
               
                   
               
               
                 DMD 
                 H11A(+118+140) 
                 CUUGAAUUUAGGAGAUUCAUCUG 
                 99 
               
               
                   
               
               
                 DMD 
                 H11A(+75+97) 
                 CAUCUUCUGAUAAUUUUCCUGUU 
                 100 
               
               
                   
               
               
                 DMD 
                 H12A(+52+75) 
                 UCUUCUGUUUUUGUUAGCCAGUCA 
                 101 
               
               
                   
               
               
                 DMD 
                 H12A(−10+10) 
                 UCUAUGUAAACUGAAAAUUU 
                 102 
               
               
                   
               
               
                 DMD 
                 H12A(+11+30) 
                 UUCUGGAGAUCCAUUAAAAC 
                 103 
               
               
                   
               
               
                 DMD 
                 H13A(+77+100) 
                 CAGCAGUUGCGUGAUCUCCACUAG 
                 104 
               
               
                   
               
               
                 DMD 
                 H13A(+55+75) 
                 UUCAUCAACUACCACCACCAU 
                 105 
               
               
                   
               
               
                 DMD 
                 H13D(+06−19) 
                 CUAAGCAAAAUAAUCUGACCUUAAG 
                 106 
               
               
                   
               
               
                 DMD 
                 H14A(+37+64) 
                 CUUGUAAAAGAACCCAGCGGUCUUCUGU 
                 107 
               
               
                   
               
               
                 DMD 
                 H14A(+14+35) 
                 CAUCUACAGAUGUUUGCCCAUC 
                 108 
               
               
                   
               
               
                 DMD 
                 H14A(+51+73) 
                 GAAGGAUGUCUUGUAAAAGAACC 
                 109 
               
               
                   
               
               
                 DMD 
                 H14D(−02+18) 
                 ACCUGUUCUUCAGUAAGACG 
                 110 
               
               
                   
               
               
                 DMD 
                 H14D(+14−10) 
                 CAUGACACACCUGUUCUUCAGUAA 
                 111 
               
               
                   
               
               
                 DMD 
                 H14A(+61+80) 
                 CAUUUGAGAAGGAUGUCUUG 
                 112 
               
               
                   
               
               
                 DMD 
                 H14A(−12+12) 
                 AUCUCCCAAUACCUGGAGAAGAGA 
                 113 
               
               
                   
               
               
                 DMD 
                 H15A(−12+19) 
                 GCCAUGCACUAAAAAGGCACUGCAAGACAUU 
                 114 
               
               
                   
               
               
                 DMD 
                 H15A(+48+71) 
                 UCUUUAAAGCCAGUUGUGUGAAUC 
                 115 
               
               
                   
               
               
                 DMD 
                 H15A(+08+28) 
                 UUUCUGAAAGCCAUGCACUAA 
                 116 
               
               
                   
               
               
                 DMD 
                 H15D(+17−08) 
                 GUACAUACGGCCAGUUUUUGAAGAC 
                 117 
               
               
                   
               
               
                 DMD 
                 H16A(−12+19) 
                 CUAGAUCCGCUUUUAAAACCUGUUAAAACAA 
                 118 
               
               
                   
               
               
                 DMD 
                 H16A(−06+25) 
                 UCUUUUCUAGAUCCGCUUUUAAAACCUGUUA 
                 119 
               
               
                   
               
               
                 DMD 
                 H16A(−06+19) 
                 CUAGAUCCGCUUUUAAAACCUGUUA 
                 120 
               
               
                   
               
               
                 DMD 
                 H16A(+87+109) 
                 CCGUCUUCUGGGUCACUGACUUA 
                 121 
               
               
                   
               
               
                 DMD 
                 H16A(−07+19) 
                 CUAGAUCCGCUUUUAAAACCUGUUAA 
                 122 
               
               
                   
               
               
                 DMD 
                 H16A(−07+13) 
                 CCGCUUUUAAAACCUGUUAA 
                 123 
               
               
                   
               
               
                 DMD 
                 H16A(+12+37) 
                 UGGAUUGCUUUUUCUUUUCUAGAUCC 
                 124 
               
               
                   
               
               
                 DMD 
                 H16A(+92+116) 
                 CAUGCUUCCGUCUUCUGGGUCACUG 
                 125 
               
               
                   
               
               
                 DMD 
                 H16A(+45+67) 
                 GAUCUUGUUUGAGUGAAUACAGU 
                 126 
               
               
                   
               
               
                 DMD 
                 H16A(+105+126) 
                 GUUAUCCAGCCAUGCUUCCGUC 
                 127 
               
               
                   
               
               
                 DMD 
                 H16D(+05−20) 
                 UGAUAAUUGGUAUCACUAACCUGUG 
                 128 
               
               
                   
               
               
                 DMD 
                 H16D(+12−11) 
                 GUAUCACUAACCUGUGCUGUAC 
                 129 
               
               
                   
               
               
                 DMD 
                 H19A(+35+53) 
                 CUGCUGGCAUCUUGCAGUU 
                 130 
               
               
                   
               
               
                 DMD 
                 H19A(+35+65) 
                 GCCUGAGCUGAUCUGCUGGCAUCUUGCAGUU 
                 131 
               
               
                   
               
               
                 DMD 
                 H20A(+44+71) 
                 CUGGCAGAAUUCGAUCCACCGGCUGUUC 
                 132 
               
               
                   
               
               
                 DMD 
                 H20A(+147+168) 
                 CAGCAGUAGUUGUCAUCUGCUC 
                 133 
               
               
                   
               
               
                 DMD 
                 H20A(+185+203) 
                 UGAUGGGGUGGUGGGUUGG 
                 134 
               
               
                   
               
               
                 DMD 
                 H20A(−08+17) 
                 AUCUGCAUUAACACCCUCUAGAAAG 
                 135 
               
               
                   
               
               
                 DMD 
                 H20A(+30+53) 
                 CCGGCUGUUCAGUUGUUCUGAGGC 
                 136 
               
               
                   
               
               
                 DMD 
                 H20A(−11+17) 
                 AUCUGCAUUAACACCCUCUAGAAAGAAA 
                 137 
               
               
                   
               
               
                 DMD 
                 H20D(+08−20) 
                 GAAGGAGAAGAGAUUCUUACCUUACAAA 
                 138 
               
               
                   
               
               
                 DMD 
                 H20A(+44+63) 
                 AUUCGAUCCACCGGCUGUUC 
                 139 
               
               
                   
               
               
                 DMD 
                 H20A(+149+168 
                 CAGCAGUAGUUGUCAUCUGC 
                 140 
               
               
                   
               
               
                 DMD 
                 H21A(−06+16) 
                 GCCGGUUGACUUCAUCCUGUGC 
                 141 
               
               
                   
               
               
                 DMD 
                 H21A(+85+106) 
                 CUGCAUCCAGGAACAUGGGUCC 
                 142 
               
               
                   
               
               
                 DMD 
                 H21A(+85+108) 
                 GUCUGCAUCCAGGAACAUGGGUC 
                 143 
               
               
                   
               
               
                 DMD 
                 H21A(+08+31) 
                 GUUGAAGAUCUGAUAGCCGGUUGA 
                 144 
               
               
                   
               
               
                 DMD 
                 H21D(+18−07) 
                 UACUUACUGUCUGUAGCUCUUUCU 
                 145 
               
               
                   
               
               
                 DMD 
                 H22A(+22+45) 
                 CACUCAUGGUCUCCUGAUAGCGCA 
                 146 
               
               
                   
               
               
                 DMD 
                 H22A(+125+106) 
                 CUGCAAUUCCCCGAGUCUCUGC 
                 147 
               
               
                   
               
               
                 DMD 
                 H22A(+47+69) 
                 ACUGCUGGACCCAUGUCCUGAUG 
                 148 
               
               
                   
               
               
                 DMD 
                 H22A(+80+101) 
                 CUAAGUUGAGGUAUGGAGAGU 
                 149 
               
               
                   
               
               
                 DMD 
                 H22D(+13−11) 
                 UAUUCACAGACCUGCAAUUCCCC 
                 150 
               
               
                   
               
               
                 DMD 
                 H23A(+34+59) 
                 ACAGUGGUGCUGAGAUAGUAUAGGCC 
                 151 
               
               
                   
               
               
                 DMD 
                 H23A(+18+39) 
                 UAGGCCACUUUGUUGCUCUUGC 
                 152 
               
               
                   
               
               
                 DMD 
                 H23A(+72+90) 
                 UUCAGAGGGCGCUUUCUUC 
                 153 
               
               
                   
               
               
                 DMD 
                 H24A(+48+70) 
                 GGGCAGGCCAUUCCUCCUUCAGA 
                 154 
               
               
                   
               
               
                 DMD 
                 H24A(−02+22) 
                 UCUUCAGGGUUUGUAUGUGAUUCU 
                 155 
               
               
                   
               
               
                 DMD 
                 H25A(+9+36) 
                 CUGGGCUGAAUUGUCUGAAUAUCACUG 
                 156 
               
               
                   
               
               
                 DMD 
                 H25A(+131+156) 
                 CUGUUGGCACAUGUGAUCCCACUGAG 
                 157 
               
               
                   
               
               
                 DMD 
                 H25D(+16−08) 
                 GUCUAUACCUGUUGGCACAUGUGA 
                 158 
               
               
                   
               
               
                 DMD 
                 H26A(+132+156) 
                 UGCUUUCUGUAAUUCAUCUGGAGUU 
                 159 
               
               
                   
               
               
                 DMD 
                 H26A(−07+19) 
                 CCUCCUUUCUGGCAUAGACCUUCCAC 
                 160 
               
               
                   
               
               
                 DMD 
                 H26A(+68+92) 
                 UGUGUCAUCCAUUCGUGCAUCUCUG 
                 161 
               
               
                   
               
               
                 DMD 
                 H27A(+82+106) 
                 UUAAGGCCUCUUGUGCUACAGGUGG 
                 162 
               
               
                   
               
               
                 DMD 
                 H27A(−4+19) 
                 GGGGCUCUUCUUUAGCUCUCUGA 
                 163 
               
               
                   
               
               
                 DMD 
                 H27D(+19−03) 
                 GACUUCCAAAGUCUUGCAUUUC 
                 164 
               
               
                   
               
               
                 DMD 
                 H28A(−05+19) 
                 GCCAACAUGCCCAAACUUCCUAAG 
                 165 
               
               
                   
               
               
                 DMD 
                 H28A(+99+124) 
                 CAGAGAUUUCCUCAGCUCCGCCAGGA 
                 166 
               
               
                   
               
               
                 DMD 
                 H28D(+16−05) 
                 CUUACAUCUAGCACCUCAGAG 
                 167 
               
               
                   
               
               
                 DMD 
                 H29A(+57+81) 
                 UCCGCCAUCUGUUAGGGUCUGUGCC 
                 168 
               
               
                   
               
               
                 DMD 
                 H29A(+18+42) 
                 AUUUGGGUUAUCCUCUGAAUGUCGC 
                 169 
               
               
                   
               
               
                 DMD 
                 H29D(+17−05) 
                 CAUACCUCUUCAUGUAGUUCCC 
                 170 
               
               
                   
               
               
                 DMD 
                 H30A(+122+147) 
                 CAUUUGAGCUGCGUCCACCUUGUCUG 
                 171 
               
               
                   
               
               
                 DMD 
                 H30A(+25+50) 
                 UCCUGGGCAGACUGGAUGCUCUGUUC 
                 172 
               
               
                   
               
               
                 DMD 
                 H30D(+19−04) 
                 UUGCCUGGGCUUCCUGAGGCAUU 
                 173 
               
               
                   
               
               
                 DMD 
                 H31D(+06−18) 
                 UUCUGAAAUAACAUAUACCUGUGC 
                 174 
               
               
                   
               
               
                 DMD 
                 H31D(+03−22) 
                 UAGUUUCUGAAAUAACAUAUACCUG 
                 175 
               
               
                   
               
               
                 DMD 
                 H31A(+05+25) 
                 GACUUGUCAAAUCAGAUUGGA 
                 176 
               
               
                   
               
               
                 DMD 
                 H31D(+04−20) 
                 GUUUCUGAAAUAACAUAUACCUGU 
                 177 
               
               
                   
               
               
                 DMD 
                 H32D(+04−16) 
                 CACCAGAAAUACAUACCACA 
                 178 
               
               
                   
               
               
                 DMD 
                 H32A(+151+170) 
                 CAAUGAUUUAGCUGUGACUG 
                 179 
               
               
                   
               
               
                 DMD 
                 H32A(+10+32) 
                 CGAAACUUCAUGGAGACAUCUUG 
                 180 
               
               
                   
               
               
                 DMD 
                 H32A(+49+73) 
                 CUUGUAGACGCUGCUCAAAAUUGGC 
                 181 
               
               
                   
               
               
                 DMD 
                 H33D(+09−11) 
                 CAUGCACACACCUUUGCUCC 
                 182 
               
               
                   
               
               
                 DMD 
                 H33A(+53+76) 
                 UCUGUACAAUCUGACGUCCAGUCU 
                 183 
               
               
                   
               
               
                 DMD 
                 H33A(+30+56) 
                 GUCUUUAUCACCAUUUCCACUUCAGAC 
                 184 
               
               
                   
               
               
                 DMD 
                 H33A(+64+88) 
                 CCGUCUGCUUUUUCUGUACAAUCUG 
                 185 
               
               
                   
               
               
                 DMD 
                 H34A(+83+104) 
                 UCCAUAUCUGUAGCUGCCAGCC 
                 186 
               
               
                   
               
               
                 DMD 
                 H34A(+143+165) 
                 CCAGGCAACUUCAGAAUCCAAAU 
                 187 
               
               
                   
               
               
                 DMD 
                 H34A(−20+10) 
                 UUUCUGUUACCUGAAAAGAAUUAUAAUGAA 
                 188 
               
               
                   
               
               
                 DMD 
                 H34A(+46+70) 
                 CAUUCAUUUCCUUUCGCAUCUUACG 
                 189 
               
               
                   
               
               
                 DMD 
                 H34A(+95+120) 
                 UGAUCUCUUUGUCAAUUCCAUAUCUG 
                 190 
               
               
                   
               
               
                 DMD 
                 H34D(+10−20) 
                 UUCAGUGAUAUAGGUUUUACCUUUCCCCAG 
                 191 
               
               
                   
               
               
                 DMD 
                 H34A(+72+96) 
                 CUG UAG CUG CCA GCC AUU CUG UCA AG 
                 192 
               
               
                   
               
               
                 DMD 
                 H35A(+141+161) 
                 UCU UCU GCU CGG GAG GUG ACA 
                 193 
               
               
                   
               
               
                 DMD 
                 H35A(+116+135) 
                 CCA GUU ACU AUU CAG AAG AC 
                 194 
               
               
                   
               
               
                 DMD 
                 H35A(+24+43) 
                 UCU UCA GGU GCA CCU UCU GU 
                 195 
               
               
                   
               
               
                 DMD 
                 H36A(+26+50) 
                 UGUGAUGUGGUCCACAUUCUGGUCA 
                 196 
               
               
                   
               
               
                 DMD 
                 H36A(−02+18) 
                 CCAUGUGUUUCUGGUAUUCC 
                 197 
               
               
                   
               
               
                 DMD 
                 H37A(+26+50) 
                 CGUGUAGAGUCCACCUUUGGGCGUA 
                 198 
               
               
                   
               
               
                 DMD 
                 H37A(+82+105) 
                 UACUAAUUUCCUGCAGUGGUCACC 
                 199 
               
               
                   
               
               
                 DMD 
                 H37A(+134+157) 
                 UUCUGUGUGAAAUGGCUGCAAAUC 
                 200 
               
               
                   
               
               
                 DMD 
                 H38A(−01+19) 
                 CCUUCAAAGGAAUGGAGGCC 
                 201 
               
               
                   
               
               
                 DMD 
                 H38A(+59+83) 
                 UGCUGAAUUUCAGCCUCCAGUGGUU 
                 202 
               
               
                   
               
               
                 DMD 
                 H38A(+88+112) 
                 UGAAGUCUUCCUCUUUCAGAUUCAC 
                 203 
               
               
                   
               
               
                 DMD 
                 H39A(+62+85) 
                 CUGGCUUUCUCUCAUCUGUGAUUC 
                 204 
               
               
                   
               
               
                 DMD 
                 H39A(+39+58) 
                 GUUGUAAGUUGUCUCCUCUU 
                 205 
               
               
                   
               
               
                 DMD 
                 H39A(+102+121) 
                 UUGUCUGUAACAGCUGCUGU 
                 206 
               
               
                   
               
               
                 DMD 
                 H39D(+10−10) 
                 GCUCUAAUACCUUGAGAGCA 
                 207 
               
               
                   
               
               
                 DMD 
                 H40A(−05+17) 
                 CUUUGAGACCUCAAAUCCUGUU 
                 208 
               
               
                   
               
               
                 DMD 
                 H40A(+129+153) 
                 CUUUAUUUUCCUUUCAUCUCUGGGC 
                 209 
               
               
                   
               
               
                 DMD 
                 H42A(−04+23) 
                 AUCGUUUCUUCACGGACAGUGUGCUGG 
                 210 
               
               
                   
               
               
                 DMD 
                 H42A(+86+109) 
                 GGGCUUGUGAGACAUGAGUGAUUU 
                 211 
               
               
                   
               
               
                 DMD 
                 H42D(+19−02) 
                 ACCUUCAGAGGACUCCUCUUGC 
                 212 
               
               
                   
               
               
                 DMD 
                 H43D(+10−15) 
                 UAUGUGUUACCUACCCUUGUCGGUC 
                 213 
               
               
                   
               
               
                 DMD 
                 H43A(+101+120) 
                 GGAGAGAGCUUCCUGUAGCU 
                 214 
               
               
                   
               
               
                 DMD 
                 H43A(+78+100) 
                 UCACCCUUUCCACAGGCGUUGCA 
                 215 
               
               
                   
               
               
                 DMD 
                 H44A(+85+104) 
                 UUUGUGUCUUUCUGAGAAAC 
                 216 
               
               
                   
               
               
                 DMD 
                 H44D(+10−10) 
                 AAAGACUUACCUUAAGAUAC 
                 217 
               
               
                   
               
               
                 DMD 
                 H44A(−06+14) 
                 AUCUGUCAAAUCGCCUGCAG 
                 218 
               
               
                   
               
               
                 DMD 
                 H46D(+16−04) 
                 UUACCUUGACUUGCUCAAGC 
                 219 
               
               
                   
               
               
                 DMD 
                 H46A(+90+109) 
                 UCCAGGUUCAAGUGGGAUAC 
                 220 
               
               
                   
               
               
                 DMD 
                 H47A(+76+100) 
                 GCUCUUCUGGGCUUAUGGGAGCACU 
                 221 
               
               
                   
               
               
                 DMD 
                 H47D(+25−02) 
                 ACCUUUAUCCACUGGAGAUUUGUCUGC 
                 222 
               
               
                   
               
               
                 DMD 
                 H47A(−9+12) 
                 UUCCACCAGUAACUGAAACAG 
                 223 
               
               
                   
               
               
                 DMD 
                 H50A(+02+30) 
                 CCACUCAGAGCUCAGAUCUUCUAACUUCC 
                 224 
               
               
                   
               
               
                 DMD 
                 H50A(+07+33) 
                 CUUCCACUCAGAGCUCAGAUCUUCUAA 
                 225 
               
               
                   
               
               
                 DMD 
                 H50D(+07−18) 
                 GGGAUCCAGUAUACUUACAGGCUCC 
                 226 
               
               
                   
               
               
                 DMD 
                 H51A(−01+25) 
                 ACCAGAGUAACAGUCUGAGUAGGAGC 
                 227 
               
               
                   
               
               
                 DMD 
                 H51D(+16−07) 
                 CUCAUACCUUCUGCUUGAUGAUC 
                 228 
               
               
                   
               
               
                 DMD 
                 H51A(+111+134) 
                 UUCUGUCCAAGCCCGGUUGAAAUC 
                 229 
               
               
                   
               
               
                 DMD 
                 H51A(+61+90) 
                 ACAUCAAGGAAGAUGGCAUUUCUAGUUUGG 
                 230 
               
               
                   
               
               
                 DMD 
                 H51A(+66+90) 
                 ACAUCAAGGAAGAUGGCAUUUCUAG 
                 231 
               
               
                   
               
               
                 DMD 
                 H51A(+66+95) 
                 CUCCAACAUCAAGGAAGAUGGCAUUUCUAG 
                 232 
               
               
                   
               
               
                 DMD 
                 H51D(+08−17) 
                 AUCAUUUUUUCUCAUACCUUCUGCU 
                 233 
               
               
                   
               
               
                 DMD 
                 H51A/D(+08−17) 
                 AUCAUUUUUUCUCAUACCUUCUGCUAG 
                 234 
               
               
                   
               
               
                 DMD 
                 &amp;(−15+) 
                 GAGCUAAAA 
                 235 
               
               
                   
               
               
