Patent Publication Number: US-2023159927-A1

Title: Chromatin remodelers to enhance targeted gene activation

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims priority to U.S. Provisional Patent Application No. 63/022,174, filed May 8, 2020, and U.S. Provisional Patent Application No. 63/094,158 d Oct. 20, 2020, each of which is incorporated herein by reference in its entirety. 
    
    
     STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH 
     This invention was made with government support under grant EFMA-1830957 awarded by the National Science Foundation, grant SP5243390 awarded by the Defense Advanced Research Projects Agency, and grant U01Al146356 awarded by the National Institutes of Health. The government has certain rights in the invention. 
    
    
     FIELD 
     This disclosure relates to compositions and methods for the targeted activation of genes. 
     INTRODUCTION 
     Targeted activation of endogenous genes with synthetic transcription factors or epigenome editors made from DNA-targeting systems such as zinc finger proteins, TALEs, and CRISPR-Cas systems, are broadly useful for gene therapy, regenerative medicine, and programming stem cell differentiation. However, in some cases the potency or specificity of gene activation is insufficient to generate the desired phenotype or biological effect. There is a need for improved systems for activating expression of a specific gene. 
     SUMMARY 
     In an aspect, the disclosure relates to a fusion protein comprising at least two heterologous polypeptide domains, wherein the first polypeptide domain comprises a DNA binding protein and the second polypeptide domain comprises a modulator of chromatin structure. In some embodiments, the fusion protein further comprises a third polypeptide domain. In some embodiments, the first polypeptide domain comprises a CRISPR-associated (Cas) protein, a TALE, or a zinc finger protein. In some embodiments, the Cas protein comprises at least one amino acid mutation that eliminates nuclease activity of the Cas protein. In some embodiments, the Cas protein comprises a Cas9 protein. In some embodiments, the Cas9 protein is nuclease-deficient dCas9 and comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 20 or 21 or is encoded by a polynucleotide comprising a sequence having at least 75% identity to SEQ ID NO: 22 or 23. In some embodiments, the modulator of chromatin structure comprises a nucleosome rearranging protein. In some embodiments, the modulator of chromatin structure comprises the SS18 subunit of the BAF chromatin remodeling complex or a fragment thereof or a variant thereof. In some embodiments, the SS18 subunit comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 37. In some embodiments, the third polypeptide domain comprises a transcriptional activator domain. In some embodiments, the transcriptional activator domain comprises VP64, VPH, VPR, p65, TET1, or p300, or a combination thereof or a fragment thereof or a variant thereof. In some embodiments, the VP64 comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 91. In some embodiments, the TET1 comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 93. In some embodiments, the VPH comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 39, In some embodiments, the VPR comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 41. In some embodiments, the p300 comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 33 or 34. In some embodiments, the fusion protein comprises one or more second polypeptide domain(s). In some embodiments, the one or more second polypeptide domain(s) is fused to the C-terminus or the N-terminus of the first polypeptide domain, or a combination thereof. In some embodiments, the N-terminus of the second polypeptide is operably linked to the C-terminus of the first polypeptide domain, or the C-terminus of the second polypeptide is operably linked to the N-terminus of the first polypeptide domain. In some embodiments, the fusion protein comprises one or more third polypeptide domain(s). In some embodiments, the one or more third polypeptide domain is fused to the C-terminus or the N-terminus of the first polypeptide domain, or a combination thereof. In some embodiments, the N-terminus of the third polypeptide is operably linked to the C-terminus of the first polypeptide domain, or the C-terminus of the third polypeptide is operably linked to the N-terminus of the first polypeptide domain. In some embodiments, the first polypeptide domain comprises dCas9, the second polypeptide domain comprises SS18, and the third polypeptide domain comprises VPH. In some embodiments, the fusion protein comprises VPH-dCas9-SS18 or SS18-dCas9-VPH or variants thereof. In some embodiments, the fusion protein comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 64 or 66. In some embodiments, the first polypeptide domain comprises dCas9, the second polypeptide domain comprises SS18, and the third polypeptide domain comprises VPR. In some embodiments, the fusion protein comprises VPR-dCas9-SS18 or SS18-dCas9-VPR or variants thereof. In some embodiments, the first polypeptide domain comprises dCas9, the second polypeptide domain comprises SS18, and the third polypeptide domain comprises p300, In some embodiments, the fusion protein comprises p300-dCas9-SS18 or SS18-dCas9-p300 or variants thereof. In some embodiments, the first polypeptide domain comprises dCas9, the second polypeptide domain comprises SS18, and the third polypeptide domain comprises VP64. In some embodiments, the fusion protein comprises VP64-dCas9-SS18 or SS18-dCas9-VP64 or variants thereof. In some embodiments, the fusion protein activates transcription of a target gene. In some embodiments, the fusion protein increases the level of mRNA expression of a target gene in a cell containing the fusion protein relative to a control. In some embodiments, the level of mRNA expression of the target gene is increased at least 5-fold, at least 50-fold, at least 100-fold, at least 1,000-fold, at least 10,000-fold, or at least 20,000-fold relative to a control. In some embodiments, the level of mRNA expression of the target gene is increased by 5-fold to 10,000-fold, 5-fold to 30,000-fold, 5-fold to 50,000-fold, 5-fold to 100,000-fold, 10,000-fold to 30,000-fold, 20,000-fold to 30,000-fold, 15,000-fold to 25,000-fold, 1,000-fold to 50,000-fold, or 1.000-fold to 100,000-fold relative to a control. In some embodiments, the control is the level of mRNA expression of the target gene in a cell not containing the fusion protein. In some embodiments, the target gene is gamma globin genes 1 and 2 (HBG1/2). 