                 DMD 
                 H51A(+175+195) 
                 CACCCACCAUCACCCUCUGUG 
                 236 
               
               
                   
               
               
                 DMD 
                 H51A(+199+220) 
                 AUCAUCUCGUUGAUAUCCUCAA 
                 237 
               
               
                   
               
               
                 DMD 
                 H52A(−07+14) 
                 UCCUGCAUUGUUGCCUGUAAG 
                 238 
               
               
                   
               
               
                 DMD 
                 H52A(+12+41) 
                 UCCAACUGGGGACGCCUCUGUUCCAAAUCC 
                 239 
               
               
                   
               
               
                 DMD 
                 H52A(+17+37) 
                 ACUGGGGACGCCUCUGUUCCA 
                 240 
               
               
                   
               
               
                 DMD 
                 H52A(+93+112) 
                 CCGUAAUGAUUGUUCUAGCC 
                 241 
               
               
                   
               
               
                 DMD 
                 H52D(+05−15) 
                 UGUUAAAAAACUUACUUCGA 
                 242 
               
               
                   
               
               
                 DMD 
                 H53A(+45+69) 
                 CAUUCAACUGUUGCCUCCGGUUCUG 
                 243 
               
               
                   
               
               
                 DMD 
                 H53A(+39+62) 
                 CUGUUGCCUCCGGUUCUGAAGGUG 
                 244 
               
               
                   
               
               
                 DMD 
                 H53A(+39+69) 
                 CAUUCAACUGUUGCCUCCGGUUCUGAAGGUG 
                 245 
               
               
                   
               
               
                 DMD 
                 H53D(+14−07) 
                 UACUAACCUUGGUUUCUGUGA 
                 246 
               
               
                   
               
               
                 DMD 
                 H53A(+23+47) 
                 CUGAAGGUGUUCUUGUACUUCAUCC 
                 247 
               
               
                   
               
               
                 DMD 
                 H53A(+150+176) 
                 UGUAUAGGGACCCUCCUUCCAUGACUC 
                 248 
               
               
                   
               
               
                 DMD 
                 H53D(+20−05) 
                 CUAACCUUGGUUUCUGUGAUUUUCU 
                 249 
               
               
                   
               
               
                 DMD 
                 H53D(+09−18) 
                 GGUAUCUUUGAUACUAACCUUGGUUUC 
                 250 
               
               
                   
               
               
                 DMD 
                 H53A(−12+10) 
                 AUUCUUUCAACUAGAAUAAAAG 
                 251 
               
               
                   
               
               
                 DMD 
                 H53A(−07+18) 
                 GAUUCUGAAUUCUUUCAACUAGAAU 
                 252 
               
               
                   
               
               
                 DMD 
                 H53A(+07+26) 
                 AUCCCACUGAUUCUGAAUUC 
                 253 
               
               
                   
               
               
                 DMD 
                 H53A(+124+145) 
                 UUGGCUCUGGCCUGUCCUAAGA 
                 254 
               
               
                   
               
               
                 DMD 
                 H46A(+86+115) 
                 CUCUUUUCCAGGUUCAAGUGGGAUACUAGC 
                 255 
               
               
                   
               
               
                 DMD 
                 H46A(+107+137) 
                 CAAGCUUUUCUUUUAGUUGCUGCUCUUUUCC 
                 256 
               
               
                   
               
               
                 DMD 
                 H46A(−10+20) 
                 UAUUCUUUUGUUCUUCUAGCCUGGAGAAAG 
                 257 
               
               
                   
               
               
                 DMD 
                 H46A(+50+77) 
                 CUGCUUCCUCCAACCAUAAAACAAAUUC 
                 258 
               
               
                   
               
               
                 DMD 
                 H45A(−06+20) 
                 CCAAUGCCAUCCUGGAGUUCCUGUAA 
                 259 
               
               
                   
               
               
                 DMD 
                 H45A(+91+110) 
                 UCCUGUAGAAUACUGGCAUC 
                 260 
               
               
                   
               
               
                 DMD 
                 H45A(+125+151) 
                 UGCAGACCUCCUGCCACCGCAGAUUCA 
                 261 
               
               
                   
               
               
                 DMD 
                 H45D(+16−04) 
                 CUACCUCUUUUUUCUGUCUG 
                 262 
               
               
                   
               
               
                 DMD 
                 H45A(+71+90) 
                 UGUUUUUGAGGAUUGCUGAA 
                 263 
               
               
                   
               
               
                 * The first letter designates the species (e.g. H: human, M: murine, C: canine). 
               
               
                 “#” designates target DMD exon number. 
               
               
                 “A/D” indicates acceptor or donor splice site at the beginning and end of the exon, respectively. 
               
               
                 (x y) represents the annealing coordinates where “—“ or “+” indicate intronic or exonic sequences respectively. 
               
            
           
         
       
     
     
       
         
           
               
               
               
             
               
                 TABLE 15 
               
               
                   
               
               
                   
                   
                 SEQ 
               
               
                   
                   
                 ID 
               
               
                 Gene 
                 Nucleotide Sequence (5′-3′) 
                 NO. 
               
               
                   
               
             
            
               
                 Bcl-x 
                 TGGTTCTTACCCAGCCGCCG 
                 264 
               
               
                   
               
               
                 β-globin 623 
                 GTTATTCTTTAGAATGGTGC 
                 265 
               
               
                   
               
               
                 β-globin 654 
                 TGCTATTACCTTAACCCAGA 
                 266 
               
               
                   
               
               
                 c-myc 
                 CTGTGCTTACCGGGTTTTCCACCTCCC 
                 267 
               
               
                   
               
               
                 c-myc 
                 ATCGTCGTGACTGTCTGTTGGAGGG 
                 268 
               
               
                   
               
               
                 c-myc 
                 GCTCACGTTGAGGGGCATCG 
                 269 
               
               
                   
               
               
                 c-myc 
                 ACGTTGAGGGGCATCGTCGC 
                 270 
               
               
                   
               
               
                 c-myc 
                 GGGGCAUCGUCGUGACUGU/CUGUUGGAGGG 
                 271 
               
               
                   
               
               
                 c-myc 
                 CGUCGUGACUGUCUGUUGGAGG 
                 272 
               
               
                   
               
               
                 c-myc 
                 CGTCGTGACTGTCTGTTGGAGG 
                 273 
               
               
                   
               
               
                 c-myc 
                 GGCAUCGUCGCGGGAGGCUGCUGGAGCG 
                 274 
               
               
                   
               
               
                 c-myc 
                 CCGCGACAUAGGACGGAGAGCAGAGCCC 
                 275 
               
               
                   
               
               
                 c-myc 
                 ACTGTGAGGGCGATCGCTGC 
                 276 
               
               
                   
               
               
                 c-myc 
                 ACGATGAGTGGCATAGTCGC 
                 277 
               
               
                   
               
               
                 c-myc 
                 GGCATCGTCGCGGGAGGCTG 
                 278 
               
               
                   
               
               
                 c-myc 
                 GGGCATCGTCGCGGGAGGCT 
                 279 
               
               
                   
               
               
                 c-myc 
                 GGGGCATCGTCGCGGGAGGC 
                 280 
               
               
                   
               
               
                 c-myc 
                 AGGGGCATCGTCGCGGGAGG 
                 281 
               
               
                   
               
               
                 c-myc 
                 GAGGGGCATCGTCGCGGGAG 
                 282 
               
               
                   
               
               
                 c-myc 
                 TGAGGGGCATCGTCGCGGGA 
                 283 
               
               
                   
               
               
                 c-myc 
                 TTGAGGGGCATCGTCGCGGG 
                 284 
               
               
                   
               
               
                 c-myc 
                 GTTGAGGGGCATCGTCGCGG 
                 285 
               
               
                   
               
               
                 c-myc 
                 CGTTGAGGGGCATCGTCGCG 
                 286 
               
               
                   
               
               
                 c-myc 
                 ACGTTGAGGGGCATCGTCGC 
                 287 
               
               
                   
               
               
                 c-myc 
                 AACGTTGAGGGGCATCGTCG 
                 288 
               
               
                   
               
               
                 c-myc 
                 TAACGTTGAGGGGCATCGTC 
                 289 
               
               
                   
               
               
                 c-myc 
                 CTAACGTTGAGGGGCATCGT 
                 290 
               
               
                   
               
               
                 c-myc 
                 GCTAACGTTGAGGGGCATCG 
                 291 
               
               
                   
               
               
                 c-myc 
                 AGCTAACGTTGAGGGGCATC 
                 292 
               
               
                   
               
               
                 c-myc 
                 AAGCTAACGTTGAGGGGCAT 
                 293 
               
               
                   
               
               
                 c-myc 
                 GAAGCTAACGTTGAGGGGCA 
                 294 
               
               
                   
               
               
                 BCL-2 (rat) 
                 CTCCGCAATGCTGAAAGGTG 
                 295 
               
               
                   
               
               
                 PCNA-1 (rat) 
                 GGCGUGCCUCAAACAUGGUGGCGG 
                 296 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
               
             
               
                 TABLE 16 
               
               
                   
               
               
                   
                   
                   
                 SEQ 
               
               
                   
                 Target 
                   
                 ID 
               
               
                 Gene 
                 Location 
                 Nucleotide Sequence (5′-3′) 
                 NO. 
               
               
                   
               
             
            
               
                 Rat c-myc 
                  2553-79 
                 CTGTGCTTACCGGGTTTTCCACCTCCC 
                 297 
               
               
                   
               
               
                 Rat c-myc 
                  4140-64 
                 ATCGTCGTGACTGTCTGTTGGAGGG 
                 298 
               
               
                   
               
               
                 Rat c-myc 
                  4161-80 
                 GCTCACGTTGAGGGGCATCG 
                 299 
               
               
                   
               
               
                 Rat CYP3A2 
                  1155-74 
                 GGTCACTCACCGGTAGAGAA 
                 300 
               
               
                   
               
               
                 Rat CYP3A2 
                  1526-45 
                 GGGTTCCAAGTCTATAAAGG 
                 301 
               
               
                   
               
               
                 Human androgen 
                    31-44 
                 TGTGTCTTTTCCAG 
                 302 
               
               
                 receptor exon 2 
                   
                   
                   
               
               
                   
               
               
                 Human androgen 
                    45-67 
                 TTTGGAGACTGCCAGGGACCATG 
                 303 
               
               
                 receptor exon 2 
                   
                   
                   
               
               
                   
               
               
                 Human androgen 
                    48-67 
                 CATGGTCCCTGGCAGTCTCC 
                 304 
               
               
                 receptor exon 2 
                   
                   
                   
               
               
                   
               
               
                 Human androgen 
                    45-80 
                 TCAATGGGCAAAACATGGTCCCTGGCAGTCTCCAAA 
                 305 
               
               
                 receptor exon 2 
                   
                   
                   
               
               
                   
               
               
                 Human androgen 
                    28-43 
                 TTTGTGTTCTCCCAG 
                 306 
               
               
                 receptor exon 3 
                   
                   
                   
               
               
                   
               
               
                 Human androgen 
                    44-66 
                 GGAAACAGAAGTACCTGTGCGCC 
                 307 
               
               
                 receptor exon 3 
                   
                   
                   
               
               
                   
               
               
                 Human androgen 
                    49-66 
                 GGCGCACAGGTACTTCTG 
                 308 
               
               
                 receptor exon 3 
                   
                   
                   
               
               
                   
               
               
                 Human androgen 
                    44-79 
                 AATCATTTCTGCTGGCGCACAGGTACTTCTGTTTCC 
                 309 
               
               
                 receptor exon 3 
                   
                   
                   
               
               
                   
               
               
                 Human HCG-β 
                  1321-38 
                 CCCCTGCAGCACGCGGGT 
                 310 
               
               
                 subunit 
                   
                   
                   
               
               
                   
               
               
                 Human HCG-β 
                  1321-57 
                 GAGGCAGGGCCGGCAGGACCCCCTGCAGCACGCGGGT 
                 311 
               
               
                 subunit 
                   
                   
                   
               
               
                   
               
               
                 Human c-myc 
                  4506-25 
                 GGCATCGTCGCGGGAGGCTG 
                 312 
               
               
                   
               
               
                 Human c-myc 
                  4507-26 
                 GGGCATCGTCGCGGGAGGCT 
                 313 
               
               
                   
               
               
                 Human c-myc 
                  4508-27 
                 GGGGCATCGTCGCGGGAGGC 
                 314 
               
               
                   
               
               
                 Human c-myc 
                  4509-28 
                 AGGGGCATCGTCGCGGGAGG 
                 315 
               
               
                   
               
               
                 Human c-myc 
                  4510-29 
                 GAGGGGCATCGTCGCGGGAG 
                 316 
               
               
                   
               
               
                 Human c-myc 
                  4511-30 
                 TGAGGGGCATCGTCGCGGGA 
                 317 
               
               
                   
               
               
                 Human c-myc 
                  4512-31 
                 TTGAGGGGCATCGTCGCGGG 
                 318 
               
               
                   
               
               
                 Human c-myc 
                  4513-32 
                 GTTGAGGGGCATCGTCGCGG 
                 319 
               
               
                   
               
               
                 Human c-myc 
                  4514-33 
                 CGTTGAGGGGCATCGTCGCG 
                 320 
               
               
                   
               
               
                 Human c-myc 
                  4515-34 
                 ACGTTGAGGGGCATCGTCGC 
                 321 
               
               
                   
               
               
                 Human c-myc 
                  4516-35 
                 AACGTTGAGGGGCATCGTCG 
                 322 
               
               
                   
               
               
                 Human c-myc 
                  4517-36 
                 TAACGTTGAGGGGCATCGTC 
                 323 
               
               
                   
               
               
                 Human c-myc 
                  4518-37 
                 CTAACGTTGAGGGGCATCGT 
                 324 
               
               
                   
               
               
                 Human c-myc 
                  4519-38 
                 GCTAACGTTGAGGGGCATCG 
                 325 
               
               
                   
               
               
                 Human c-myc 
                  4520-39 
                 AGCTAACGTTGAGGGGCATC 
                 326 
               
               
                   
               
               
                 Human c-myc 
                  4521-40 
                 AAGCTAACGTTGAGGGGCAT 
                 327 
               
               
                   
               
               
                 Human c-myc 
                  4522-41 
                 GAAGCTAACGTTGAGGGGCA 
                 328 
               
               
                   
               
               
                 Human c-myc 
                  6656-75 
                 TCCTCATCTTCTTGTTCCTC 
                 329 
               
               
                   
               
               
                 Human c-myc 
                  6656-91 
                 AACAACATCGATTTCTTCCTCATCTTCTTGTTCCTC 
                 330 
               
               
                   
               
               
                 Human p53 
                 11691-708 
                 CCCGGAAGGCAGTCTGGC 
                 331 
               
               
                   
               
               
                 Human p53 
                 11689-724 
                 TCCTCCATGGCAGTGACCCGGAAGGCAGTCTGGCTG 
                 332 
               
               
                   
               
               
                 Human abl (ds of 
                   376-94 
                 CTACTGGCCGCTGAAGGGC 
                 333 
               
               
                 bcr-abl fusion 
                   
                   
                   
               
               
                 point) 
                   
                   
                   
               
               
                   
               
               
                 Human abl (ds of 
                   374-409 
                 GCTCAAAGTCAGATGCTACTGGCCGCTGAAGGGCTT 
                 334 
               
               
                 bcr-abl fusion 
                   
                   
                   
               
               
                 point) 
                   
                   
                   
               
               
                   
               
               
                 HW-1 rev 
                  5517-43 
                 TCGTCGGTCTCTCCGCTTCTTCTTGCC 
                 335 
               
               
                   
               
               
                 HW-1 rev 
                  7885-7904 
                 CTCTGGTGGTGGGTAAGGGT 
                 336 
               
               
                   
               
               
                 HW-1 rev 
                  7885-7921 
                 CGGGTCTGTCGGGTTCCCTCTGGTGGTGGGTAAGGGT 
                 337 
               
               
                   
               
               
                 Rat c-myc 
                  4140-69 
                 GGGGCAUCGUCGUGACUGUCUGUUGGAGGG 
                 338 
               
               
                   
               
               
                 Rat c-myc 
                  4141-62 
                 CGUCGUGACUGUCUGUUGGAGG 
                 339 
               
               
                   
               
               
                 Rat c-myc 
                  4141-62 
                 CGTCGTGACTGTCTGTTGGAGG 
                 340 
               
               
                   
               
               
                 Human c-myc 
                  4498-4505 
                 GGCAUCGUCGCGGGAGGCUG/CUGGAGCG 
                 341 
               
               
                   
               
               
                 Rat c-myc 
                  4364-91 
                 CCGCGACAUAGGACGGAGAGCAGAGCCC 
                 342 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
               
             
               
                 TABLE 17 
               
               
                   
               
               
                   
                   
                 SEQ ID 
               
               
                 Target 
                 Nucleotide Sequence (5′-3′) 
                 NO. 
               
               
                   
               
             
            
               
                 Hu.DMD.Exon44.25.001 
                 CTGCAGGTAAAAGCATATGGATCAA 
                 343 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.002 
                 ATCGCCTGCAGGTAAAAGCATATGG 
                 344 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.003 
                 GTCAAATCGCCTGCAGGTAAAAGCA 
                 345 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.004 
                 GATCTGTCAAATCGCCTGCAGGTAA 
                 346 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.005 
                 CAACAGATCTGTCAAATCGCCTGCA 
                 347 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.006 
                 TTTCTCAACAGATCTGTCAAATCGC 
                 348 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.007 
                 CCATTTCTCAACAGATCTGTCAAAT 
                 349 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.008 
                 ATAATGAAAACGCCGCCATTTCTCA 
                 350 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.009 
                 AAATATCTTTATATCATAATGAAAA 
                 351 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.010 
                 TGTTAGCCACTGATTAAATATCTTT 
                 352 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.011 
                 AAACTGTTCAGCTTCTGTTAGCCAC 
                 353 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.012 
                 TTGTGTCTTTCTGAGAAACTGTTCA 
                 354 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.013 
                 CCAATTCTCAGGAATTTGTGTCTTT 
                 355 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.014 
                 GTATTTAGCATGTTCCCAATTCTCA 
                 356 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.015 
                 CTTAAGATACCATTTGTATTTAGCA 
                 357 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.016 
                 CTTACCTTAAGATACCATTTGTATT 
                 358 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.017 
                 AAAGACTTACCTTAAGATACCATTT 
                 359 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.018 
                 AAATCAAAGACTTACCTTAAGATAC 
                 360 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.019 
                 AAAACAAATCAAAGACTTACCTTAA 
                 361 
               
               
                   
               
               
                 Hu.DMD.Exon44.25.020 
                 TCGAAAAAACAAATCAAAGACTTAC 
                 362 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.001 
                 CTGTAAGATACCAAAAAGGCAAAAC 
                 363 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.002 
                 CCTGTAAGATACCAAAAAGGCAAAA 
                 364 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.002.2 
                 AGTTCCTGTAAGATACCAAAAAGGC 
                 365 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.003 
                 GAGTTCCTGTAAGATACCAAAAAGG 
                 366 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.003.2 
                 CCTGGAGTTCCTGTAAGATACCAAA 
                 367 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.004 
                 TCCTGGAGTTCCTGTAAGATACCAA 
                 368 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.004.2 
                 GCCATCCTGGAGTTCCTGTAAGATA 
                 369 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.005 
                 TGCCATCCTGGAGTTCCTGTAAGAT 
                 370 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.005.2 
                 CCAATGCCATCCTGGAGTTCCTGTA 
                 371 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.006 
                 CCCAATGCCATCCTGGAGTTCCTGT 
                 372 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.006.2 
                 GCTGCCCAATGCCATCCTGGAGTTC 
                 373 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.007 
                 CGCTGCCCAATGCCATCCTGGAGTT 
                 374 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.008 
                 AACAGTTTGCCGCTGCCCAATGCCA 
                 375 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.008.2 
                 CTGACAACAGTTTGCCGCTGCCCAA 
                 376 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.009 
                 GTTGCATTCAATGTTCTGACAACAG 
                 377 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.010 
                 GCTGAATTATTTCTTCCCCAGTTGC 
                 378 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.010.2 
                 ATTATTTCTTCCCCAGTTGCATTCA 
                 379 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.011 
                 GGCATCTGTTTTTGAGGATTGCTGA 
                 380 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.011.2 
                 TTTGAGGATTGCTGAATTATTTCTT 
                 381 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.012 
                 AATTTTTCCTGTAGAATACTGGCAT 
                 382 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.012.2 
                 ATACTGGCATCTGTTTTTGAGGATT 
                 383 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.013 
                 ACCGCAGATTCAGGCTTCCCAATTT 
                 384 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.013.2 
                 AATTTTTCCTGTAGAATACTGGCAT 
                 385 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.014 
                 CTGTTTGCAGACCTCCTGCCACCGC 
                 386 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.014.2 
                 AGATTCAGGCTTCCCAATTTTTCCT 
                 387 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.015 
                 CTCTTTTTTCTGTCTGACAGCTGTT 
                 388 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.015.2 
                 ACCTCCTGCCACCGCAGATTCAGGC 
                 389 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.016 
                 CCTACCTCTTTTTTCTGTCTGACAG 
                 390 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.016.2 
                 GACAGCTGTTTGCAGACCTCCTGCC 
                 391 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.017 
                 GTCGCCCTACCTCTTTTTTCTGTCT 
                 392 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.018 
                 GATCTGTCGCCCTACCTCTTTTTTC 
                 393 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.019 
                 TATTAGATCTGTCGCCCTACCTCTT 
                 394 
               
               
                   
               
               
                 Hu.DMD.Exon45.25.020 
                 ATTCCTATTAGATCTGTCGCCCTAC 
                 395 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.001 
                 AGATACCAAAAAGGCAAAAC 
                 396 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.002 
                 AAGATACCAAAAAGGCAAAA 
                 397 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.003 
                 CCTGTAAGATACCAAAAAGG 
                 398 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.004 
                 GAGTTCCTGTAAGATACCAA 
                 399 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.005 
                 TCCTGGAGTTCCTGTAAGAT 
                 400 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.006 
                 TGCCATCCTGGAGTTCCTGT 
                 401 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.007 
                 CCCAATGCCATCCTGGAGTT 
                 402 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.008 
                 CGCTGCCCAATGCCATCCTG 
                 403 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.009 
                 CTGACAACAGTTTGCCGCTG 
                 404 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.010 
                 GTTGCATTCAATGTTCTGAC 
                 405 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.011 
                 ATTATTTCTTCCCCAGTTGC 
                 406 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.012 
                 TTTGAGGATTGCTGAATTAT 
                 407 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.013 
                 ATACTGGCATCTGTTTTTGA 
                 408 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.014 
                 AATTTTTCCTGTAGAATACT 
                 409 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.015 
                 AGATTCAGGCTTCCCAATTT 
                 410 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.016 
                 ACCTCCTGCCACCGCAGATT 
                 411 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.017 
                 GACAGCTGTTTGCAGACCTC 
                 412 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.018 
                 CTCTTTTTTCTGTCTGACAG 
                 413 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.019 
                 CCTACCTCTTTTTTCTGTCT 
                 414 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.020 
                 GTCGCCCTACCTCTTTTTTC 
                 415 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.021 
                 GATCTGTCGCCCTACCTCTT 
                 416 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.022 
                 TATTAGATCTGTCGCCCTAC 
                 417 
               
               
                   
               
               
                 Hu.DMD.Exon45.20.023 
                 ATTCCTATTAGATCTGTCGC 
                 418 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.001 
                 GGGGGATTTGAGAAAATAAAATTAC 
                 419 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.002 
                 ATTTGAGAAAATAAAATTACCTTGA 
                 420 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.002.2 
                 CTAGCCTGGAGAAAGAAGAATAAAA 
                 421 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.003 
                 AGAAAATAAAATTACCTTGACTTGC 
                 422 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.003.2 
                 TTCTTCTAGCCTGGAGAAAGAAGAA 
                 423 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.004 
                 ATAAAATTACCTTGACTTGCTCAAG 
                 424 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.004.2 
                 TTTTGTTCTTCTAGCCTGGAGAAAG 
                 425 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.005 
                 ATTACCTTGACTTGCTCAAGCTTTT 
                 426 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.005.2 
                 TATTCTTTTGTTCTTCTAGCCTGGA 
                 427 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.006 
                 CTTGACTTGCTCAAGCTTTTCTTTT 
                 428 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.006.2 
                 CAAGATATTCTTTTGTTCTTCTAGC 
                 429 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.007 
                 CTTTTAGTTGCTGCTCTTTTCCAGG 
                 430 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.008 
                 CCAGGTTCAAGTGGGATACTAGCAA 
                 431 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.008.2 
                 ATCTCTTTGAAATTCTGACAAGATA 
                 432 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.009 
                 AGCAATGTTATCTGCTTCCTCCAAC 
                 433 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.009.2 
                 AACAAATTCATTTAAATCTCTTTGA 
                 434 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.010 
                 CCAACCATAAAACAAATTCATTTAA 
                 435 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.010.2 
                 TTCCTCCAACCATAAAACAAATTCA 
                 436 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.011 
                 TTTAAATCTCTTTGAAATTCTGACA 
                 437 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.012 
                 TGACAAGATATTCTTTTGTTCTTCT 
                 438 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.012.2 
                 TTCAAGTGGGATACTAGCAATGTTA 
                 439 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.013 
                 AGATATTCTTTTGTTCTTCTAGCCT 
                 440 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.013.2 
                 CTGCTCTTTTCCAGGTTCAAGTGGG 
                 441 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.014 
                 TTCTTTTGTTCTTCTAGCCTGGAGA 
                 442 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.014.2 
                 CTTTTCTTTTAGTTGCTGCTCTTTT 
                 443 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.015 
                 TTGTTCTTCTAGCCTGGAGAAAGAA 
                 444 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.016 
                 CTTCTAGCCTGGAGAAAGAAGAATA 
                 445 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.017 
                 AGCCTGGAGAAAGAAGAATAAAATT 
                 446 
               
               
                   
               
               
                 Hu.DMD.Exon46.25.018 
                 CTGGAGAAAGAAGAATAAAATTGTT 
                 447 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.001 
                 GAAAGAAGAATAAAATTGTT 
                 448 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.002 
                 GGAGAAAGAAGAATAAAATT 
                 449 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.003 
                 AGCCTGGAGAAAGAAGAATA 
                 450 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.004 
                 CTTCTAGCCTGGAGAAAGAA 
                 451 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.005 
                 TTGTTCTTCTAGCCTGGAGA 
                 452 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.006 
                 TTCTTTTGTTCTTCTAGCCT 
                 453 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.007 
                 TGACAAGATATTCTTTTGTT 
                 454 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.008 
                 ATCTCTTTGAAATTCTGACA 
                 455 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.009 
                 AACAAATTCATTTAAATCTC 
                 456 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.010 
                 TTCCTCCAACCATAAAACAA 
                 457 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.011 
                 AGCAATGTTATCTGCTTCCT 
                 458 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.012 
                 TTCAAGTGGGATACTAGCAA 
                 459 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.013 
                 CTGCTCTTTTCCAGGTTCAA 
                 460 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.014 
                 CTTTTCTTTTAGTTGCTGCT 
                 461 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.015 
                 CTTGACTTGCTCAAGCTTTT 
                 462 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.016 
                 ATTACCTTGACTTGCTCAAG 
                 463 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.017 
                 ATAAAATTACCTTGACTTGC 
                 464 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.018 
                 AGAAAATAAAATTACCTTGA 
                 465 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.019 
                 ATTTGAGAAAATAAAATTAC 
                 466 
               
               
                   
               
               
                 Hu.DMD.Exon46.20.020 
                 GGGGGATTTGAGAAAATAAA 
                 467 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.001 
                 CTGAAACAGACAAATGCAACAACGT 
                 468 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.002 
                 AGTAACTGAAACAGACAAATGCAAC 
                 469 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.003 
                 CCACCAGTAACTGAAACAGACAAAT 
                 470 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.004 
                 CTCTTCCACCAGTAACTGAAACAGA 
                 471 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.005 
                 GGCAACTCTTCCACCAGTAACTGAA 
                 472 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.006 
                 GCAGGGGCAACTCTTCCACCAGTAA 
                 473 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.007 
                 CTGGCGCAGGGGCAACTCTTCCACC 
                 474 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.008 
                 TTTAATTGTTTGAGAATTCCCTGGC 
                 475 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.008.2 
                 TTGTTTGAGAATTCCCTGGCGCAGG 
                 476 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.009 
                 GCACGGGTCCTCCAGTTTCATTTAA 
                 477 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.009.2 
                 TCCAGTTTCATTTAATTGTTTGAGA 
                 478 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.010 
                 GCTTATGGGAGCACTTACAAGCACG 
                 479 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.010.2 
                 TACAAGCACGGGTCCTCCAGTTTCA 
                 480 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.011 
                 AGTTTATCTTGCTCTTCTGGGCTTA 
                 481 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.012 
                 TCTGCTTGAGCTTATTTTCAAGTTT 
                 482 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.012.2 
                 ATCTTGCTCTTCTGGGCTTATGGGA 
                 483 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.013 
                 CTTTATCCACTGGAGATTTGTCTGC 
                 484 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.013.2 
                 CTTATTTTCAAGTTTATCTTGCTCT 
                 485 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.014 
                 CTAACCTTTATCCACTGGAGATTTG 
                 486 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.014.2 
                 ATTTGTCTGCTTGAGCTTATTTTCA 
                 487 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.015 
                 AATGTCTAACCTTTATCCACTGGAG 
                 488 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.016 
                 TGGTTAATGTCTAACCTTTATCCAC 
                 489 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.017 
                 AGAGATGGTTAATGTCTAACCTTTA 
                 490 
               
               
                   
               
               
                 Hu.DMD.Exon47.25.018 
                 ACGGAAGAGATGGTTAATGTCTAAC 
                 491 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.001 
                 ACAGACAAATGCAACAACGT 
                 492 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.002 
                 CTGAAACAGACAAATGCAAC 
                 493 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.003 
                 AGTAACTGAAACAGACAAAT 
                 494 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.004 
                 CCACCAGTAACTGAAACAGA 
                 495 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.005 
                 CTCTTCCACCAGTAACTGAA 
                 496 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.006 
                 GGCAACTCTTCCACCAGTAA 
                 497 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.007 
                 CTGGCGCAGGGGCAACTCTT 
                 498 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.008 
                 TTGTTTGAGAATTCCCTGGC 
                 499 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.009 
                 TCCAGTTTCATTTAATTGTT 
                 500 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.010 
                 TACAAGCACGGGTCCTCCAG 
                 501 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.011 
                 GCTTATGGGAGCACTTACAA 
                 502 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.012 
                 ATCTTGCTCTTCTGGGCTTA 
                 503 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.013 
                 CTTATTTTCAAGTTTATCTT 
                 504 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.014 
                 ATTTGTCTGCTTGAGCTTAT 
                 505 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.015 
                 CTTTATCCACTGGAGATTTG 
                 506 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.016 
                 CTAACCTTTATCCACTGGAG 
                 507 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.017 
                 AATGTCTAACCTTTATCCAC 
                 508 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.018 
                 TGGTTAATGTCTAACCTTTA 
                 509 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.019 
                 AGAGATGGTTAATGTCTAAC 
                 510 
               
               
                   
               
               
                 Hu.DMD.Exon47.20.020 
                 ACGGAAGAGATGGTTAATGT 
                 511 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.001 
                 CTGAAAGGAAAATACATTTTAAAAA 
                 512 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.002 
                 CCTGAAAGGAAAATACATTTTAAAA 
                 513 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.002.2 
                 GAAACCTGAAAGGAAAATACATTTT 
                 514 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.003 
                 GGAAACCTGAAAGGAAAATACATTT 
                 515 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.003.2 
                 CTCTGGAAACCTGAAAGGAAAATAC 
                 516 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.004 
                 GCTCTGGAAACCTGAAAGGAAAATA 
                 517 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.004.2 
                 TAAAGCTCTGGAAACCTGAAAGGAA 
                 518 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.005 
                 GTAAAGCTCTGGAAACCTGAAAGGA 
                 519 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.005.2 
                 TCAGGTAAAGCTCTGGAAACCTGAA 
                 520 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.006 
                 CTCAGGTAAAGCTCTGGAAACCTGA 
                 521 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.006.2 
                 GTTTCTCAGGTAAAGCTCTGGAAAC 
                 522 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.007 
                 TGTTTCTCAGGTAAAGCTCTGGAAA 
                 523 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.007.2 
                 AATTTCTCCTTGTTTCTCAGGTAAA 
                 524 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.008 
                 TTTGAGCTTCAATTTCTCCTTGTTT 
                 525 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.008 
                 TTTTATTTGAGCTTCAATTTCTCCT 
                 526 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.009 
                 AAGCTGCCCAAGGTCTTTTATTTGA 
                 527 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.010 
                 AGGTCTTCAAGCTTTTTTTCAAGCT 
                 528 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.010.2 
                 TTCAAGCTTTTTTTCAAGCTGCCCA 
                 529 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.011 
                 GATGATTTAACTGCTCTTCAAGGTC 
                 530 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.011.2 
                 CTGCTCTTCAAGGTCTTCAAGCTTT 
                 531 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.012 
                 AGGAGATAACCACAGCAGCAGATGA 
                 532 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.012.2 
                 CAGCAGATGATTTAACTGCTCTTCA 
                 533 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.013 
                 ATTTCCAACTGATTCCTAATAGGAG 
                 534 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.014 
                 CTTGGTTTGGTTGGTTATAAATTTC 
                 535 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.014.2 
                 CAACTGATTCCTAATAGGAGATAAC 
                 536 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.015 
                 CTTAACGTCAAATGGTCCTTCTTGG 
                 537 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.015.2 
                 TTGGTTATAAATTTCCAACTGATTC 
                 538 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.016 
                 CCTACCTTAACGTCAAATGGTCCTT 
                 539 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.016.2 
                 TCCTTCTTGGTTTGGTTGGTTATAA 
                 540 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.017 
                 AGTTCCCTACCTTAACGTCAAATGG 
                 541 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.018 
                 CAAAAAGTTCCCTACCTTAACGTCA 
                 542 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.019 
                 TAAAGCAAAAAGTTCCCTACCTTAA 
                 543 
               
               
                   
               
               
                 Hu.DMD.Exon48.25.020 
                 ATATTTAAAGCAAAAAGTTCCCTAC 
                 544 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.001 
                 AGGAAAATACATTTTAAAAA 
                 545 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.002 
                 AAGGAAAATACATTTTAAAA 
                 546 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.003 
                 CCTGAAAGGAAAATACATTT 
                 547 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.004 
                 GGAAACCTGAAAGGAAAATA 
                 548 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.005 
                 GCTCTGGAAACCTGAAAGGA 
                 549 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.006 
                 GTAAAGCTCTGGAAACCTGA 
                 550 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.007 
                 CTCAGGTAAAGCTCTGGAAA 
                 551 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.008 
                 AATTTCTCCTTGTTTCTCAG 
                 552 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.009 
                 TTTTATTTGAGCTTCAATTT 
                 553 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.010 
                 AAGCTGCCCAAGGTCTTTTA 
                 554 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.011 
                 TTCAAGCTTTTTTTCAAGCT 
                 555 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.012 
                 CTGCTCTTCAAGGTCTTCAA 
                 556 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.013 
                 CAGCAGATGATTTAACTGCT 
                 557 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.014 
                 AGGAGATAACCACAGCAGCA 
                 558 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.015 
                 CAACTGATTCCTAATAGGAG 
                 559 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.016 
                 TTGGTTATAAATTTCCAACT 
                 560 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.017 
                 TCCTTCTTGGTTTGGTTGGT 
                 561 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.018 
                 CTTAACGTCAAATGGTCCTT 
                 562 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.019 
                 CCTACCTTAACGTCAAATGG 
                 563 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.020 
                 AGTTCCCTACCTTAACGTCA 
                 564 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.021 
                 CAAAAAGTTCCCTACCTTAA 
                 565 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.022 
                 TAAAGCAAAAAGTTCCCTAC 
                 566 
               
               
                   
               
               
                 Hu.DMD.Exon48.20.023 
                 ATATTTAAAGCAAAAAGTTC 
                 567 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.001 
                 CTGGGGAAAAGAACCCATATAGTGC 
                 568 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.002 
                 TCCTGGGGAAAAGAACCCATATAGT 
                 569 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.002.2 
                 GTTTCCTGGGGAAAAGAACCCATAT 
                 570 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.003 
                 CAGTTTCCTGGGGAAAAGAACCCAT 
                 571 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.003.2 
                 TTTCAGTTTCCTGGGGAAAAGAACC 
                 572 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.004 
                 TATTTCAGTTTCCTGGGGAAAAGAA 
                 573 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.004.2 
                 TGCTATTTCAGTTTCCTGGGGAAAA 
                 574 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.005 
                 ACTGCTATTTCAGTTTCCTGGGGAA 
                 575 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.005.2 
                 TGAACTGCTATTTCAGTTTCCTGGG 
                 576 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.006 
                 CTTGAACTGCTATTTCAGTTTCCTG 
                 577 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.006.2 
                 TAGCTTGAACTGCTATTTCAGTTTC 
                 578 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.007 
                 TTTAGCTTGAACTGCTATTTCAGTT 
                 579 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.008 
                 TTCCACATCCGGTTGTTTAGCTTGA 
                 580 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.009 
                 TGCCCTTTAGACAAAATCTCTTCCA 
                 581 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.009.2 
                 TTTAGACAAAATCTCTTCCACATCC 
                 582 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.010 
                 GTTTTTCCTTGTACAAATGCTGCCC 
                 583 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.010.2 
                 GTACAAATGCTGCCCTTTAGACAAA 
                 584 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.011 
                 CTTCACTGGCTGAGTGGCTGGTTTT 
                 585 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.011.2 
                 GGCTGGTTTTTCCTTGTACAAATGC 
                 586 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.012 
                 ATTACCTTCACTGGCTGAGTGGCTG 
                 587 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.013 
                 GCTTCATTACCTTCACTGGCTGAGT 
                 588 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.014 
                 AGGTTGCTTCATTACCTTCACTGGC 
                 589 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.015 
                 GCTAGAGGTTGCTTCATTACCTTCA 
                 590 
               
               
                   
               
               
                 Hu.DMD.Exon49.25.016 
                 ATATTGCTAGAGGTTGCTTCATTAC 
                 591 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.001 
                 GAAAAGAACCCATATAGTGC 
                 592 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.002 
                 GGGAAAAGAACCCATATAGT 
                 593 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.003 
                 TCCTGGGGAAAAGAACCCAT 
                 594 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.004 
                 CAGTTTCCTGGGGAAAAGAA 
                 595 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.005 
                 TATTTCAGTTTCCTGGGGAA 
                 596 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.006 
                 ACTGCTATTTCAGTTTCCTG 
                 597 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.007 
                 CTTGAACTGCTATTTCAGTT 
                 598 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.008 
                 TTTAGCTTGAACTGCTATTT 
                 599 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.009 
                 TTCCACATCCGGTTGTTTAG 
                 600 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.010 
                 TTTAGACAAAATCTCTTCCA 
                 601 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.011 
                 GTACAAATGCTGCCCTTTAG 
                 602 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.012 
                 GGCTGGTTTTTCCTTGTACA 
                 603 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.013 
                 CTTCACTGGCTGAGTGGCTG 
                 604 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.014 
                 ATTACCTTCACTGGCTGAGT 
                 605 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.015 
                 GCTTCATTACCTTCACTGGC 
                 606 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.016 
                 AGGTTGCTTCATTACCTTCA 
                 607 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.017 
                 GCTAGAGGTTGCTTCATTAC 
                 608 
               
               
                   
               
               
                 Hu.DMD.Exon49.20.018 
                 ATATTGCTAGAGGTTGCTTC 
                 609 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.001 
                 CTTTAACAGAAAAGCATACACATTA 
                 610 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.002 
                 TCCTCTTTAACAGAAAAGCATACAC 
                 611 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.002.2 
                 TTCCTCTTTAACAGAAAAGCATACA 
                 612 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.003 
                 TAACTTCCTCTTTAACAGAAAAGCA 
                 613 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.003.2 
                 CTAACTTCCTCTTTAACAGAAAAGC 
                 614 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.004 
                 TCTTCTAACTTCCTCTTTAACAGAA 
                 615 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.004.2 
                 ATCTTCTAACTTCCTCTTTAACAGA 
                 616 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.005 
                 TCAGATCTTCTAACTTCCTCTTTAA 
                 617 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.005.2 
                 CTCAGATCTTCTAACTTCCTCTTTA 
                 618 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.006 
                 AGAGCTCAGATCTTCTAACTTCCTC 
                 619 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.006.2 
                 CAGAGCTCAGATCTTCTAACTTCCT 
                 620 
               
               
                 NG-08-0731 
                   
                   
               
               
                   
               
               
                 Hu.DMD.Exon50.25.007 
                 CACTCAGAGCTCAGATCTTCTACT 
                 621 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.007.2 
                 CCTTCCACTCAGAGCTCAGATCTTC 
                 622 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.008 
                 GTAAACGGTTTACCGCCTTCCACTC 
                 623 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.009 
                 CTTTGCCCTCAGCTCTTGAAGTAAA 
                 624 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.009.2 
                 CCCTCAGCTCTTGAAGTAAACGGTT 
                 625 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.010 
                 CCAGGAGCTAGGTCAGGCTGCTTTG 
                 626 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.010.2 
                 GGTCAGGCTGCTTTGCCCTCAGCTC 
                 627 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.011 
                 AGGCTCCAATAGTGGTCAGTCCAGG 
                 628 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.011.2 
                 TCAGTCCAGGAGCTAGGTCAGGCTG 
                 629 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.012 
                 CTTACAGGCTCCAATAGTGGTCAGT 
                 630 
               