     In a further aspect, the disclosure relates to DNA Targeting System. The DNA Targeting System may include (a) a fusion protein as detailed herein, wherein the first polypeptide domain comprises a zinc finger protein or a TALE; or (b) a gRNA and a fusion protein as detailed herein, wherein the first polypeptide domain comprises a Cas protein, and wherein the gRNA targets a target gene. In some embodiments, gRNA targets a regulatory region of the target gene. In some embodiments, the regulatory region is a promoter sequence of the target gene. Another aspect of the disclosure provides DNA Targeting System comprising a gRNA that recruits a modulator of chromatin structure to a target sequence. In some embodiments, the modulator of chromatin structure comprises the SS18 subunit of the BAF chromatin remodeling complex. In some embodiments, the gRNA is encoded by or binds to a target sequence selected from SEQ ID NOs: 43-48, a complement thereof, a truncation thereof, or a variant thereof, or the gRNA is encoded by or binds to a target sequence having at least 70% sequence identity to a sequence selected from SEQ ID NOs: 43-48, a complement thereof, a truncation thereof, or a variant thereof. In some embodiments, the gRNA comprises a polynucleotide sequence selected from SEQ ID NOs: 49-54, a complement thereof, a truncation thereof, or a variant thereof, or the gRNA comprises a polynucleotide having at least 70% sequence identity to a sequence selected from SEQ ID NOs: 49-54, a complement thereof, a truncation thereof, or a variant thereof. 
     Another aspect of the disclosure provides a method of increasing expression of a target gene in a cell. The method may include contacting the cell with a fusion protein as detailed herein or a DNA Targeting system as detailed herein. In some embodiments, the target gene is gamma globin genes 1 and 2 (HBG1/2). 
     Another aspect of the disclosure provides a gRNA encoded by or binding to a target sequence selected from SEQ ID NOs: 43-48, a complement thereof, a truncation thereof, or a variant thereof, or comprising a polynucleotide sequence selected from SEQ ID NOs: 49-54, a complement thereof, a truncation thereof, or a variant thereof. 
     The disclosure provides for other aspects and embodiments that will be apparent in light of the following detailed description and accompanying figures. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG.  1 A  shows the effect of indicated dCas9 activator fusions on HBG112 expression when targeted to the HBG1/2 promoter (with 2 gRNAs) or to the distal HS2 enhancer of the globin locus (with 4 gRNAs) in HEK293T cells, evaluated by RT-qPCR (n=2).  FIG.  1 B  shows a schematic of the indicated dCas9 activator fusions (NLS, nuclear localization signal).  FIG.  1 C  is a Western blot showing that the expression of the indicated dCas9 fusions is similar. Asterisk denotes degradation products. 
         FIG.  2    shows the effect of indicated dCas9 activator fusions on HBG1/2 expression when targeted to the HBG112 promoter (with 2 gRNAs) in HEK293T cells, evaluated by RT-qPCR (n=2). 
         FIG.  3    shows the effect of indicated dCas9 activator fusions on HBG1/2 expression when targeted to the HBG1/2 promoter (with 2 gRNAs) (left) and the expression level of the dCas9 activator fusions (right) in HEK293T cells, evaluated by RT-qPCR (n=2). 
         FIG.  4    shows the effect of indicated dCas9 activator fusions on HBG1/2 expression when targeted to the HBG1/2 promoter (with 2 gRNAs) (left) and the expression level of the dCas9 activator fusions (right) in HEK293T cells, evaluated by RT-qPCR (n=2). 