               
                 AVI-5038 
                   
                   
               
               
                   
               
               
                 Hu.DMD.Exon50.25.013 
                 GTATACTTACAGGCTCCAATAGTGG 
                 631 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.014 
                 ATCCAGTATACTTACAGGCTCCAAT 
                 632 
               
               
                   
               
               
                 Hu.DMD.Exon50.25.015 
                 ATGGGATCCAGTATACTTACAGGCT 
                 633 
               
               
                 NG-08-0741 
                   
                   
               
               
                   
               
               
                 Hu.DMD.Exon50.25.016 
                 AGAGAATGGGATCCAGTATACTTAC 
                 634 
               
               
                 NG-08-0742 
                   
                   
               
               
                   
               
               
                 Hu.DMD.Exon50.20.001 
                 ACAGAAAAGCATACACATTA 
                 635 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.002 
                 TTTAACAGAAAAGCATACAC 
                 636 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.003 
                 TCCTCTTTAACAGAAAAGCA 
                 637 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.004 
                 TAACTTCCTCTTTAACAGAA 
                 638 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.005 
                 TCTTCTAACTTCCTCTTTAA 
                 639 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.006 
                 TCAGATCTTCTAACTTCCTC 
                 640 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.007 
                 CCTTCCACTCAGAGCTCAGA 
                 641 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.008 
                 GTAAACGGTTTACCGCCTTC 
                 642 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.009 
                 CCCTCAGCTCTTGAAGTAAA 
                 643 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.010 
                 GGTCAGGCTGCTTTGCCCTC 
                 644 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.011 
                 TCAGTCCAGGAGCTAGGTCA 
                 645 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.012 
                 AGGCTCCAATAGTGGTCAGT 
                 646 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.013 
                 CTTACAGGCTCCAATAGTGG 
                 647 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.014 
                 GTATACTTACAGGCTCCAAT 
                 648 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.015 
                 ATCCAGTATACTTACAGGCT 
                 649 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.016 
                 ATGGGATCCAGTATACTTAC 
                 650 
               
               
                   
               
               
                 Hu.DMD.Exon50.20.017 
                 AGAGAATGGGATCCAGTATA 
                 651 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.001-44 
                 CTAAAATATTTTGGGTTTTTGCAAAA 
                 652 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.002-45 
                 GCTAAAATATTTTGGGTTTTTGCAAA 
                 653 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.002.2-46 
                 TAGGAGCTAAAATATTTTGGGTTTTT 
                 654 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.003 
                 AGTAGGAGCTAAAATATTTTGGGTT 
                 655 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.003.2 
                 TGAGTAGGAGCTAAAATATTTTGGG 
                 656 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.004 
                 CTGAGTAGGAGCTAAAATATTTTGGG 
                 657 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.004.2 
                 CAGTCTGAGTAGGAGCTAAAATATT 
                 658 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.005 
                 ACAGTCTGAGTAGGAGCTAAAATATT 
                 659 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.005.2 
                 GAGTAACAGTCTGAGTAGGAGCTAAA 
                 660 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.006 
                 CAGAGTAACAGTCTGAGTAGGAGCT 
                 661 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.006.2 
                 CACCAGAGTAACAGTCTGAGTAGGAG 
                 662 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.007 
                 GTCACCAGAGTAACAGTCTGAGTAG 
                 663 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.007.2 
                 AACCACAGGTTGTGTCACCAGAGTAA 
                 664 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.008 
                 GTTGTGTCACCAGAGTAACAGTCTG 
                 665 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.009 
                 TGGCAGTTTCCTTAGTAACCACAGGT 
                 666 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.010 
                 ATTTCTAGTTTGGAGATGGCAGTTTC 
                 667 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.010.2 
                 GGAAGATGGCATTTCTAGTTTGGAG 
                 668 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.011 
                 CATCAAGGAAGATGGCATTTCTAGTT 
                 669 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.011.2 
                 GAGCAGGTACCTCCAACATCAAGGAA 
                 670 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.012 
                 ATCTGCCAGAGCAGGTACCTCCAAC 
                 671 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.013 
                 AAGTTCTGTCCAAGCCCGGTTGAAAT 
                 672 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.013.2 
                 CGGTTGAAATCTGCCAGAGCAGGTAC 
                 673 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.014 
                 GAGAAAGCCAGTCGGTAAGTTCTGTC 
                 674 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.014.2 
                 GTCGGTAAGTTCTGTCCAAGCCCGG 
                 675 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.015 
                 ATAACTTGATCAAGCAGAGAAAGCCA 
                 676 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.015.2 
                 AAGCAGAGAAAGCCAGTCGGTAAGT 
                 677 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.016 
                 CACCCTCTGTGATTTTATAACTTGAT 
                 678 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.017 
                 CAAGGTCACCCACCATCACCCTCTGT 
                 679 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.017.2 
                 CATCACCCTCTGTGATTTTATAACT 
                 680 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.018 
                 CTTCTGCTTGATGATCATCTCGTTGA 
                 681 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.019 
                 CCTTCTGCTTGATGATCATCTCGTTG 
                 682 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.019.2 
                 ATCTCGTTGATATCCTCAAGGTCACC 
                 683 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.020 
                 TCATACCTTCTGCTTGATGATCATCT 
                 684 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.020.2 
                 TCATTTTTTCTCATACCTTCTGCTTG 
                 685 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.021 
                 TTTTCTCATACCTTCTGCTTGATGAT 
                 686 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.022 
                 TTTTATCATTTTTTCTCATACCTTCT 
                 687 
               
               
                   
               
               
                 Hu.DMD.Exon51.25.023 
                 CCAACTTTTATCATTTTTTCTCATAC 
                 688 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.001 
                 ATATTTTGGGTTTTTGCAAA 
                 689 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.002 
                 AAAATATTTTGGGTTTTTGC 
                 690 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.003 
                 GAGCTAAAATATTTTGGGTT 
                 691 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.004 
                 AGTAGGAGCTAAAATATTTT 
                 692 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.005 
                 GTCTGAGTAGGAGCTAAAAT 
                 693 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.006 
                 TAACAGTCTGAGTAGGAGCT 
                 694 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.007 
                 CAGAGTAACAGTCTGAGTAG 
                 695 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.008 
                 CACAGGTTGTGTCACCAGAG 
                 696 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.009 
                 AGTTTCCTTAGTAACCACAG 
                 697 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.010 
                 TAGTTTGGAGATGGCAGTTT 
                 698 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.011 
                 GGAAGATGGCATTTCTAGTT 
                 699 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.012 
                 TACCTCCAACATCAAGGAAG 
                 700 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.013 
                 ATCTGCCAGAGCAGGTACCT 
                 701 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.014 
                 CCAAGCCCGGTTGAAATCTG 
                 702 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.015 
                 GTCGGTAAGTTCTGTCCAAG 
                 703 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.016 
                 AAGCAGAGAAAGCCAGTCGG 
                 704 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.017 
                 TTTTATAACTTGATCAAGCA 
                 705 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.018 
                 CATCACCCTCTGTGATTTTA 
                 706 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.019 
                 CTCAAGGTCACCCACCATCA 
                 707 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.020 
                 CATCTCGTTGATATCCTCAA 
                 708 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.021 
                 CTTCTGCTTGATGATCATCT 
                 709 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.022 
                 CATACCTTCTGCTTGATGAT 
                 710 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.023 
                 TTTCTCATACCTTCTGCTTG 
                 711 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.024 
                 CATTTTTTCTCATACCTTCT 
                 712 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.025 
                 TTTATCATTTTTTCTCATAC 
                 713 
               
               
                   
               
               
                 Hu.DMD.Exon51.20.026 
                 CAACTTTTATCATTTTTTCT 
                 714 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.001 
                 CTGTAAGAACAAATATCCCTTAGTA 
                 715 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.002 
                 TGCCTGTAAGAACAAATATCCCTTA 
                 716 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.002.2 
                 GTTGCCTGTAAGAACAAATATCCCT 
                 717 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.003 
                 ATTGTTGCCTGTAAGAACAAATATC 
                 718 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.003.2 
                 GCATTGTTGCCTGTAAGAACAAATA 
                 719 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.004 
                 CCTGCATTGTTGCCTGTAAGAACAA 
                 720 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.004.2 
                 ATCCTGCATTGTTGCCTGTAAGAAC 
                 721 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.005 
                 CAAATCCTGCATTGTTGCCTGTAAG 
                 722 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.005.2 
                 TCCAAATCCTGCATTGTTGCCTGTA 
                 723 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.006 
                 TGTTCCAAATCCTGCATTGTTGCCT 
                 724 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.006.2 
                 TCTGTTCCAAATCCTGCATTGTTGC 
                 725 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.007 
                 AACTGGGGACGCCTCTGTTCCAAAT 
                 726 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.007.2 
                 GCCTCTGTTCCAAATCCTGCATTGT 
                 727 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.008 
                 CAGCGGTAATGAGTTCTTCCAACTG 
                 728 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.008.2 
                 CTTCCAACTGGGGACGCCTCTGTTC 
                 729 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.009 
                 CTTGTTTTTCAAATTTTGGGCAGCG 
                 730 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.010 
                 CTAGCCTCTTGATTGCTGGTCTTGT 
                 731 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.010.2 
                 TTTTCAAATTTTGGGCAGCGGTAAT 
                 732 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.011 
                 TTCGATCCGTAATGATTGTTCTAGC 
                 733 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.011.2 
                 GATTGCTGGTCTTGTTTTTCAAATT 
                 734 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.012 
                 CTTACTTCGATCCGTAATGATTGTT 
                 735 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.012.2 
                 TTGTTCTAGCCTCTTGATTGCTGGT 
                 736 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.013 
                 AAAAACTTACTTCGATCCGTAATGA 
                 737 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.014 
                 TGTTAAAAAACTTACTTCGATCCGT 
                 738 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.015 
                 ATGCTTGTTAAAAAACTTACTTCGA 
                 739 
               
               
                   
               
               
                 Hu.DMD.Exon52.25.016 
                 GTCCCATGCTTGTTAAAAAACTTAC 
                 740 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.001 
                 AGAACAAATATCCCTTAGTA 
                 741 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.002 
                 GTAAGAACAAATATCCCTTA 
                 742 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.003 
                 TGCCTGTAAGAACAAATATC 
                 743 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.004 
                 ATTGTTGCCTGTAAGAACAA 
                 744 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.005 
                 CCTGCATTGTTGCCTGTAAG 
                 745 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.006 
                 CAAATCCTGCATTGTTGCCT 
                 746 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.007 
                 GCCTCTGTTCCAAATCCTGC 
                 747 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.008 
                 CTTCCAACTGGGGACGCCTC 
                 748 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.009 
                 CAGCGGTAATGAGTTCTTCC 
                 749 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.010 
                 TTTTCAAATTTTGGGCAGCG 
                 750 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.011 
                 GATTGCTGGTCTTGTTTTTC 
                 751 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.012 
                 TTGTTCTAGCCTCTTGATTG 
                 752 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.013 
                 TTCGATCCGTAATGATTGTT 
                 753 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.014 
                 CTTACTTCGATCCGTAATGA 
                 754 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.015 
                 AAAAACTTACTTCGATCCGT 
                 755 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.016 
                 TGTTAAAAAACTTACTTCGA 
                 756 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.017 
                 ATGCTTGTTAAAAAACTTAC 
                 757 
               
               
                   
               
               
                 Hu.DMD.Exon52.20.018 
                 GTCCCATGCTTGTTAAAAAA 
                 758 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.001 
                 CTAGAATAAAAGGAAAAATAAATAT 
                 759 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.002 
                 AACTAGAATAAAAGGAAAAATAAAT 
                 760 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.002.2 
                 TTCAACTAGAATAAAAGGAAAAATA 
                 761 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.003 
                 CTTTCAACTAGAATAAAAGGAAAAA 
                 762 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.003.2 
                 ATTCTTTCAACTAGAATAAAAGGAA 
                 763 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.004 
                 GAATTCTTTCAACTAGAATAAAAGG 
                 764 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.004.2 
                 TCTGAATTCTTTCAACTAGAATAAA 
                 765 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.005 
                 ATTCTGAATTCTTTCAACTAGAATA 
                 766 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.005.2 
                 CTGATTCTGAATTCTTTCAACTAGA 
                 767 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.006 
                 CACTGATTCTGAATTCTTTCAACTA 
                 768 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.006.2 
                 TCCCACTGATTCTGAATTCTTTCAA 
                 769 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.007 
                 CATCCCACTGATTCTGAATTCTTTC 
                 770 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.008 
                 TACTTCATCCCACTGATTCTGAATT 
                 771 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.008.2 
                 CTGAAGGTGTTCTTGTACTTCATCC 
                 772 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.009 
                 CGGTTCTGAAGGTGTTCTTGTACT 
                 773 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.009.2 
                 CTGTTGCCTCCGGTTCTGAAGGTGT 
                 774 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.010 
                 TTTCATTCAACTGTTGCCTCCGGTT 
                 775 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.010.2 
                 TAACATTTCATTCAACTGTTGCCTC 
                 776 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.011 
                 TTGTGTTGAATCCTTTAACATTTCA 
                 777 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.012 
                 TCTTCCTTAGCTTCCAGCCATTGTG 
                 778 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.012.2 
                 CTTAGCTTCCAGCCATTGTGTTGAA 
                 779 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.013 
                 GTCCTAAGACCTGCTCAGCTTCTTC 
                 780 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.013.2 
                 CTGCTCAGCTTCTTCCTTAGCTTCC 
                 781 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.014 
                 CTCAAGCTTGGCTCTGGCCTGTCCT 
                 782 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.014.2 
                 GGCCTGTCCTAAGACCTGCTCAGCT 
                 783 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.015 
                 TAGGGACCCTCCTTCCATGACTCAA 
                 784 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.016 
                 TTTGGATTGCATCTACTGTATAGGG 
                 785 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.016.2 
                 ACCCTCCTTCCATGACTCAAGCTTG 
                 786 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.017 
                 CTTGGTTTCTGTGATTTTCTTTTGG 
                 787 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.017.2 
                 ATCTACTGTATAGGGACCCTCCTTC 
                 788 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.018 
                 CTAACCTTGGTTTCTGTGATTTTCT 
                 789 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.018.2 
                 TTTCTTTTGGATTGCATCTACTGTA 
                 790 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.019 
                 TGATACTAACCTTGGTTTCTGTGAT 
                 791 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.020 
                 ATCTTTGATACTAACCTTGGTTTCT 
                 792 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.021 
                 AAGGTATCTTTGATACTAACCTTGG 
                 793 
               
               
                   
               
               
                 Hu.DMD.Exon53.25.022 
                 TTAAAAAGGTATCTTTGATACTAAC 
                 794 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.001 
                 ATAAAAGGAAAAATAAATAT 
                 795 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.002 
                 GAATAAAAGGAAAAATAAAT 
                 796 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.003 
                 AACTAGAATAAAAGGAAAAA 
                 797 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.004 
                 CTTTCAACTAGAATAAAAGG 
                 798 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.005 
                 GAATTCTTTCAACTAGAATA 
                 799 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.006 
                 ATTCTGAATTCTTTCAACTA 
                 800 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.007 
                 TACTTCATCCCACTGATTCT 
                 801 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.008 
                 CTGAAGGTGTTCTTGTACT 
                 802 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.009 
                 CTGTTGCCTCCGGTTCTGAA 
                 803 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.010 
                 TAACATTTCATTCAACTGTT 
                 804 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.011 
                 TTGTGTTGAATCCTTTAACA 
                 805 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.012 
                 CTTAGCTTCCAGCCATTGTG 
                 806 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.013 
                 CTGCTCAGCTTCTTCCTTAG 
                 807 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.014 
                 GGCCTGTCCTAAGACCTGCT 
                 808 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.015 
                 CTCAAGCTTGGCTCTGGCCT 
                 809 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.016 
                 ACCCTCCTTCCATGACTCAA 
                 810 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.017 
                 ATCTACTGTATAGGGACCCT 
                 811 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.018 
                 TTTCTTTTGGATTGCATCTA 
                 812 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.019 
                 CTTGGTTTCTGTGATTTTCT 
                 813 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.020 
                 CTAACCTTGGTTTCTGTGAT 
                 814 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.021 
                 TGATACTAACCTTGGTTTCT 
                 815 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.022 
                 ATCTTTGATACTAACCTTGG 
                 816 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.023 
                 AAGGTATCTTTGATACTAAC 
                 817 
               
               
                   
               
               
                 Hu.DMD.Exon53.20.024 
                 TTAAAAAGGTATCTTTGATA 
                 818 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.001 
                 CTATAGATTTTTATGAGAAAGAGA 
                 819 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.002 
                 AACTGCTATAGATTTTTATGAGAAA 
                 820 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.003 
                 TGGCCAACTGCTATAGATTTTTATG 
                 821 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.004 
                 GTCTTTGGCCAACTGCTATAGATTT 
                 822 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.005 
                 CGGAGGTCTTTGGCCAACTGCTATA 
                 823 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.006 
                 ACTGGCGGAGGTCTTTGGCCAACTG 
                 824 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.007 
                 TTTGTCTGCCACTGGCGGAGGTCTT 
                 825 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.008 
                 AGTCATTTGCCACATCTACATTTGT 
                 826 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.008.2 
                 TTTGCCACATCTACATTTGTCTGCC 
                 827 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.009 
                 CCGGAGAAGTTTCAGGGCCAAGTCA 
                 828 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.010 
                 GTATCATCTGCAGAATAATCCCGGA 
                 829 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.010.2 
                 TAATCCCGGAGAAGTTTCAGGGCCA 
                 830 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.011 
                 TTATCATGTGGACTTTTCTGGTATC 
                 831 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.012 
                 AGAGGCATTGATATTCTCTGTTATC 
                 832 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.012.2 
                 ATGTGGACTTTTCTGGTATCATCTG 
                 833 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.013 
                 CTTTTATGAATGCTTCTCCAAGAGG 
                 834 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.013.2 
                 ATATTCTCTGTTATCATGTGGACTT 
                 835 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.014 
                 CATACCTTTTATGAATGCTTCTCCA 
                 836 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.014.2 
                 CTCCAAGAGGCATTGATATTCTCTG 
                 837 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.015 
                 TAATTCATACCTTTTATGAATGCTT 
                 838 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.015.2 
                 CTTTTATGAATGCTTCTCCAAGAGG 
                 839 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.016 
                 TAATGTAATTCATACCTTTTATGAA 
                 840 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.017 
                 AGAAATAATGTAATTCATACCTTTT 
                 841 
               
               
                   
               
               
                 Hu.DMD.Exon54.25.018 
                 GTTTTAGAAATAATGTAATTCATAC 
                 842 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.001 
                 GATTTTTATGAGAAAGAGA 
                 843 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.002 
                 CTATAGATTTTTATGAGAAA 
                 844 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.003 
                 AACTGCTATAGATTTTTATG 
                 845 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.004 
                 TGGCCAACTGCTATAGATTT 
                 846 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.005 
                 GTCTTTGGCCAACTGCTATA 
                 847 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.006 
                 CGGAGGTCTTTGGCCAACTG 
                 848 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.007 
                 TTTGTCTGCCACTGGCGGAG 
                 849 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.008 
                 TTTGCCACATCTACATTTGT 
                 850 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.009 
                 TTCAGGGCCAAGTCATTTGC 
                 851 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.010 
                 TAATCCCGGAGAAGTTTCAG 
                 852 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.011 
                 GTATCATCTGCAGAATAATC 
                 853 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.012 
                 ATGTGGACTTTTCTGGTATC 
                 854 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.013 
                 ATATTCTCTGTTATCATGTG 
                 855 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.014 
                 CTCCAAGAGGCATTGATATT 
                 856 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.015 
                 CTTTTATGAATGCTTCTCCA 
                 857 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.016 
                 CATACCTTTTATGAATGCTT 
                 858 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.017 
                 TAATTCATACCTTTTATGAA 
                 859 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.018 
                 TAATGTAATTCATACCTTTT 
                 860 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.019 
                 AGAAATAATGTAATTCATAC 
                 861 
               
               
                   
               
               
                 Hu.DMD.Exon54.20.020 
                 GTTTTAGAAATAATGTAATT 
                 862 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.001 
                 CTGCAAAGGACCAAATGTTCAGATG 
                 863 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.002 
                 TCACCCTGCAAAGGACCAAATGTTC 
                 864 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.003 
                 CTCACTCACCCTGCAAAGGACCAAA 
                 865 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.004 
                 TCTCGCTCACTCACCCTGCAAAGGA 
                 866 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.005 
                 CAGCCTCTCGCTCACTCACCCTGCA 
                 867 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.006 
                 CAAAGCAGCCTCTCGCTCACTCACC 
                 868 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.007 
                 TCTTCCAAAGCAGCCTCTCGCTCAC 
                 869 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.007.2 
                 TCTATGAGTTTCTTCCAAAGCAGCC 
                 870 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.008 
                 GTTGCAGTAATCTATGAGTTTCTTC 
                 871 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.008.2 
                 GAACTGTTGCAGTAATCTATGAGTT 
                 872 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.009 
                 TTCCAGGTCCAGGGGGAACTGTTGC 
                 873 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.010 
                 GTAAGCCAGGCAAGAAACTTTTCCA 
                 874 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.010.2 
                 CCAGGCAAGAAACTTTTCCAGGTCC 
                 875 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.011 
                 TGGCAGTTGTTTCAGCTTCTGTAAG 
                 876 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.011.2 
                 TTCAGCTTCTGTAAGCCAGGCAAGA 
                 877 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.012 
                 GGTAGCATCCTGTAGGACATTGGCA 
                 878 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.012.2 
                 GACATTGGCAGTTGTTTCAGCTTCT 
                 879 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.013 
                 TCTAGGAGCCTTTCCTTACGGGTAG 
                 880 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.014 
                 CTTTTACTCCCTTGGAGTCTTCTAG 
                 881 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.014.2 
                 GAGCCTTTCCTTACGGGTAGCATCC 
                 882 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.015 
                 TTGCCATTGTTTCATCAGCTCTTTT 
                 883 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.015.2 
                 CTTGGAGTCTTCTAGGAGCCTTTCC 
                 884 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.016 
                 CTTACTTGCCATTGTTTCATCAGCT 
                 885 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.016.2 
                 CAGCTCTTTTACTCCCTTGGAGTCT 
                 886 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.017 
                 CCTGACTTACTTGCCATTGTTTCAT 
                 887 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.018 
                 AAATGCCTGACTTACTTGCCATTGT 
                 888 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.019 
                 AGCGGAAATGCCTGACTTACTTGCC 
                 889 
               
               
                   
               
               
                 Hu.DMD.Exon55.25.020 
                 GCTAAAGCGGAAATGCCTGACTTAC 
                 890 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.001 
                 AAGGACCAAATGTTCAGATG 
                 891 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.002 
                 CTGCAAAGGACCAAATGTTC 
                 892 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.003 
                 TCACCCTGCAAAGGACCAAA 
                 893 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.004 
                 CTCACTCACCCTGCAAAGGA 
                 894 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.005 
                 TCTCGCTCACTCACCCTGCA 
                 895 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.006 
                 CAGCCTCTCGCTCACTCACC 
                 896 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.007 
                 CAAAGCAGCCTCTCGCTCAC 
                 897 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.008 
                 TCTATGAGTTTCTTCCAAAG 
                 898 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.009 
                 GAACTGTTGCAGTAATCTAT 
                 899 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.010 
                 TTCCAGGTCCAGGGGGAACT 
                 900 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.011 
                 CCAGGCAAGAAACTTTTCCA 
                 901 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.012 
                 TTCAGCTTCTGTAAGCCAGG 
                 902 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.013 
                 GACATTGGCAGTTGTTTCAG 
                 903 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.014 
                 GGTAGCATCCTGTAGGACAT 
                 904 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.015 
                 GAGCCTTTCCTTACGGGTAG 
                 905 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.016 
                 CTTGGAGTCTTCTAGGAGCC 
                 906 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.017 
                 CAGCTCTTTTACTCCCTTGG 
                 907 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.018 
                 TTGCCATTGTTTCATCAGCT 
                 908 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.019 
                 CTTACTTGCCATTGTTTCAT 
                 909 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.020 
                 CCTGACTTACTTGCCATTGT 
                 910 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.021 
                 AAATGCCTGACTTACTTGCC 
                 911 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.022 
                 AGCGGAAATGCCTGACTTAC 
                 912 
               
               
                   
               
               
                 Hu.DMD.Exon55.20.023 
                 GCTAAAGCGGAAATGCCTGA 
                 913 
               
               
                   
               
               
                 H50A(+02+30)-AVI-5656 
                 CCACTCAGAGCTCAGATCTTCTAACTTCC 
                 914 
               
               
                   
               
               
                 H50D(+07−18)-AVI-5915 
                 GGGATCCAGTATACTTACAGGCTCC 
                 915 
               
               
                   
               
               
                 H50A(+07+33) 
                 CTTCCACTCAGAGCTCAGATCTTCTAA 
                 916 
               
               
                   
               
               
                 H51A(+61+90)-AVI-4657 
                 ACATCAAGGAAGATGGCATTTCTAGTTTGG 
                 917 
               
               
                   
               
               
                 H51A(+66+95)-AVI-4658 
                 CTCCAACATCAAGGAAGATGGCATTTCTAG 
                 918 
               
               
                   
               
               
                 H51A(+111+134) 
                 TTCTGTCCAAGCCCGGTTGAAATC 
                 919 
               
               
                   
               
               
                 H51A(+175+195) 
                 CACCCACCATCACCCTCYGTG 
                 920 
               
               
                   
               
               
                 H51A(+199+220) 
                 ATCATCTCGTTGATATCCTCAA 
                 921 
               
               
                   
               
               
                 H51A(+66+90) 
                 ACATCAAGGAAGATGGCATTTCTAG 
                 922 
               
               
                   
               
               
                 H51A(−01+25) 
                 ACCAGAGTAACAGTCTGAGTAGGAGC 
                 923 
               
               
                   
               
               
                 h51AON1 
                 TCAAGGAAGATGGCATTTCT 
                 924 
               
               
                   
               
               
                 h51AON2 
                 CCTCTGTGATTTTATAACTTGAT 
                 925 
               
               
                   
               
               
                 H51D(+08−17) 
                 ATCATTTTTTCTCATACCTTCTGCT 
                 926 
               
               
                   
               
               
                 H51D(+16−07) 
                 CTCATACCTTCTGCTTGATGATC 
                 927 
               
               
                   
               
               
                 hAON#23 
                 TGGCATTTCTAGTTTGG 
                 928 
               
               
                   
               
               
                 hAON#24 
                 CCAGAGCAGGTACCTCCAACATC 
                 929 
               
               
                   
               
               
                 H44A(+61+84) 
                 TGTTCAGCTTCTGTTAGCCACTGA 
                 930 
               
               
                   
               
               
                 H44A(+85+104) 
                 TTTGTGTCTTTCTGAGAAAC 
                 931 
               
               
                   
               
               
                 h44AON1 
                 CGCCGCCATTTCTCAACAG 
                 932 
               
               
                   
               
               
                 H44A(−06+14) 
                 ATCTGTCAAATCGCCTGCAG 
                 933 
               
               
                   
               
               
                 H45A(+71+90) 
                 TGTTTTTGAGGATTGCTGAA 
                 934 
               
               
                   
               
               
                 h45AON1 
                 GCTGAATTATTTCTTCCCC 
                 935 
               
               
                   
               
               
                 h45AON5 
                 GCCCAATGCCATCCTGG 
                 936 
               
               
                   
               
               
                 H45A(−06+20) 
                 CCAATGCCATCCTGGAGTTCCTGTAA 
                 937 
               
               
                   
               
               
                 H53A(+39+69) 
                 CATTCAACTGTTGCCTCCGGTTCTGAAGGTG 
                 938 
               
               
                   
               
               
                 H53A(+23+47) 
                 CTGAAGGTGTTCTTGTACTTCATCC 
                 939 
               
               
                   
               
               
                 h53AON1 
                 CTGTTGCCTCCGGTTCTG 
                 940 
               
               
                   
               
               
                 H53A(−12+10) 
                 ATTCTTTCAACTAGAATAAAAG 
                 941 
               
               
                   
               
               
                 huEx45.30.66 
                 GCCATCCTGGAGTTCCTGTAAGATACCAAA 
                 942 
               
               
                   
               
               
                 huEx45.30.71 
                 CCAATGCCATCCTGGAGTTCCTGTAAGATA 
                 943 
               
               
                   
               
               
                 huEx45.30.79 
                 GCCGCTGCCCAATGCCATCCTGGAGTTCCT 
                 944 
               
               
                   
               
               
                 huEx45.30.83 
                 GTTTGCCGCTGCCCAATGCCATCCTGGAGT 
                 945 
               
               
                   
               
               
                 huEx45.30.88 
                 CAACAGTTTGCCGCTGCCCAATGCCATCCT 
                 946 
               
               
                   
               
               
                 huEx45.30.92 
                 CTGACAACAGTTTGCCGCTGCCCAATGCCA 
                 947 
               
               
                   
               
               
                 huEx45.30.96 
                 TGTTCTGACAACAGTTTGCCGCTGCCCAAT 
                 948 
               
               
                   
               
               
                 huEx45.30.99 
                 CAATGTTCTGACAACAGTTTGCCGCTGCCC 
                 949 
               
               
                   
               
               
                 huEx45.30.103 
                 CATTCAATGTTCTGACAACAGTTTGCCGCT 
                 950 
               
               
                   
               
               
                 huEx45.30.120 
                 TATTTCTTCCCCAGTTGCATTCAATGTTCT 
                 951 
               
               
                   
               
               
                 huEx45.30.127 
                 GCTGAATTATTTCTTCCCCAGTTGCATTCA 
                 952 
               
               
                   
               
               
                 huEx45.30.132 
                 GGATTGCTGAATTATTTCTTCCCCAGTTGC 
                 953 
               
               
                   
               
               
                 huEx45.30.137 
                 TTTGAGGATTGCTGAATTATTTCTTCCCCA 
                 954 
               
               
                   
               
               
                 huEx53.30.84 
                 GTACTTCATCCCACTGATTCTGAATTCTTT 
                 955 
               
               
                   
               
               
                 huEx53.30.88 
                 TCTTGTACTTCATCCCACTGATTCTGAATT 
                 956 
               
               
                   
               
               
                 huEx53.30.91 
                 TGTTCTTGTACTTCATCCCACTGATTCTGA 
                 957 
               
               
                   
               
               
                 huEx53.30.103 
                 CGGTTCTGAAGGTGTTCTTGTACTTCATCC 
                 958 
               
               
                   
               
               
                 huEx53.30.106 
                 CTCCGGTTCTGAAGGTGTTCTTGTACTTCA 
                 959 
               
               
                   
               
               
                 huEx53.30.109 
                 TGCCTCCGGTTCTGAAGGTGTTCTTGTACT 
                 960 
               
               
                   
               
               
                 huEx53.30.112 
                 TGTTGCCTCCGGTTCTGAAGGTGTTCTTGT 
                 961 
               
               
                   
               
               
                 huEx53.30.115 
                 AACTGTTGCCTCCGGTTCTGAAGGTGTTCT 
                 962 
               
               
                   
               
               
                 huEx53.30.118 
                 TTCAACTGTTGCCTCCGGTTCTGAAGGTGT 
                 963 
               
               
                   
               
            
           
         
       
     
     Step 1: Antibody Conjugation with Maleimide-PEG-NHS Followed by siRNA-DMD Conjugates 
     Anti-dystrophin antibody is exchanged with 1× Phosphate buffer (pH 7.4) and made up to 5 mg/ml concentration. To this solution, 2 equivalents of SMCC linker or maleimide-PEGxkDa-NHS (x=1, 5, 10, 20) is added and rotated for 4 hours at room temperature. Unreacted maleimide-PEG is removed by spin filtration using 50 kDa MWCO Amicon spin filters and PBS pH 7.4. The antibody-PEG-Mal conjugate is collected and transferred into a reaction vessel. Various siRNA conjugates are synthesized using sequences listed in Tables 13-17. siRNA-DMD conjugates (2 equivalents) is added at RT to the antibody-PEG-maleimide in PBS and rotated overnight. The reaction mixture is analyzed by analytical SAX column chromatography and conjugate along with unreacted antibody and siRNA is seen. 
     Step 2: Purification 
     The crude reaction mixture is purified by AKTA explorer FPLC using anion exchange chromatography. Fractions containing the antibody-PEG-DMD conjugate are pooled, concentrated and buffer exchanged with PBS, pH 7.4. Antibody siRNA conjugates with SMCC linker, PEG1 kDa, PEG5 kDa and PEG10 kDa are separated based on the siRNA loading. 
     Step-3: Analysis of the Purified Conjugate 
     The isolated conjugate is characterized by either mass spec or SDS-PAGE. The purity of the conjugate is assessed by analytical HPLC using anion exchange chromatography. 
     Example 8. Additional Sequences 
       
     
       
         
           
               
             
               
                 TABLE 18 
               
             
            
               
                   
               
               
                 illustrates additional polynucleic acid molecule 
               
               
                  sequences described herein. 
               
            
           
           
               
               
               
               
               
            
               
                   
                 AO name 
                 Location 
                   
                   
               
               
                   
                 (h, H: 
                 from 
                   
                 SEQ 
               
               
                   
                 Human; 
                 acceptor 
                   
                 ID 
               
               
                 Exon 
                 M: mouse) 
                 site 
                 Sequence 
                 NO: 
               
               
                   
               
            
           
           
               
               
               
               
               
            
               
                 2 
                 hEx2_Ac12 
                 12 
                 CCA UUU UGU GAA UGU UUU CUU UUG 
                 964 
               
               
                   
                   
                   
                 AAC AUC 
                   
               
               
                   
               
               
                 2 
                 hEx2_Ac19 
                 19 
                 CCC AUU UUG UGA AUG UUU UCU UUU 
                 965 
               
               
                   
               
               
                 2 
                 hEx2_Ac32 
                 32 
                 UUG UGC AUU UAC CCA UUU UGU G 
                 966 
               
               
                   
               
               
                 2 
                 hEx2_Ac35 
                 35 
                 GAA AAU UGU GCA UUU ACC CAU UUU 
                 967 
               
               
                   
               
               
                 3 
                 hEx3_Ac20 
                 20 
                 GUA GGU CAC UGA AGA GGU UCU 
                 968 
               
               
                   
               
               
                 4 
                 hEx4_Ac11 
                 11 
                 UGU UCA GGG CAU GAA CUC UUG UGG 
                 969 
               
               
                   
                   
                   
                 AUC CUU 
                   
               
               
                   
               
               
                 5 
                 hEx5_Ac25 
                 25 
                 UCA GUU UAU GAU UUC CAU CUA CGA 
                 970 
               
               
                   
                   
                   
                 UGU CAG U 
                   
               
               
                   
               
               
                 6 
                 hEx6_Ac69 
                 69 
                 UAC GAG UUG AUU GUC GGA CCC AG 
                 971 
               
               
                   
               
               
                 7 
                 hEx_Ac45 
                 45 
                 UGC AUG UUC CAG UCG UUG UGU GG 
                 972 
               
               
                   
               
               
                 8 
                 hEx8_Ac−6 
                 −6 
                 GAU AGG UGG UAU CAA CAU CUG UAA 
                 973 
               
               
                   
               
               
                 8 
                 hEx8_Ac26 
                 26 
                 CUU CCU GGA UGG CUU CAA U 
                 974 
               
               
                   
               
               
                 8 
                 hEx8_Ac84 
                 84 
                 GUA CAU UAA GAU GGA CUU C 
                 975 
               
               
                   
               
               
                 9 
                 hEx9_Ac−6 
                 −6 
                 CCC UGU GCU AGA CUG ACC GUG AUC 
                 976 
               
               
                   
                   
                   
                 UGC AG 
                   
               
               
                   
               
               
                 10 
                 hEx10_Ac−5 
                 −5 
                 CAG GAG CUU CCA AAU GCU GCA 
                 977 
               
               
                   
               
               
                 10 
                 hEx10_Ac98 
                 98 
                 UCC UCA GCA GAA AGA AGC CAC G 
                 978 
               
               
                   
               
               
                 11 
                 hEx11_Ac75 
                 75 
                 CAU CUU CUG AUA AUU UUC CUG UU 
                 979 
               
               
                   
               
               
                 12 
                 hEx12_Ac52 
                 52 
                 UCU UCU GUU UUU GUU AGC CAG UCA 
                 980 
               
               
                   
               
               
                 13 
                 hEx13_Ac77 
                 77 
                 CAG CAG UUG CGU GAU CUC CAC UAG 
                 981 
               
               
                   
               
               
                 14 
                 hEx14_Ac32 
                 32 
                 GUA AAA GAA CCC AGC GGU CUU CUG 
                 982 
               
               
                   
                   
                   
                 UCC AUC 
                   
               
               
                   
               
               
                 15 
                 hEx15_Ac48 
                 48 
                 UCU UUA AAG CCA GUU GUG UGA AUC 
                 983 
               
               
                   
               
               
                 16 
                 hEx16_Ac12 
                 12 
                 CUA GAU CCG CUU UUA AAA CCU GUU 
                 984 
               
               
                   
                   
                   
                 AAA ACA A 
                   
               
               
                   
               
               
                 16 
                 hEx16_Ac11 
                 11 
                 GAU UGC UUU UUC UUU UCU AGA UCC 
                 985 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 17 
                 hEx17_Ac−7 
                 −7 
                 UGA CAG CCU GUG AAA UCU GUG AG 
                 986 
               
               
                   
               
               
                 17 
                 hEx17_Ac36 
                 36 
                 CCA UUA CAG UUG UCU GUG UU 
                 987 
               
               
                   
               
               
                 17 
                 hEx17_Ac132 
                 132 
                 UAA UCU GCC UCU UCU UUU GG 
                 988 
               
               
                   
               
               
                 18 
                 hEx18_Ac24 
                 24 
                 CAG CUU CUG AGC GAG UAA UCC AGC 
                 989 
               
               
                   
                   
                   
                 UGU GAA 
                   
               
               
                   
               
               
                 19 
                 hEx19_Ac35 
                 35 
                 GCC UGA GCU GAU CUG CUG GCA UCU 
                 990 
               
               
                   
                   
                   
                 UGC AGU U 
                   
               
               
                   
               
               
                 19 
                 hEx19_Ac39 
                 39 
                 UCU GCU GGC AUC UUG C 
                 991 
               
               
                   
               
               
                 20 
                 hEx20_Ac23 
                 23 
                 GUU CAG UUG UUC UGA GGC UUG UUU 
                 992 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 20 
                 mEx20_Ac23 
                 23 
                 GUU CAG UUG UUC UGA AGC UUG UCU 
                 993 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 20 
                 hEx20_Ac44 
                 44 
                 CUG GCA GAA UUC GAU CCA CCG GCU 
                 994 
               
               
                   
                   
                   
                 GUU C 
                   
               
               
                   
               
               
                 20 
                 mEx20_Ac44 
                 44 
                 UUG GCA GAA UUC UGU CCA CCG GCU 
                 995 
               
               
                   
                   
                   
                 GUU C 
                   
               
               
                   
               
               
                 20 
                 hEx20_Ac140 
                 140 
                 AGU AGU UGU CAU CUG CUC CAA UUG 
                 996 
               
               
                   
                   
                   
                 U 
                   
               
               
                   
               
               
                 20 
                 mEx20_Ac140 
                 140 
                 AGU AGU UGU CAU CUG UUC CAA UUG 
                 997 
               
               
                   
                   
                   
                 U 
                   
               
               
                   
               
               
                 20 
                 hEx20_Ac147 
                 147 
                 CAG CAG UAG UUG UCA UCU GCU C 
                 998 
               
               
                   
               
               
                 20 
                 mEx20_Ac147 
                 147 
                 CGG CAG UAG UUG UCA UCU GUU C 
                 999 
               
               
                   
               
               
                 21 
                 hEx21_Ac85 
                 85 
                 CUG CAU CCA GGA ACA UGG GUC C 
                 1000 
               
               
                   
               
               
                 21 
                 mEx21_Ac85 
                 85 
                 CUG CAU CCA GAA ACA UUG GCC C 
                 1001 
               
               
                   
               
               
                 21 
                 hEx21_Ac86 
                 86 
                 GUC UGC AUC CAG GAA CAU GGG UC 
                 1002 
               
               
                   
               
               
                 22 
                 mEx22_Ac8 
                 8 
                 AUG UCC ACA GAC CUG UAA UU 
                 1003 
               
               
                   
               
               
                 22 
                 hEx22_Ac8 
                 8 
                 AUA UUC ACA GAC CUG CAA UU 
                 1004 
               
               
                   
               
               
                 22 
                 hEx22_Ac125 
                 125 
                 CUG CAA UUC CCC GAG UCU CUG C 
                 1005 
               
               
                   
               
               
                 22 
                 mEx22_Ac125 
                 125 
                 CUG UAA UUU CCC GAG UCU CUC C 
                 1006 
               
               
                   
               
               
                 23 
                 mEx23_Ac7 
                 7 
                 GGC CAA ACC UCG GCU UAC CUG AAA 
                 1007 
               
               
                   
                   
                   
                 U 
                   
               
               
                   
               
               
                 23 
                 hEx23_Ac7 
                 7 
                 AGU AAA AUC UUG AAU UAC CUG AAU 
                 1008 
               
               
                   
                   
                   
                 U 
                   
               
               
                   
               
               
                 23 
                 hEx23_Ac69 
                 69 
                 CGG CUA AUU UCA GAG GGC GCU UUC 
                 1009 
               
               
                   
                   
                   
                 UUC GAC 
                   
               
               
                   
               
               
                 23 
                 mEx23_Ac69 
                 69 
                 UGG CAU AUU UCU GAA GGU GCU UUC 
                 1010 
               