     
    
    
     DETAILED DESCRIPTION 
     Described herein are fusion proteins for the targeted activation of genes as well as compositions and methods comprising the same. As detailed herein, it was demonstrated that combining modulators of chromatin structure, such as proteins that rearrange nucleosomes and/or cause movement of DNA in relation to the nucleosomes, with activator domains can lead to more potent gene activation in human cells. Remodelers of chromatin structure can cooperate with co-recruited transcriptional activation domains to more robustly activate target gene expression. The fusion protein may include at least two heterologous polypeptide domains. The first polypeptide domain includes a DNA binding protein, such as a zinc finger protein, a TALE, or a Cas9 protein, that targets the fusion protein for binding to a specific DNA sequence. The second polypeptide domain includes a modulator of chromatin structure, such as a nucleosome rearranging domain. The fusion protein may further include a third polypeptide domain, the third polypeptide domain including a transcriptional activator domain. The fusion protein may be incorporated into a DNA Targeting System and may be used to activate expression of a target gene in a cell. 
     1. Definitions 
     Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting. 
     The terms “comprise(s),” “include(s),” “having,” “has,” “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms “a,” “and,” and “the” include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments “comprising,” “consisting of,” and “consisting essentially of,” the embodiments or elements presented herein, whether explicitly set forth or not. 
     For the recitation of numeric ranges herein, each intervening number there between with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated. 
     The term “about” or “approximately” as used herein as applied to one or more values of interest, refers to a value that is similar to a stated reference value, or within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, such as the limitations of the measurement system. In certain aspects, the term “about” refers to a range of values that fall within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value). Alternatively, “about” can mean within 3 or more than 3 standard deviations, per the practice in the art. Alternatively, such as with respect to biological systems or processes, the term “about” can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value. 
     “Adeno-associated virus” or “AAV” as used interchangeably herein refers to a small virus belonging to the genus Dependovirus of the Parvoviridae family that infects humans and some other primate species. AAV is not currently known to cause disease and consequently the virus causes a very mild immune response. 
     “Amino acid” as used herein refers to naturally occurring and non-natural synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code. Amino acids can be referred to herein by either their commonly known three-letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Amino acids include the side chain and polypeptide backbone portions. 
     “Binding region” as used herein refers to the region within a target region that is recognized and bound by the CRISPR/Cas-based gene editing system. 
     “Clustered Regularly Interspaced Short Palindromic Repeats” and “CRISPRs”, as used interchangeably herein, refers to loci containing multiple short direct repeats that are found in the genomes of approximately 40% of sequenced bacteria and 90% of sequenced archaea. 
     “Coding sequence” or “encoding nucleic acid” as used herein means the nucleic acids (RNA or DNA molecule) that comprise a nucleotide sequence which encodes a protein. The coding sequence can further include initiation and termination signals operably linked to regulatory elements including a promoter and polyadenylation signal capable of directing expression in the cells of an individual or mammal to which the nucleic acid is administered. The coding sequence may be codon optimized. 
     “Complement” or “complementary” as used herein means a nucleic acid can mean Watson-Crick (e.g., A-T/U and C-G) or Hoogsteen base pairing between nucleotides or nucleotide analogs of nucleic acid molecules. “Complementarity” refers to a property shared between two nucleic acid sequences, such that when they are aligned antiparallel to each other, the nucleotide bases at each position will be complementary. 
     The terms “control,” “reference level,” and “reference” are used herein interchangeably. The reference level may be a predetermined value or range, which is employed as a benchmark against which to assess the measured result. “Control group” as used herein refers to a group of control subjects. The predetermined level may be a cutoff value from a control group. The predetermined level may be an average from a control group. Cutoff values (or predetermined cutoff values) may be determined by Adaptive Index Model (AIM) methodology. Cutoff values (or predetermined cutoff values) may be determined by a receiver operating curve (ROC) analysis from biological samples of the patient group, ROC analysis, as generally known in the biological arts, is a determination of the ability of a test to discriminate one condition from another, e.g., to determine the performance of each marker in identifying a patient having CRC. A description of ROC analysis is provided in P. J. Heagerty et al. (Biometrics 2000, 56, 337-44), the disclosure of which is hereby incorporated by reference in its entirety. Alternatively, cutoff values may be determined by a quartile analysis of biological samples of a patient group. For example, a cutoff value may be determined by selecting a value that corresponds to any value in the 25th-75th percentile range, preferably a value that corresponds to the 25th percentile, the 50th percentile or the 75th percentile, and more preferably the 75th percentile. Such statistical analyses may be performed using any method known in the art and can be implemented through any number of commercially available software packages (e.g., from Analyse-it Software Ltd., Leeds, UK; StataCorp LP, College Station, Tex.; SAS Institute Inc., Cary, N.C.). The healthy or normal levels or ranges for a target or for a protein activity may be defined in accordance with standard practice. A control may be a subject or cell without a fusion protein as detailed herein, A control may be a subject, or a sample therefrom, whose disease state is known. The subject, or sample therefrom, may be healthy, diseased, diseased prior to treatment, diseased during treatment, or diseased after treatment, or a combination thereof. 