               
                   
                   
                   
                 UUG GCC 
                   
               
               
                   
               
               
                 24 
                 mEx24_Ac16 
                 16 
                 CAA CUU CAG CCA UCC AUU UCU GUA 
                 1011 
               
               
                   
                   
                   
                 A 
                   
               
               
                   
               
               
                 24 
                 hEx24_Ac16 
                 16 
                 CAA CUU CAG CCA UCC AUU UCU UCA 
                 1012 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 24 
                 hEx24_Ac51 
                 51 
                 CAA GGG CAG GCC AUU CCU CCU UC 
                 1013 
               
               
                   
               
               
                 24 
                 mEx24_Ac51 
                 51 
                 CCA GGG CAG GCC AUU CCU CUU UC 
                 1014 
               
               
                   
               
               
                 24 
                 mEx24_Ac78 
                 78 
                 GAG CUG UUU UUU CAG GAU UUC AGC 
                 1015 
               
               
                   
                   
                   
                 A 
                   
               
               
                   
               
               
                 24 
                 hEx24_Ac78 
                 78 
                 CAG CUG CUU UUU UAG AAU UUC UGA 
                 1016 
               
               
                   
                   
                   
                 A 
                   
               
               
                   
               
               
                 25 
                 hEx25_Ac95 
                 95 
                 UUG AGU UCU GUC UCA AGU CUC GAA 
                 1017 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 25 
                 mEx25_Ac95 
                 95 
                 CUA AGU UCU GUC UCC AGU CUG GAU 
                 1018 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 26 
                 hEx26_Ac−7 
                 −7 
                 CCU CCU UUC UGG CAU AGA CCU UCC 
                 1019 
               
               
                   
                   
                   
                 AC 
                   
               
               
                   
               
               
                 27 
                 hEx27_Ac82 
                 82 
                 UUA AGG CCU CUU GUG CUA CAG GUG 
                 1020 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 28 
                 hEx28_Ac99 
                 99 
                 CAG AGA UUU CCU CAG CUC CGC CAG 
                 1021 
               
               
                   
                   
                   
                 GA 
                   
               
               
                   
               
               
                 29 
                 hEx29_Ac15 
                 15 
                 UAU CCU CUG AAU GUC GCA UC 
                 1022 
               
               
                   
               
               
                 29 
                 hEx29_Ac18 
                 18 
                 GGU UAU CCU CUG AAU GUC GC 
                 1023 
               
               
                   
               
               
                 29 
                 hEx29_Ac45 
                 45 
                 UCU GUG CCA AUA UGC GAA UC 
                 1024 
               
               
                   
               
               
                 29 
                 hEx29_Ac57 
                 57 
                 UCC GCC AUC UGU UAG GGU CUG UGC 
                 1025 
               
               
                   
                   
                   
                 C 
                   
               
               
                   
               
               
                 29 
                 hEx29_Ac59 
                 59 
                 CCA UCU GUU AGG GUC UGU G 
                 1026 
               
               
                   
               
               
                 29 
                 hEx29_Ac105 
                 105 
                 UUA AAU GUC UCA AGU UCC 
                 1027 
               
               
                   
               
               
                 29 
                 hEx29_Ac127 
                 127 
                 GUA GUU CCC UCC AAC G 
                 1028 
               
               
                   
               
               
                 29 
                 hEx29_Ac131 
                 131 
                 CAU GUA GUU CCC UCC 
                 1029 
               
               
                   
               
               
                 30 
                 hEx30_Ac25 
                 25 
                 UCC UGG GCA GAC UGG AUG CUC UGU 
                 1030 
               
               
                   
                   
                   
                 UC 
                   
               
               
                   
               
               
                 31 
                 hEx31_Ac3 
                 3 
                 UAG UUU CUG AAA UAA CAU AUA CCU 
                 1031 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 32 
                 hEx32_Ac44 
                 44 
                 CUU GUA GAC GCU GCU CAA AAU UGG 
                 1032 
               
               
                   
                   
                   
                 CUG GUU 
                   
               
               
                   
               
               
                 33 
                 hEx33_Ac64 
                 64 
                 CCG UCU GCU UUU UCU GUA CAA UCU 
                 1033 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 34 
                 hEx34_Ac46 
                 46 
                 CAU UCA UUU CCU UUC GCA UCU UAC 
                 1034 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 34 
                 hEx34_Ac95 
                 95 
                 AUC UCU UUG UCA AUU CCA UAU CUG 
                 1035 
               
               
                   
                   
                   
                 UA 
                   
               
               
                   
               
               
                 35 
                 hEx35_Ac24 
                 24 
                 UCU GUG AUA CUC UUC AGG UGC ACC 
                 1036 
               
               
                   
                   
                   
                 UUC UGU 
                   
               
               
                   
               
               
                 36 
                 hEx36_Ac22 
                 22 
                 UGU GAU GUG GUC CAC AUU CUG GUC 
                 1037 
               
               
                   
                   
                   
                 AAA AGU 
                   
               
               
                   
               
               
                 37 
                 hEx37_Ac134 
                 134 
                 UUC UGU GUG AAA UGG CUG CAA AUC 
                 1038 
               
               
                   
               
               
                 38 
                 hEx38_Ac88 
                 88 
                 UGA AGU CUU CCU CUU UCA GAU UCA 
                 1039 
               
               
                   
                   
                   
                 C 
                   
               
               
                   
               
               
                 39 
                 hEx39_Ac62 
                 62 
                 UUU CCU CUC GCU UUC UCU CAU CUG 
                 1040 
               
               
                   
                   
                   
                 UGA UUC 
                   
               
               
                   
               
               
                 40 
                 hEx40_Ac−5 
                 -5 
                 CUU UGA GAC CUC AAA UCC UGU U 
                 1041 
               
               
                   
               
               
                 40 
                 hEx40_Ac13 
                 13 
                 GAG CCU UUU UUC UUC UUU G 
                 1042 
               
               
                   
               
               
                 40 
                 hEx40_Ac127 
                 127 
                 UCC UUU CAU CUC UGG GCU C 
                 1043 
               
               
                   
               
               
                 41 
                 hEx41_Ac44 
                 44 
                 CAA GCC CUC AGC UUG CCU ACG CAC 
                 1044 
               
               
                   
                   
                   
                 UG 
                   
               
               
                   
               
               
                 41 
                 hEx41_Ac18 
                 18 
                 CUC CUC UUU CUU CUU CUG C 
                 1045 
               
               
                   
               
               
                 41 
                 hEx41_Ac145 
                 145 
                 CUU CGA AAC UGA GCA AAU UU 
                 1046 
               
               
                   
               
               
                 42 
                 hEx42_Ac4 
                 4 
                 AUC GUU UCU UCA CGG ACA GUG UGC 
                 1047 
               
               
                   
                   
                   
                 UGG 
                   
               
               
                   
               
               
                 42 
                 hEx42_Ac90 
                 90 
                 CUU GUG AGA CAU GAG UG 
                 1048 
               
               
                   
               
               
                 42 
                 hEx42_Ac175 
                 175 
                 CAG AGA CUC CUC UUG CUU 
                 1049 
               
               
                   
               
               
                 43 
                 hEx43_Ac52 
                 52 
                 UGC UGC UGU CUU CUU GCU 
                 1050 
               
               
                   
               
               
                 43 
                 hEx43_Ac90 
                 90 
                 CUG UAG CUU CAC CCU UUC C 
                 1051 
               
               
                   
               
               
                 43 
                 hEx43_Ac101 
                 101 
                 GGA GAG AGC UUC CUG UAG CU 
                 1052 
               
               
                   
               
               
                 43 
                 hEx43_Ac132 
                 132 
                 UGU UAA CUU UUU CCC AUU GG 
                 1053 
               
               
                   
               
               
                 43 
                 hEx43_Ac134 
                 134 
                 UUG UUA ACU UUU UCC AUU 
                 1054 
               
               
                   
               
               
                 43 
                 hEx43_Ac137 
                 137 
                 CAU UUU GUU AAC UUU UUC CC 
                 1055 
               
               
                   
               
               
                 44 
                 hEx44_Ac0 
                 0 
                 CGC CAT TTC TCA ACA GAT CTG TCA 
                 1056 
               
               
                   
                   
                   
                 AAT CGC 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac1 
                 1 
                 CCG CCA TTT CTC AAC AGA TCTGTC 
                 1057 
               
               
                   
                   
                   
                 AAA TCG 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac2 
                 2 
                 GCC GCC ATT TCT CAA CAG ATC TGT 
                 1058 
               
               
                   
                   
                   
                 CAA ATC 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac3 
                 3 
                 AGC CGC CAT TTC TCA ACA GAT CTG 
                 1059 
               
               
                   
                   
                   
                 TCA AAT 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac4 
                 4 
                 AAG CCG CCA TTT CTC AAC AGA TCT 
                 1060 
               
               
                   
                   
                   
                 GTC AAA 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac5 
                 5 
                 AAA GCC GCC ATT TCT CAA CAG ATC 
                 1061 
               
               
                   
                   
                   
                 TGT CAA 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac6 
                 6 
                 AAA AGC CGC CAT TTC TCA ACA GAT 
                 1062 
               
               
                   
                   
                   
                 CTG TCA 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac7 
                 7 
                 AAA ACG CCG CCA TTT CTC AAC AGA 
                 1063 
               
               
                   
                   
                   
                 TCT GTC 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac8 
                 8 
                 GAA AAC GCC GCC ATT TCT CAA CAG 
                 1064 
               
               
                   
                   
                   
                 ATC TGT 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac9 
                 9 
                 TGA AAA CGC CGC CAT TTC TCA ACA 
                 1065 
               
               
                   
                   
                   
                 GAT CTG 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac10 
                 10 
                 ATG AAA ACG CCG CCA TTT CTC AAC 
                 1066 
               
               
                   
                   
                   
                 AGA TCT 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac14 
                 14 
                 CAT AAT GAA AAC GCC GCC ATT TCT 
                 1067 
               
               
                   
                   
                   
                 CAA CAG 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac15 
                 15 
                 CGC CGC CAU UUC UCA ACA G 
                 1068 
               
               
                   
               
               
                 44 
                 hEx44_Ac18 
                 18 
                 ATA TCA TAA TGA AAA CGC CGC CAT 
                 1069 
               
               
                   
                   
                   
                 TTC TCA 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac19 
                 19 
                 TAT ATC ATA ATG AAA ACG CCG CCA 
                 1070 
               
               
                   
                   
                   
                 TTT CTC 
                   
               
               
                   
               
               
                 44 
                 hEx44_54 
                 54 
                 TGT TCA GCT TCT GTT AGC CAC TGA 
                 1071 
               
               
                   
                   
                   
                 TTA AAT 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac56 
                 56 
                 ACT GTT CAG CTT CTG TTA GCC ACT 
                 1072 
               
               
                   
                   
                   
                 GAT TAA 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac59 
                 59 
                 GAA ACT GTT CAG CTT CTG TTA GCC 
                 1073 
               
               
                   
                   
                   
                 ACT GAT 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac61 
                 61 
                 UGU UCA GCU UCU GUU AGC CAC UGA 
                 1074 
               
               
                   
               
               
                 44 
                 hEx44_Ac69 
                 69 
                 GTC TTT CTG AGA AAC TGT TCA GCT 
                 1075 
               
               
                   
                   
                   
                 TCT GTT 
                   
               
               
                   
               
               
                 44 
                 hEx44_Ac87 
                 87 
                 UUU GUA UUU AGC AUG UUC CC 
                 1076 
               
               
                   
               
               
                 45 
                 hEx45_Ac−6 
                 −6 
                 CCA AUG CCA UCC UGG AGU UCC UGU 
                 1077 
               
               
                   
                   
                   
                 AA 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac0 
                 0 
                 TTG CCG CTG CCC AAT GCC ATC CTG 
                 1078 
               
               
                   
                   
                   
                 GAG TTC 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac1 
                 1 
                 TTT GCC GCT GCC CAA TGC CAT CCT 
                 1079 
               
               
                   
                   
                   
                 GGA GTT 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac2 
                 2 
                 GTT TGC CGC TGC CCA ATG CCA TCC 
                 1080 
               
               
                   
                   
                   
                 TGG AGT 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac3 
                 3 
                 AGT TTG CCG CTG CCC AAT GCC ATC 
                 1081 
               
               
                   
                   
                   
                 CTG GAG 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac4 
                 4 
                 CAG TTT GCC GCT GCC CAA TGC CAT 
                 1082 
               
               
                   
                   
                   
                 CCT GGA 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac6 
                 6 
                 GCC CAA UGC CAU CCU GG 
                 1083 
               
               
                   
               
               
                 45 
                 hEx45_Ac7 
                 7 
                 CAA CAG TTT GCC GCT GCC CAA TGC 
                 1084 
               
               
                   
                   
                   
                 CAT CCT 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac8 
                 8 
                 ACA ACA GTT TGC CGC TGC CCA ATG 
                 1085 
               
               
                   
                   
                   
                 CCA TCC 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac9 
                 9 
                 GAC AAC AGT TTG CCG CTG CCC AAT 
                 1086 
               
               
                   
                   
                   
                 GCC ATC 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac10 
                 10 
                 TGA CAA CAG TTT GCC GCT GCC CAA 
                 1087 
               
               
                   
                   
                   
                 TGC CAT 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac11 
                 11 
                 CTG ACA ACA GTT TGC CGC TGC CCA 
                 1088 
               
               
                   
                   
                   
                 ATG CCA 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac12 
                 12 
                 TCT GAC AAC AGT TTG CCG CTG CCC 
                 1089 
               
               
                   
                   
                   
                 AAT GCC 
                   
               
               
                   
               
               
                 45 
                 hEx45_Ac58 
                 58 
                 GCU GAA UUA UUU CUU CCC C 
                 1090 
               
               
                   
               
               
                 45 
                 hEx45_Ac75 
                 75 
                 UCU GUU UUU GAG GAU UGC 
                 1091 
               
               
                   
               
               
                 45 
                 hEx45_Ac122 
                 122 
                 CCA CCG CAG AUU CAG GC 
                 1092 
               
               
                   
               
               
                 45 
                 hEx45_Ac137 
                 137 
                 UUU GCA GAC CUC CUG CC 
                 1093 
               
               
                   
               
               
                 45 
                 hEx45_Ac154 
                 154 
                 UUU UUC UGU CUG ACA GCU G 
                 1094 
               
               
                   
               
               
                 46 
                 hEx46_Ac14 
                 14 
                 CUG ACA AGA UAU UCU U 
                 1095 
               
               
                   
               
               
                 46 
                 hEx46_Ac15 
                 15 
                 GAA AUU CUG ACA AGA UAU UCU 
                 1096 
               
               
                   
               
               
                 46 
                 hEx46_Ac45 
                 45 
                 CTT CCT CCA ACC ATA AAA CAA ATT 
                 1097 
               
               
                   
                   
                   
                 CAT TTA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac46 
                 46 
                 GCT TCC TCC AAC CAT AAA ACA AAT 
                 1098 
               
               
                   
                   
                   
                 TCA TTT 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac47 
                 4 
                 TGC TTC CTC CAA CCA TAA AAC AAA 
                 1099 
               
               
                   
                   
                   
                 TTC ATT 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac47 
                 47 
                 UAA AAC AAA UUC AUU 
                 1100 
               
               
                   
               
               
                 46 
                 hEx46_Ac48 
                 48 
                 CTG CTT CCT CCA ACC ATA AAA CAA 
                 1101 
               
               
                   
                   
                   
                 ATT CAT 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac49 
                 49 
                 TCT GCT TCC TCC AAC CAT AAA ACA 
                 1102 
               
               
                   
                   
                   
                 AAT TCA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac50 
                 50 
                 ATC TGC TTC CTC CAA CCA TAA AAC 
                 1103 
               
               
                   
                   
                   
                 AAA TTC 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac51 
                 51 
                 TAT CTG CTT CCT CCA ACC ATA AAA 
                 1104 
               
               
                   
                   
                   
                 CAA ATT 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac52 
                 52 
                 TTA TCT GCT TCC TCC AAC CAT AAA 
                 1105 
               
               
                   
                   
                   
                 ACA AAT 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac53 
                 53 
                 GTT ATC TGC TTC CTC CAA CCA TAA 
                 1106 
               
               
                   
                   
                   
                 AAC AAA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac54 
                 54 
                 TGT TAT CTG CTT CCT CCA ACC ATA 
                 1107 
               
               
                   
                   
                   
                 AAA CAA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac55 
                 55 
                 ATG TTA TCT GCT TCC TCC AAC CAT 
                 1108 
               
               
                   
                   
                   
                 AAA ACA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac56 
                 56 
                 AAT GTT ATC TGC TTC CTC CAA CCA 
                 1109 
               
               
                   
                   
                   
                 TAA AAC 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac57 
                 57 
                 CAA TGT TAT CTG CTT CCT CCA ACC 
                 1110 
               
               
                   
                   
                   
                 ATA AAA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac58 
                 58 
                 GCA ATG TTA TCT GCT TCC TCC AAC 
                 1111 
               
               
                   
                   
                   
                 CAT AAA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac59 
                 59 
                 AGC AAT GTT ATC TGC TTC CTC CAA 
                 1112 
               
               
                   
                   
                   
                 CCA TAA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac60 
                 60 
                 TAG CAA TGT TAT CTG CTT CCT CCA 
                 1113 
               
               
                   
                   
                   
                 ACC ATA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac61 
                 61 
                 CTA GCA ATG TTA TCT GCT TCC TCC 
                 1114 
               
               
                   
                   
                   
                 AAC CAT 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac62 
                 62 
                 ACT AGC AAT GTT ATC TGC TTC CTC 
                 1115 
               
               
                   
                   
                   
                 CAA CCA 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac63 
                 63 
                 GUU AUC UGC UUC CUC CAA CC 
                 1116 
               
               
                   
               
               
                 46 
                 hEx46_Ac88 
                 88 
                 AGG UUC AAG UGG GAU ACU A 
                 1117 
               
               
                   
               
               
                 46 
                 hEx46_Ac90 
                 90 
                 UCC AGG UUC AAG UGG GAU AC 
                 1118 
               
               
                   
               
               
                 46 
                 hEx46_Ac96 
                 96 
                 UUC CAG GUU CAA GUG 
                 1119 
               
               
                   
               
               
                 46 
                 hEx46_Ac107 
                 107 
                 CAA GCU UUU CUU UUA GUU GCU GCU 
                 1120 
               
               
                   
                   
                   
                 CUU UUC C 
                   
               
               
                   
               
               
                 46 
                 hEx46_Ac11 
                 111 
                 UUA GUU GCU GCU CUU 
                 1121 
               
               
                   
               
               
                 46 
                 hEx46_Ac115 
                 115 
                 GCU UUU CUU UUA GUU GCU GC 
                 1122 
               
               
                   
               
               
                 46 
                 hEx46_Ac122 
                 122 
                 UCA AGC UUU UCU UUU AG 
                 1123 
               
               
                   
               
               
                 47 
                 hEx47_Ac−6 
                 −6 
                 CAG GGG CAA CUC UUC CAC CAG UAA 
                 1124 
               
               
                   
                   
                   
                 CUG AAA 
                   
               
               
                   
               
               
                 47 
                 hEx47_Ac39 
                 39 
                 UCC AGU UUC AUU UAA UUG UUU G 
                 1125 
               
               
                   
               
               
                 47 
                 hEx47_Ac63 
                 63 
                 AGC ACU UAC AAG CAC GGG U 
                 1126 
               
               
                   
               
               
                 47 
                 hEx47_Ac87 
                 87 
                 UCU UGC UCU UCU GGG CUU 
                 1127 
               
               
                   
               
               
                 47 
                 hEx47_Ac94 
                 94 
                 UUC AAG UUU AUC UUG CUC UUC 
                 1128 
               
               
                   
               
               
                 47 
                 hEx47_Ac101 
                 101 
                 CUU GAG CUU AUU UUC AAG UUU 
                 1129 
               
               
                   
               
               
                 47 
                 hEx47_Ac103 
                 103 
                 CUG CUU GAG CUU AUU UUC AAG UU 
                 1130 
               
               
                   
               
               
                 48 
                 hEx48_Ac−7 
                 −7 
                 UUC UCA GGU AAA GCU CUG GAA ACC 
                 1131 
               
               
                   
                   
                   
                 UGA AAG 
                   
               
               
                   
               
               
                 48 
                 hEx48_Ac2 
                 2 
                 CUU CAA GCU UUU UUU CAA GCU 
                 1132 
               
               
                   
               
               
                 48 
                 hEx48_Ac19 
                 19 
                 UUU CUC CUU GUU UCU C 
                 1133 
               
               
                   
               
               
                 48 
                 hEx48_Ac23 
                 23 
                 GCU UCA AUU UCU CCU UGU U 
                 1134 
               
               
                   
               
               
                 48 
                 hEx48_Ac32 
                 32 
                 UUU AUU UGA GCU UCA AUU U 
                 1135 
               
               
                   
               
               
                 48 
                 hEx48_Ac37 
                 37 
                 GGU CUU UUA UUU GAG CUU C 
                 1136 
               
               
                   
               
               
                 48 
                 hEx48_Ac48 
                 48 
                 GCU GCC CAA GGU CUU UU 
                 1137 
               
               
                   
               
               
                 48 
                 hEx48_Ac71 
                 71 
                 CUU CAA GGU CUU CAA GCU UUU 
                 1138 
               
               
                   
               
               
                 48 
                 hEx48_Ac79 
                 79 
                 UAA CUG CUC UUC AAG GUC UUC 
                 1139 
               
               
                   
               
               
                 48 
                 hEx48_Ac133 
                 133 
                 UUA UAA AUU UCC AAC UGA UUC 
                 1140 
               
               
                   
               
               
                 49 
                 hEx49_Ac−11 
                 −11 
                 CUG CUA UUU CAG UUU CCU GGG GAA 
                 1141 
               
               
                   
                   
                   
                 AAG 
                   
               
               
                   
               
               
                 49 
                 hEx49_Ac25 
                 25 
                 CUU CCA CAU CCG GUU GUU U 
                 1142 
               
               
                   
               
               
                 49 
                 hEx49_Ac60 
                 60 
                 GUG GCU GGU UUU UCC UUG U 
                 1143 
               
               
                   
               
               
                 50 
                 hEx50_Ac2 
                 2 
                 CCA CUC AGA GCU CAG AUC UUC UAA 
                 1144 
               
               
                   
                   
                   
                 CUU CC 
                   
               
               
                   
               
               
                 50 
                 hEx50_Ac11 
                 11 
                 CUC AGA GCU CAG AUC UU 
                 1145 
               
               
                   
               
               
                 50 
                 hEx50_Ac36 
                 36 
                 GGC UGC UUU GCC CUC 
                 1146 
               
               
                   
               
               
                 51 
                 hEx51_Ac0 
                 0 
                 GTG TCA CCA GAG TAA CAG TCT GAG 
                 1147 
               
               
                   
                   
                   
                 TAG GAG 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac5 
                 5 
                 AGG TTG TGT CAC CAG AGT AAC AGT 
                 1148 
               
               
                   
                   
                   
                 CTG AGT 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac9 
                 9 
                 CCA CAG GTT GTG TCA CCA GAG TAA 
                 1149 
               
               
                   
                   
                   
                 CAG TCT 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac26 
                 26 
                 GGC AGT TTC CTT AGT AAC CAC AGG 
                 1150 
               
               
                   
                   
                   
                 TTG TGT 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac30 
                 30 
                 AGA TGG CAG TTT CCT TAG TAA CCA 
                 1151 
               
               
                   
                   
                   
                 CAG GTT 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac48 
                 48 
                 ATG GCA TTT CTA GTT TGG AGA TGG 
                 1152 
               
               
                   
                   
                   
                 CAG TTT 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac65 
                 65 
                 CTC CAA CAT CAA GGA AGA TGG CAT 
                 1153 
               
               
                   
                   
                   
                 TTC TAG 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac66 
                 66 
                 ACA UCA AGG AAG AUG GCA UUU CUA 
                 1154 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac67 
                 67 
                 TCA AGG AAG ATG GCA TTT CT 
                 1155 
               
               
                   
               
               
                 51 
                 hEx51_Ac68 
                 68 
                 UCA AGG AAG AUG GCA UUU CU 
                 1156 
               
               
                   
               
               
                 51 
                 hEx51_Ac132 
                 132 
                 GAA AGC CAG UCG GUA AGU UC 
                 1157 
               
               
                   
               
               
                 51 
                 hEx51_Ac141 
                 141 
                 TTA TAA CTT GAT CAA GCA GAG AAA 
                 1158 
               
               
                   
                   
                   
                 GCC AGT 
                   
               
               
                   
               
               
                 51 
                 hEx51_Ac160 
                 160 
                 CCU CUG UGA UUU UAU AAC UUG AU 
                 1159 
               
               
                   
               
               
                 51 
                 hEx51_Ac181 
                 181 
                 CAC CCA CCA UCA CCC 
                 1160 
               
               
                   
               
               
                 51 
                 hEx51_Ac191 
                 191 
                 UGA UAU CCU CAA GGU CAC CC 
                 1161 
               
               
                   
               
               
                 51 
                 hEx51_Ac207 
                 207 
                 ATA CCT TCT GCT TGA TGA TCA TCT 
                 1162 
               
               
                   
                   
                   
                 CGT TGA 
                   
               
               
                   
               
               
                 52 
                 hEx52_Ac12 
                 12 
                 UCC AAC UGG GGA CGC CUC UGU UCC 
                 1163 
               
               
                   
                   
                   
                 AAA UCC 
                   
               
               
                   
               
               
                 52 
                 mEx52_Ac12 
                 12 
                 UCC AAU UGG GGG CGU CUC UGU UCC 
                 1164 
               
               
                   
                   
                   
                 AAA UCU 
                   
               
               
                   
               
               
                 52 
                 mEx52_Ac17 
                 17 
                 UCC AAU UGG GGG CGU CUC UGU UCC 
                 1165 
               
               
                   
                   
                   
                 A 
                   
               
               
                   
               
               
                 52 
                 hEx52_Ac17 
                 17 
                 UCC AAC UGG GGA CGC CUC UGU UCC 
                 1166 
               
               
                   
                   
                   
                 A 
                   
               
               
                   
               
               
                 52 
                 hEx52_Ac18 
                 18 
                 UUC CAA CUG GGG ACG CCU CUG UUC 
                 1167 
               
               
                   
                   
                   
                 C 
                   
               
               
                   
               
               
                 52 
                 hEx52_Ac24 
                 24 
                 GGT AAT GAG TTC TTC CAA CTG GGG 
                 1168 
               
               
                   
                   
                   
                 ACG CCT 
                   
               
               
                   
               
               
                 52 
                 mEx52_Ac42 
                 42 
                 UUC AAA UUC UGG GCA GCA GUA AUG 
                 1169 
               
               
                   
                   
                   
                 AGU UCU 
                   
               
               
                   
               
               
                 52 
                 hEx52_Ac42 
                 42 
                 UUC AAA UUU UGG GCA GCG GUA AUG 
                 1170 
               
               
                   
                   
                   
                 AGU UCU 
                   
               
               
                   
               
               
                 52 
                 hEx52_Ac69 
                 69 
                 UUG CUG GUC UUG UUU UUC 
                 1171 
               
               
                   
               
               
                 52 
                 hEx52_Ac97 
                 97 
                 CCG UAA UGA UUG UUC U 
                 1172 
               
               
                   
               
               
                 53 
                 hEx53_Ac1 
                 1 
                 ACT TCA TCC CAC TGA TTC TGA ATT 
                 1173 
               
               
                   
                   
                   
                 CTT TCA 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac2 
                 2 
                 TAC TTC ATC CCA CTG ATT CTG AAT 
                 1174 
               
               
                   
                   
                   
                 TCT TTC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac3 
                 3 
                 GTA CTT CAT CCC ACT GAT TCT GAA 
                 1175 
               
               
                   
                   
                   
                 TTC TTT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac4 
                 4 
                 TGT ACT TCA TCC CAC TGA TTC TGA 
                 1176 
               
               
                   
                   
                   
                 ATT CTT 
                   
               
               
                   
               
               
                 53 
                 mEx53_Ac5 
                 5 
                 UUU UAA AGA UAU GCU UGA CAC UAA 
                 1177 
               
               
                   
                   
                   
                 CCU UGG 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac5 
                 5 
                 UUA AAA AGG UAU CUU UGA UAC UAA 
                 1178 
               
               
                   
                   
                   
                 CCU UGG 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac5 
                 5 
                 TTG TAC TTC ATC CCA CTG ATT CTG 
                 1179 
               
               
                   
                   
                   
                 AAT TCT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac6 
                 6 
                 CTT GTA CTT CAT CCC ACT GAT TCT 
                 1180 
               
               
                   
                   
                   
                 GAA TTC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac7 
                 7 
                 TCT TGT ACT TCA TCC CAC TGA TTC 
                 1181 
               
               
                   
                   
                   
                 TGA ATT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac8 
                 8 
                 TTC TTG TAC TTC ATC CCA CTG ATT 
                 1182 
               
               
                   
                   
                   
                 CTG AAT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac9 
                 9 
                 GTT CTT GTA CTT CAT CCC ACT GAT 
                 1183 
               
               
                   
                   
                   
                 TCT GAA 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac10 
                 10 
                 TGT TCT TGT ACT TCA TCC CAC TGA 
                 1184 
               
               
                   
                   
                   
                 TTC TGA 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac11 
                 11 
                 GTG TTC TTG TAC TTC ATC CCA CTG 
                 1185 
               
               
                   
                   
                   
                 ATT CTG 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac12 
                 12 
                 GGT GTT CTT GTA CTT CAT CCC ACT 
                 1186 
               
               
                   
                   
                   
                 GAT TCT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac13 
                 13 
                 AGG TGT TCT TGT ACT TCA TCC CAC 
                 1187 
               
               
                   
                   
                   
                 TGA TTC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac14 
                 14 
                 AAG GTG TTC TTG TAC TTC ATC CCA 
                 1188 
               
               
                   
                   
                   
                 CTG ATT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac15 
                 15 
                 GAA GGT GTT CTT GTA CTT CAT CCC 
                 1189 
               
               
                   
                   
                   
                 ACT GAT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac16 
                 16 
                 TGA AGG TGT TCT TGT ACT TCA TCC 
                 1190 
               
               
                   
                   
                   
                 CAC TGA 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac17 
                 17 
                 CTG AAG GTG TTC TTG TAC TTC ATC 
                 1191 
               
               
                   
                   
                   
                 CCA CTG 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac18 
                 18 
                 TCT GAA GGT GTT CTT GTA CTT CAT 
                 1192 
               
               
                   
                   
                   
                 CCC ACT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac19 
                 19 
                 TTC TGA AGG TGT TCT TGT ACT TCA 
                 1193 
               
               
                   
                   
                   
                 TCC CAC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac20 
                 20 
                 GTT CTG AAG GTG TTC TTG TAC TTC 
                 1194 
               
               
                   
                   
                   
                 ATC CCA 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac21 
                 21 
                 GGT TCT GAA GGT GTT CTT GTA CTT 
                 1195 
               
               
                   
                   
                   
                 CAT CCC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac22 
                 22 
                 CGG TTC TGA AGG TGT TCT TGT ACT 
                 1196 
               
               
                   
                   
                   
                 TCA TCC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac23 
                 23 
                 CCG GTT CTG AAG GTG TTC TTG TAC 
                 1197 
               
               
                   
                   
                   
                 TTC ATC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac24 
                 24 
                 TCC GGT TCT GAA GGT GTT CTT GTA 
                 1198 
               
               
                   
                   
                   
                 CTT CAT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac25 
                 25 
                 CTC CGG TTC TGA AGG TGT TCT TGT 
                 1199 
               
               
                   
                   
                   
                 ACT TCA 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac26 
                 26 
                 CCT CCG GTT CTG AAG GTG TTC TTG 
                 1200 
               
               
                   
                   
                   
                 TAC TTC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac27 
                 27 
                 GCC TCC GGT TCT GAA GGT GTT CTT 
                 1201 
               
               
                   
                   
                   
                 GTA CTT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac28 
                 28 
                 TGC CTC CGG TTC TGA AGG TGT TCT 
                 1202 
               
               
                   
                   
                   
                 TGT ACT 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac20 
                 29 
                 TTG CCT CCG GTT CTG AAG GTG TTC 
                 1203 
               
               
                   
                   
                   
                 TTG TAC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac30 
                 30 
                 GTT GCC TCC GGT TCT GAA GGT GTT 
                 1204 
               
               
                   
                   
                   
                 CTT GTA 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac39 
                 39 
                 CAU UCA ACU GUU GCC UCC GGU UCU 
                 1205 
               
               
                   
                   
                   
                 GAA GGU G 
                   
               
               
                   
               
               
                 53 
                 mEx53_Ac39 
                 39 
                 CAU UCA ACU GUU GUC UCC UGU UCU 
                 1206 
               
               
                   
                   
                   
                 GCA GCU G 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac45 
                 45 
                 CUG UUG CCU CCG GUU CUG 
                 1207 
               
               
                   
               
               
                 53 
                 hEx53_Ac69 
                 69 
                 CAG CCA UUG UGU UGA AUC CUU UAA 
                 1208 
               
               
                   
                   
                   
                 CAU UUC 
                   
               
               
                   
               
               
                 53 
                 hEx53_Ac128 
                 128 
                 UUG GCU CUG GCC UGU CCU 
                 1209 
               
               
                   
               
               
                 53 
                 mEx53_Ac151 
                 151 
                 CUA CUG UGU GAG GAC CUU CUU UCC 
                 1210 
               
               
                   
                   
                   
                 AUG AGU 
                   
               
               
                   
               
               
                 53 
                 mEx53_Ac176 
                 176 
                 UCU GUG AUC UUC UUU UGG AUU GCA 
                 1211 
               
               
                   
                   
                   
                 UCU ACU 
                   
               
               
                   
               
               
                 54 
                 hEx54_Ac21 
                 21 
                 UAC AUU UGU CUG CCA CUG G 
                 1212 
               
               
                   
               
               
                 54 
                 hEx54_Ac42 
                 42 
                 GAG AAG TTT CAG GGC CAA GTC ATT 
                 1213 
               
               
                   
                   
                   
                 TGC CAC 
                   
               
               
                   
               
               
                 54 
                 hEx54_Ac58 
                 58 
                 CCC GGA GAA GUU UCA GGG 
                 1214 
               
               
                   
               
               
                 54 
                 hEx54_Ac67 
                 67 
                 UCU GCA GAA UAA UCC CGG AGA AG 
                 1215 
               
               
                   
               
               
                 55 
                 hEx55_Ac0 
                 0 
                 TCT TCC AAA GCA GCC TCT CGC TCA 
                 1216 
               
               
                   
                   
                   
                 CTC ACC 
                   
               
               
                   
               
               
                 55 
                 hEx55_Ac29 
                 29 
                 UGC AGU AAU CUA UGA GUU UC 
                 1217 
               
               
                   
               
               
                 55 
                 hEx55_Ac33 
                 33 
                 CUG UUG CAG UAA UCU AUG AG 
                 1218 
               
               
                   
               
               
                 55 
                 hEx55_Ac104 
                 104 
                 UCC UGU AGG ACA TUG GCA GU 
                 1219 
               
               
                   
               
               
                 55 
                 hEx55_Ac139 
                 139 
                 GAG UCU UCU AGG AGC CUU 
                 1220 
               
               
                   
               
               
                 55 
                 hEx55_Ac141 
                 141 
                 CUU GGA GUC UUC UAG GAG CC 
                 1221 
               
               
                   
               
               
                 55 
                 hEx55_Ac167 
                 167 
                 UGC CAU UGU UUC AUC AGC UCU UU 
                 1222 
               
               
                   
               
               
                 56 
                 hEx56_Ac48 
                 48 
                 UUU UUU GGC UGU UUU CAU CC 
                 1223 
               
               
                   
               
               
                 56 
                 hEx56_Ac69 
                 69 
                 CCU UCC AGG GAU CUC AGG 
                 1224 
               
               
                   
               
               
                 56 
                 hEx56_Ac102 
                 102 
                 GUU AUC CAA ACG UCU UUG UAA CAG 
                 1225 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 56 
                 hEx56_Ac129 
                 129 
                 GUU CAC UCC ACU UGA AGU UC 
                 1226 
               
               
                   
               
               
                 57 
                 hEx57_Ac−12 
                 -12 
                 CUG GCU UCC AAA UGG GAC CUG AAA 
                 1227 
               
               
                   
                   
                   
                 AAG AAC 
                   
               
               
                   
               
               
                 57 
                 hEx57_Ac64 
                 64 
                 UUC AGC UGU AGC CAC ACC 
                 1228 
               
               
                   
               
               
                 57 
                 hEx57_Ac97 
                 97 
                 UAG GUG CCU GCC GGC UU 
                 1229 
               
               
                   
               
               
                 57 
                 hEx57_Ac118 
                 118 
                 CUG AAC UGC UGG AAA GUC GCC 
                 1230 
               
               
                   
               
               
                 58 
                 hEx58_Ac9 
                 9 
                 UUC UUU AGU UUU CAA UUC CCU C 
                 1231 
               
               
                   
               
               
                 58 
                 hEx58_Ac21 
                 21 
                 ACU CAU GAU UAC ACG UUC UUU AGU 
                 1232 
               
               
                   
                   
                   
                 U 
                   
               
               
                   
               
               
                 58 
                 hEx58_Ac86 
                 86 
                 GAG UUU CUC UAG UCC UUC C 
                 1233 
               
               
                   
               
               
                 59 
                 hEx59_Ac6 
                 6 
                 UCC UCA GGA GGC AGC UCU AAA U 
                 1234 
               
               
                   
               
               
                 59 
                 hEx59_Ac66 
                 66 
                 GAG UUU CUC UAG UCC UUC C 
                 1235 
               
               
                   
               
               
                 59 
                 hEx59_Ac134 
                 134 
                 UUG AAG UUC CUG GAG UCU U 
                 1236 
               
               
                   
               
               
                 60 
                 hEx60_Ac19 
                 19 
                 GUU CUC UUU CAG AGG CGC 
                 1237 
               
               
                   
               
               
                 60 
                 hEx60_Ac37 
                 37 
                 CUG GCG AGC AAG GUC CUU GAC GUG 
                 1238 
               
               
                   
                   
                   
                 GCU CAC 
                   
               
               
                   
               
               
                 60 
                 hEx60_Ac92 
                 92 
                 GUG CUG AGG UUA UAC GGU G 
                 1239 
               
               
                   
               
               
                 61 
                 hEx61_Ac10 
                 10 
                 GGG CUU CAU GCA GCU GCC UGA CUC 
                 1240 
               
               
                   
                   
                   
                 GGU CCU C 
                   
               
               
                   
               
               
                 61 
                 hEx61_Ac31 
                 31 
                 GUC CCU GUG GGC UUC AUG 
                 1241 
               
               
                   
               
               
                 61 
                 hEx61_Ac51 
                 51 
                 GUG CUG AGA UGC UGG ACC 
                 1242 
               
               
                   
               
               
                 62 
                 hEx62_Ac8 
                 8 
                 GAG AUG GCU CUC UCC CAG GGA CCC 
                 1243 
               
               
                   
                   
                   
                 UGG 
                   
               
               
                   
               
               
                 62 
                 hEx62_Ac15 
                 15 
                 UGG CUC UCU CCC AGG G 
                 1244 
               
               
                   
               
               
                 62 
                 hEx62_Ac37 
                 37 
                 GGG CAC UUU GUU UGG CG 
                 1245 
               
               
                   
               
               
                 63 
                 hEx63_Ac11 
                 11 
                 UGG GAU GGU CCC AGC AAG UUG UUU 
                 1246 
               
               
                   
                   
                   
                 G 
                   
               
               
                 63 
                 hEx63_Ac11 
                 11 
                 GGU CCC AGC AAG UUG UUU G 
                 1247 
               
               
                   
               
               
                 63 
                 hEx63_Ac33 
                 33 
                 GUA GAG CUC UGU CAU UUU GGG 
                 1248 
               
               
                   
               
               
                 64 
                 hEx64_Ac47 
                 47 
                 GCA AAG GGC CUU CUG CAG UCU UCG 
                 1249 
               
               
                   
                   
                   
                 GAG 
                   
               
               
                   
               
               
                 65 
                 hEx65_Ac−11 
                 −11 
                 GCU CAA GAG AUC CAC UGC AAA AAA 
                 1250 
               
               
                   
                   
                   
                 C 
                   
               
               
                   
               
               
                 65 
                 mEx65_Ac−11 
                 −11 
                 GCU CAA GAG AUC CAC UGC AAA AAA 
                 1251 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 65 
                 hEx65_Ac15 
                 15 
                 GCC AUA CGU ACG UAU CAU AAA CAU 
                 1252 
               
               
                   
                   
                   
                 UC 
                   
               
               
                   
               
               
                 65 
                 hEx65_Ac26 
                 26 
                 GUU GUG CUG GUC CAA GGC AUC ACA 
                 1253 
               
               
                   
                   
                   
                 U 
                   
               
               
                   
               
               
                 65 
                 mEx65_Ac26 
                 26 
                 GUU GUG CUG GUC CAG GGC AUC ACA 
                 1254 
               
               
                   
                   
                   
                 U 
                   
               
               
                   
               
               
                 65 
                 hEx65_Ac63 
                 63 
                 UCU GCA GGA UAU CCA UGG GCU GGU 
                 1255 
               
               
                   
                   
                   
                 C 
                   
               
               
                   
               