     “Correcting”, “gene editing,” and “restoring” as used herein refers to changing a mutant gene that encodes a dysfunctional protein or truncated protein or no protein at all, such that a full-length functional or partially full-length functional protein expression is obtained. Correcting or restoring a mutant gene may include replacing the region of the gene that has the mutation or replacing the entire mutant gene with a copy of the gene that does not have the mutation with a repair mechanism such as homology-directed repair (HDR). Correcting or restoring a mutant gene may also include repairing a frameshift mutation that causes a premature stop codon, an aberrant splice acceptor site or an aberrant splice donor site, by generating a double stranded break in the gene that is then repaired using non-homologous end joining (NHEJ). NHEJ may add or delete at least one base pair during repair which may restore the proper reading frame and eliminate the premature stop codon. Correcting or restoring a mutant gene may also include disrupting an aberrant splice acceptor site or splice donor sequence. Correcting or restoring a mutant gene may also include deleting a non-essential gene segment by the simultaneous action of two nucleases on the same DNA strand in order to restore the proper reading frame by removing the DNA between the two nuclease target sites and repairing the DNA break by NHEJ. 
     “Donor DNA”, “donor template,” and “repair template” as used interchangeably herein refers to a double-stranded DNA fragment or molecule that includes at least a portion of the gene of interest. The donor DNA may encode a full-functional protein or a partially functional protein. 
     “Enhancer” as used herein refers to non-coding DNA sequences containing multiple activator and repressor binding sites. Enhancers range from 200 bp to 1 kb in length and may be either proximal, 5′ upstream to the promoter or within the first intron of the regulated gene, or distal, in introns of neighboring genes or intergenic regions far away from the locus. Through DNA looping, active enhancers contact the promoter dependently of the core DNA binding motif promoter specificity. 4 to 5 enhancers may interact with a promoter. Similarly, enhancers may regulate more than one gene without linkage restriction and may “skip” neighboring genes to regulate more distant ones. Transcriptional regulation may involve elements located in a chromosome different to one where the promoter resides. Proximal enhancers or promoters of neighboring genes may serve as platforms to recruit more distal elements. 
     “Frameshift” or “frameshift mutation” as used interchangeably herein refers to a type of gene mutation wherein the addition or deletion of one or more nucleotides causes a shift in the reading frame of the codons in the mRNA. The shift in reading frame may lead to the alteration in the amino acid sequence at protein translation, such as a missense mutation or a premature stop codon. 
     “Functional” and “full-functional” as used herein describes protein that has biological activity. A “functional gene” refers to a gene transcribed to mRNA, which is translated to a functional protein. 
     “Fusion protein” as used herein refers to a chimeric protein created through the joining of two or more genes that originally coded for separate proteins. The translation of the fusion gene results in a single polypeptide with functional properties derived from each of the original proteins. 
     “Genetic construe” as used herein refers to the DNA or RNA molecules that comprise a polynucleotide that encodes a protein. The coding sequence includes initiation and termination signals operably linked to regulatory elements including a promoter and polyadenylation signal capable of directing expression in the cells of the individual to whom the nucleic acid molecule is administered. As used herein, the term “expressible form” refers to gene constructs that contain the necessary regulatory elements operable linked to a coding sequence that encodes a protein such that when present in the cell of the individual, the coding sequence will be expressed. 
     “Genome editing” or “gene editing” as used herein refers to changing a gene. Genome editing may include correcting or restoring a mutant gene or adding additional mutations. Genome editing may include knocking out a gene, such as a mutant gene or a normal gene. Genome editing may be used to treat disease or, for example, enhance muscle repair, by changing the gene of interest. In some embodiments, the compositions and methods detailed herein are for use in somatic cells and not germ line cells. 