               
                 65 
                 hEx65_Ac63 
                 63 
                 UCU GCA GGA UAU CCA UGG GCU GGU 
                 1256 
               
               
                   
                   
                   
                 C 
                   
               
               
                   
               
               
                 66 
                 hEx66_Ac-8 
                 −8 
                 GAU CCU CCC UGU UCG UCC CCU AUU 
                 1257 
               
               
                   
                   
                   
                 AUG 
                   
               
               
                   
               
               
                 67 
                 hEx67_Ac22 
                 22 
                 GCG CUG GUC ACA AAA UCC UGU UGA 
                 1258 
               
               
                   
                   
                   
                 AC 
                   
               
               
                   
               
               
                 68 
                 hEx68_Ac22 
                 22 
                 CAU CCA GUC UAG GAA GAG GGC CGC 
                 1259 
               
               
                   
                   
                   
                 UUC 
                   
               
               
                   
               
               
                 69 
                 hEx69_Ac−6 
                 −6 
                 UGC UUU AGA CUC CUG UAC CUG AUA 
                 1260 
               
               
                   
               
               
                 70 
                 hEx70_Ac98 
                 98 
                 CCU CUA AGA CAG UCU GCA CUG GCA 
                 1261 
               
               
                   
               
               
                 71 
                 hEx71_Ac−3 
                 −3 
                 AAG UUG AUC AGA GUA ACG GGA CUG 
                 1262 
               
               
                   
               
               
                 71 
                 hEx71_Ac8 
                 8 
                 GCC AGA AGU UGA UCA GAG U 
                 1263 
               
               
                   
               
               
                 71 
                 hEx71_Ac16 
                 16 
                 UCU ACU GGC CAG AAG UUG 
                 1264 
               
               
                   
               
               
                 72 
                 hEx72_Ac2 
                 2 
                 GUG UGA AAG CUG AGG GGA CGA GGC 
                 1265 
               
               
                   
                   
                   
                 AGG 
                   
               
               
                   
               
               
                 72 
                 hEx72_Ac20 
                 20 
                 UGA GUA UCA UCG UGU GAA AG 
                 1266 
               
               
                   
               
               
                 72 
                 hEx72_Ac42 
                 42 
                 GCA UAA UGU UCA AUG CGU G 
                 1267 
               
               
                   
               
               
                 73 
                 hEx73_Ac6 
                 6 
                 GAU CCA UUG CUG UUU UCC AUU UCU 
                 1268 
               
               
                   
                   
                   
                 G 
                   
               
               
                   
               
               
                 73 
                 hEx73_Ac13 
                 13 
                 GAU CCA UUG CUG UUU UCC 
                 1269 
               
               
                   
               
               
                 73 
                 hEx73_AC31 
                 31 
                 GAG AUG CUA UCA UUU AGA UAA 
                 1270 
               
               
                   
               
               
                 74 
                 hEx74_Ac48 
                 48 
                 CGA GGC UGG CUC AGG GGG GAG UCC 
                 1271 
               
               
                   
                   
                   
                 U 
                   
               
               
                   
               
               
                 74 
                 hEx74_Ac51 
                 51 
                 CUG GCU CAG GGG GGA GU 
                 1272 
               
               
                   
               
               
                 74 
                 hEx74_Ac72 
                 72 
                 UCC CCU CUU UCC UCA CUC U 
                 1273 
               
               
                   
               
               
                 75 
                 hEx75_Ac34 
                 34 
                 GGA CAG GCC UUU AUG UUC GUG CUG 
                 1274 
               
               
                   
                   
                   
                 C 
                   
               
               
                   
               
               
                 75 
                 hEx75_Ac33 
                 33 
                 CCU MIA UGU UCG UGC UGC U 
                 1275 
               
               
                   
               
               
                 75 
                 hEx75_Ac144 
                 144 
                 GGC GGC CUU UGU GUU GAC 
                 1276 
               
               
                   
               
               
                 76 
                 hEx76_Ac53 
                 53 
                 GCU GAC UGC UGU CGG ACC UCU GUA 
                 1277 
               
               
                   
                   
                   
                 GAG 
                   
               
               
                   
               
               
                 76 
                 hEx76_Ac37 
                 37 
                 GAG AGG UAG AAG GAG AGG A 
                 1278 
               
               
                   
               
               
                 76 
                 hEx76_Ac65 
                 65 
                 AUA GGC UGA CUG CUG UCG G 
                 1279 
               
               
                   
               
               
                 77 
                 hEx77_Ac16 
                 16 
                 CUG UGC UUG UGU CCU GGG GAG GAC 
                 1280 
               
               
                   
                   
                   
                 UGA 
                   
               
               
                   
               
               
                 77 
                 hEx77_Ac20 
                 20 
                 UUG UGU CCU GGG GAG GA 
                 1281 
               
               
                   
               
               
                 77 
                 hEx77_A47 
                 47 
                 UGC UCC AUC ACC UCC UCU 
                 1282 
               
               
                   
               
               
                 78 
                 hEx78_Ac4 
                 4 
                 UCU CAU UGG CUU UCC AGG GGU AUU 
                 1283 
               
               
                   
                   
                   
                 UC 
                   
               
               
                   
               
               
                 78 
                 hEx78_Ac4 
                 4 
                 GCU UUC CAG GGG UAU UUC 
                 1284 
               
               
                   
               
               
                 78 
                 hEx78_Ac10 
                 10 
                 CAU UGG CUU UCC AGG GG 
                 1285 
               
               
                   
               
            
           
         
       
     
     Example 9. Screening of DMD Exon 44 and 45 Skipping PMOs in Transfected Primary Human Skeletal Muscle Cells 
     Primary, pre-differentiated human skeletal muscle cells (Gibco, # A11440) were plated on collagen Type 1 coated 24-well plates (Gibco, #1970788) in DMEM supplemented with 2% horse serum) and 1×ITS (Gibco, #1933286) according to the manufacturer&#39;s instructions. Cells were grown in 37° C.+5% CO 2  for 2 days to establish myotubes. These cells were then treated with defined concentrations of PMOs in water and 2 uM Endo-Porter (Gene Tools, # EP6P1-1) to facilitate PMO uptake into cells. Cell were harvested 48 hours after treatment by aspirating the culture medium and addition of 300 ul TRIZOL per well. Cells were frozen at −80° C. before RNA was prepared using Direct-zol T -96 RNA kit (Zymo Research, # R2056). Total RNA concentration was quantified spectroscopically. Between 100-200 ng total RNA was reverse transcribed using High Capacity cDNA Reverse Transcription kit (Applied Biosystems, #4368813). RT PCR reactions were incubated at 25° C. for 10 min, 37° C. for 120 min, 85° C. for 5 min, and then held at 4° C. Reactions were diluted 1:1 with water. For quantification of exon skipping by gel electrophoresis DNA fragments representing total (non-skipped+skipped) and skipped mRNAs were amplified by qPCR using Taqman Fast Advanced Master mix (Applied Biosystems, #4444558) and specific primer pairs (see Table 19). qPCR reactions were incubated at 95° C. for 20 sec, followed by 32 cycles of 95° C. for 1 sec and 60° C. for 20 sec using a QuantStudio 7 Flex (Applied Biosystems). PCR products were diluted 4:1 with TAE loading buffer and loaded onto 24-well 4% TAE gels (Embi Tec, # GG3807) containing GelGreen. PCR products were separated by electrophoresis (50 V for 2 hrs). The intensity of bands corresponding to total DMD and skipped DMD products were quantified by densiometry using ChemiDoc m XRS+(Bio-Rad). 
     Taqman qPCR primers and probes are illustrated in Table 19. 
     
       
         
           
               
               
               
             
               
                   
               
             
            
               
                 hDMD Ex44 skipped 
                 Forward: 
                 5′-CTGTGGAAAGGGTGAAG 
               
               
                   
                   
                 CTA-3′ 
               
               
                   
                 Reverse: 
                 5′-GACAAGGGAACTCCAGG 
               
               
                   
                   
                 ATG-3 
               
               
                   
                 Probe: 
                 5′-AGCTCTCTCCCAGCTTG 
               
               
                   
                   
                 ATTTCCA-3′ 
               
               
                   
               
               
                 hDMD Ex45 skipped 
                 Forward: 
                 5′-CAGTGGCTAACAGAAGC 
               
               
                   
                   
                 TGA-3′ 
               
               
                   
                 Reverse: 
                 5′-CAAATGGTATCTTAAGG 
               
               
                   
                   
                 CTAGAAGAAC-3′ 
               
               
                   
                 Probe: 
                 5′-ACACAAATTCCTGAGAA 
               
               
                   
                   
                 TTGGGAACATGC-3′ 
               
               
                   
               
            
           
         
       
     
     hDMD total Hs01049401_m1, human DMD VIC-MGB, 360 rxns (Thermo Fisher Scientific) 
     Table 20A illustrates exon skipping activity of PMOs (30mer) targeting DMD exon 45 in transfected primary human skeletal muscle cells. 
     
       
         
           
               
               
               
            
               
                   
               
               
                   
                 PMO conc 
                 % Skipping (skipped/total) 
               
            
           
           
               
               
               
               
            
               
                   
                 uM 
                 AVG 
                 STDEV 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 hEx45_Ac1 
                 10.0 
                 43.5 
                 6.4 
               
               
                   
                 3.0 
                 38.5 
                 9.2 
               
               
                   
                 1.0 
                 29.5 
                 3.5 
               
               
                 hEx45_Ac2 
                 10.0 
                 67.0 
                 14.1 
               
               
                   
                 3.0 
                 71.5 
                 14.8 
               
               
                   
                 1.0 
                 38.0 
                 7.8 
               
               
                   
                 0.1 
                 10.0 
                   
               
               
                 hEx45_Ac3 
                 10.0 
                 69.5 
                 2.1 
               
               
                   
                 3.0 
                 56.5 
                 10.6 
               
               
                   
                 1.0 
                 34.0 
                 8.5 
               
               
                 hEx45_Ac4 
                 10.0 
                 51.7 
                 10.4 
               
               
                   
                 3.0 
                 49.0 
                 1.4 
               
               
                   
                 1.0 
                 34.0 
                 5.3 
               
               
                   
                 0.1 
                 18.0 
                   
               
               
                 hEx45_Ac7 
                 10.0 
                 72.0 
                 11.4 
               
               
                   
                 3.0 
                 62.5 
                 2.1 
               
               
                   
                 1.0 
                 43.3 
                 4.9 
               
               
                   
                 0.1 
                 18.0 
                   
               
               
                 hEx45_Ac8 
                 10.0 
                 76.0 
                 8.5 
               
               
                   
                 3.0 
                 69.5 
                 12.0 
               
               
                   
                 1.0 
                 43.5 
                 19.1 
               
               
                 hEx45_Ac9 
                 10.0 
                 73.7 
                 6.0 
               
               
                   
                 3.0 
                 62.5 
                 9.2 
               
               
                   
                 1.0 
                 47.3 
                 8.3 
               
               
                   
                 0.1 
                 20.0 
                   
               
               
                 hEx45_Ac10 
                 10.0 
                 53.0 
                 0.0 
               
               
                   
                 3.0 
                 56.5 
                 10.6 
               
               
                   
                 1.0 
                 35.5 
                 0.7 
               
               
                 hEx45_Ac11 
                 10.0 
                 54.5 
                 2.1 
               
               
                   
                 3.0 
                 53.0 
                 1.4 
               
               
                   
                 1.0 
                 34.0 
                 4.2 
               
               
                 hEx45_Ac12 
                 10.0 
                 52.0 
                 21.2 
               
               
                   
                 3.0 
                 40.0 
                 14.1 
               
               
                   
                 1.0 
                 26.5 
                 10.6 
               
               
                 No PMO 
                 0 
                 10.5 
                 6.4 
               
               
                   
               
            
           
         
       
     
     Table 20B illustrates exon skipping activity of PMOs (30mer) targeting DMD exon 44 in transfected primary human skeletal muscle cells. 
     
       
         
           
               
               
               
            
               
                   
               
               
                   
                 PMO conc 
                 % Skipping (skipped/total) 
               
            
           
           
               
               
               
               
            
               
                   
                 uM 
                 AVG 
                 STDEV 
               
               
                   
               
            
           
           
               
               
               
               
            
               
                 hEx44_Ac0 
                 10 
                 83.8 
                 11.3 
               
               
                   
                 3 
                 79.7 
                 3.5 
               
               
                   
                 1 
                 67.5 
                 7.8 
               
               
                   
                 0.1 
                 31.5 
                 0.7 
               
               
                 hEx44_Ac1 
                 10 
                 77.7 
                 8.3 
               
               
                   
                 3 
                 79.5 
                 0.7 
               
               
                   
                 1 
                 68.3 
                 8.5 
               
               
                   
                 0.1 
                 32.0 
                   
               
               
                 hEx44_Ac2 
                 10 
                 88.7 
                 4.5 
               
               
                   
                 3 
                 96.0 
                 7.1 
               
               
                   
                 1 
                 70.0 
                 13.2 
               
               
                   
                 0.1 
                 31.0 
                   
               
               
                 hEx44_Ac3 
                 10 
                 75.0 
                 14.1 
               
               
                   
                 3 
                 89.0 
                   
               
               
                   
                 1 
                 62.0 
                 8.5 
               
               
                   
                 0.1 
                 26.0 
                   
               
               
                 hEx44_Ac4 
                 10 
                 84.0 
                 17.0 
               
               
                   
                 3 
                 88.0 
                   
               
               
                   
                 1 
                 67.0 
                 15.6 
               
               
                   
                 0.1 
                 23.0 
                   
               
               
                 hEx44_Ac5 
                 10 
                 63.0 
                 0.0 
               
               
                   
                 3 
                 68.0 
                   
               
               
                   
                 1 
                 54.0 
                 8.5 
               
               
                   
                 0.1 
                 18.0 
                   
               
               
                 hEx44_Ac6 
                 10 
                 74.0 
                 12.7 
               
               
                   
                 3 
                 81.0 
                   
               
               
                   
                 1 
                 58.5 
                 17.7 
               
               
                   
                 0.1 
                 20.0 
                   
               
               
                 hEx44_Ac7 
                 10 
                 84.3 
                 19.5 
               
               
                   
                 3 
                 85.0 
                 4.2 
               
               
                   
                 1 
                 59.3 
                 13.0 
               
               
                   
                 0.1 
                 23.0 
                   
               
               
                 hEx44_Ac8 
                 10 
                 76.0 
                 0.0 
               
               
                   
                 3 
                 70.0 
                   
               
               
                   
                 1 
                 53.5 
                 2.1 
               
               
                   
                 0.1 
                 27.0 
                   
               
               
                 hEx44_Ac9 
                 10 
                 76.5 
                 2.1 
               
               
                   
                 3 
                 73.0 
                   
               
               
                   
                 1 
                 59.0 
                 15.6 
               
               
                   
                 0.1 
                 32.0 
                   
               
               
                 hEx44_Ac10 
                 10 
                 85.0 
                 18.4 
               
               
                   
                 3 
                 79.0 
                   
               
               
                   
                 1 
                 45.5 
                 6.4 
               
               
                   
                 0.1 
                 23.0 
                   
               
               
                 hEx44_Ac14 
                 10 
                 86.5 
                 19.1 
               
               
                   
                 3 
                 80.0 
                 11.8 
               
               
                   
                 1 
                 62.0 
                 9.0 
               
               
                   
                 0.1 
                 31.5 
                 0.7 
               
               
                 No PMO 
                   
                 8.3 
                 3.8 
               
               
                   
               
            
           
         
       
     
       FIG. 15  illustrates exon skipping activity of different lengths of hEx45_Ac9 PMOs in transfected primary human skeletal muscle cells. 
     Example 10. Synthesis and Purification of Human TfR1 PMO Conjugates 
     An anti-human transferrin receptor antibody was produced. PMOs (28-mers) were synthesized by GeneTools. Antibody (10 mg/ml) in borate buffer (25 mM sodium tetraborate, 25 mM NaCl, 1 mM Diethylene triamine pentaacetic acid, pH 8.0) was reduced by adding 4 equivalents of tris(2-carboxyethyl)phosphine (TCEP) in water and incubating at 37° C. for 4 hours. 4(N-Maleimidomethyl)cyclohexanecarboxylic acid N-hydroxysuccinimide ester (SMCC) was coupled to the primary amine on the 3′ end of the PMO by incubating the PMO (50 mg/ml) in DMSO with 10 equivalents of SMCC (10 mg/ml) in DMSO for one hour. Unconjugated SMCC was removed by ultrafiltration using Amicon Ultra-15 centrifugal filter units with a MWCO of 3 kDa. The PMO-SMCC was washed three times with acetate buffer (10 mM sodium acetate, pH 6.0) and used immediately. The reduced antibody was mixed with 2.25 equivalents of PMO-SMCC and incubated overnight at 4° C. The pH of the reaction mixture was then reduced to 7.5 and 8 equivalents of N-Ethylmaleimide was added to the mixture at room temperature for 30 minutes to quench unreacted cysteines. Analysis of the reaction mixture by hydrophobic interaction chromatography (HIC) method-2 showed antibody-PMO conjugates along with unreacted antibody and PMO. 
     The reaction mixture was purified with an AKTA Explorer FPLC using HIC method-1. Dependent on the conjugate, fractions containing either conjugates with a drug to antibody ratio of one (DAR 1), two (DAR 2), and three (DAR 3), or fractions containing conjugates with a drug to antibody ratio of 3+(DAR 3+), or 4+(DAR 4+) were combined and concentrated with Amicon Ultra-15 centrifugal filter units with a MWCO of 50 kDa. Concentrated conjugates were buffer exchanged with PBS (pH 7.4) using Amicon Ultra-15 centrifugal filter units prior to analysis. 
     Hydrophobic Interaction Chromatography (HIC) Method-1.
     1. Column: GE, HiScreen Butyl HP, 4.7 ml   2. Solvent A: 50 mM phosphate buffer, 0.7M Ammonium Sulfate, pH 7.0; Solvent B: 80% 50 mM phosphate buffer, 20% IPA, pH 7.0; Flow Rate: 1.0 ml/min   3. Gradient:   

     
       
         
           
               
               
               
               
               
             
               
                   
               
             
            
               
                   
                 a. 
                 % A 
                 % B 
                 Column Volume  
               
               
                   
                 b. 
                 100 
                  0 
                  1 
               
               
                   
                 c. 
                  70 
                  30 
                 25 
               
               
                   
                 d. 
                  0 
                 100 
                  1 
               
               
                   
                 e. 
                  0 
                 100 
                  2 
               
               
                   
               
            
           
         
       
     
     Binding of hTfR1.mAb-PMO Conjugates to Human Transferrin Receptor 
     Antibody conjugate (AOC) binding was measured by ELISA. Recombinant human Transferrin Receptor (Sino Biological 11020-HO7H) was coated onto high bind plates (Costar 3690) at 1 ng/uL in PBS overnight. Plates were washed and AOC or mAb samples were added at concentrations up to 10 nM. Color was developed through HRP conjugated secondary antibody (Jackson Immunoresearch 109-035-006) and TMB substrate (ThermoFisher 34028) stopped with 2N sulfuric acid. Kd was determined using GraphPad Prism. 
       FIG. 16  illustrates binding of hTfR1.mAb-PMO conjugates to human Transferrin Receptor in vitro. 
     Activity of TfR1 mAb-PMO Conjugates in Primary Human Skeletal Muscle Cells 
     Primary, pre-differentiated human skeletal muscle cells (Gibco, # A11440) were plated on collagen Type 1 coated 24-well plates (Gibco, #1970788) in DMEM supplemented with 2% horse serum and 1×ITS (Gibco, #1933286) according to the manufacturer&#39;s instructions. Cells were grown in 37° C.+5% CO 2  for 2 days to establish myotubes. Immortalized human skeletal muscle cells from healthy donors (Myology Institute Paris) were plated on collagen Type 1 coated 24-well plates (Gibco, #1970788) in Skeletal Muscle Cell Growth medium (Promocell, C-23160) supplemented with 5% FBS. After myoblasts reached confluency, myotube formation was induced in differentiation medium containing DMEM supplemented with gentamycin (50 ug/ml) (Invitrogen, 15750-045) and insulin (10 ug/ml) (sigma, 91077). Myotubes were then treated with defined concentrations of AOCs in the respective medium. Cell were harvested 72 hours after treatment by aspirating the culture medium, followed by addition of 300 ul TRIZOL per well. RNA isolation and quantification of DMD exon skipping was performed as detailed in example 9. 
       FIG. 17  illustrates exon skipping activity of hTfR1.mAb-PMO (28-mer) conjugates in primary human skeletal muscle cells. 
       FIG. 18  illustrates exon skipping activity of hTfR1.mAb-PMO conjugates in myotubes of primary and immortalized human skeletal muscle cells. 
     While preferred embodiments of the present disclosure have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the disclosure. It should be understood that various alternatives to the embodiments of the disclosure described herein may be employed in practicing the disclosure. It is intended that the following claims define the scope of the disclosure and that methods and structures within the scope of these claims and their equivalents be covered thereby.