     The term “heterologous” as used herein refers to a nucleic acid or protein comprising two or more subsequences that are not found in the same relationship to each other in nature. For instance, a nucleic acid that is recombinantly produced typically has two or more sequences from unrelated genes synthetically arranged to make a new functional nucleic acid, for example, a promoter from one source and a coding region from another source. The two nucleic acids are thus heterologous to each other in this context. When added to a cell, the recombinant nucleic acids would also be heterologous to the endogenous genes of the cell. Thus, in a chromosome, a heterologous nucleic acid would include a non-native (non-naturally occurring) nucleic acid that has integrated into the chromosome, or a non-native (non-naturally occurring) extrachromosomal nucleic acid. Similarly, a heterologous protein indicates that the protein comprises two or more subsequences that are not found in the same relationship to each other in nature (for example, a “fusion protein,” where the two subsequences are encoded by a single nucleic acid sequence). 
     “Homology-directed repair” or “HDR” as used interchangeably herein refers to a mechanism in cells to repair double strand DNA lesions when a homologous piece of DNA is present in the nucleus, mostly in G2 and S phase of the cell cycle. HDR uses a donor DNA template to guide repair and may be used to create specific sequence changes to the genome, including the targeted addition of whole genes. If a donor template is provided along with the CRISPR/Cas9-based gene editing system, then the cellular machinery will repair the break by homologous recombination, which is enhanced several orders of magnitude in the presence of DNA cleavage. When the homologous DNA piece is absent, non-homologous end joining may take place instead. 
     “Identical” or “identity” as used herein in the context of two or more polynucleotide or polypeptide sequences means that the sequences have a specified percentage of residues that are the same over a specified region. The percentage may be calculated by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of different lengths or the alignment produces one or more staggered ends and the specified region of comparison includes only a single sequence, the residues of single sequence are included in the denominator but not the numerator of the calculation. When comparing DNA and RNA, thymine (T) and uracil (U) may be considered equivalent. Identity may be performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0. 
     “Mutant gene” or “mutated gene” as used interchangeably herein refers to a gene that has undergone a detectable mutation. A mutant gene has undergone a change, such as the loss, gain, or exchange of genetic material, which affects the normal transmission and expression of the gene. A “disrupted gene” as used herein refers to a mutant gene that has a mutation that causes a premature stop codon. The disrupted gene product is truncated relative to a full-length undisrupted gene product. 
     “Non-homologous end joining (NHEJ) pathway” as used herein refers to a pathway that repairs double-strand breaks in DNA by directly ligating the break ends without the need for a homologous template. The template-independent re-ligation of DNA ends by NHEJ is a stochastic, error-prone repair process that introduces random micro-insertions and micro-deletions (indels) at the DNA breakpoint. This method may be used to intentionally disrupt, delete, or alter the reading frame of targeted gene sequences. NHEJ typically uses short homologous DNA sequences called microhomologies to guide repair. These microhomologies are often present in single-stranded overhangs on the end of double-strand breaks. When the overhangs are perfectly compatible, NHEJ usually repairs the break accurately, yet imprecise repair leading to loss of nucleotides may also occur, but is much more common when the overhangs are not compatible. “Nuclease mediated NHEJ” as used herein refers to NHEJ that is initiated after a nuclease cuts double stranded DNA. 
     “Normal gene” as used herein refers to a gene that has not undergone a change, such as a loss, gain, or exchange of genetic material. The normal gene undergoes normal gene transmission and gene expression. For example, a normal gene may be a wild-type gene. 
     “Nucleic acid” or “oligonucleotide” or “polynucleotide” as used herein means at least two nucleotides covalently linked together. The depiction of a single strand also defines the sequence of the complementary strand, Thus, a polynucleotide also encompasses the complementary strand of a depicted single strand. Many variants of a polynucleotide may be used for the same purpose as a given polynucleotide. Thus, a polynucleotide also encompasses substantially identical polynucleotides and complements thereof. A single strand provides a probe that may hybridize to a target sequence under stringent hybridization conditions. Thus, a polynucleotide also encompasses a probe that hybridizes under stringent hybridization conditions. Polynucleotides may be single stranded or double stranded or may contain portions of both double stranded and single stranded sequence. The polynucleotide can be nucleic acid, natural or synthetic, DNA, genomic DNA, cDNA, RNA, or a hybrid, where the polynucleotide can contain combinations of deoxyribo- and ribs-nucleotides, and combinations of bases including, for example, uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine, and isoguanine. Polynucleotides can be obtained by chemical synthesis methods or by recombinant methods. 
     “Open reading frame” refers to a stretch of codons that begins with a start codon and ends at a stop codon. In eukaryotic genes with multiple exons, introns are removed, and exons are then joined together after transcription to yield the final mRNA for protein translation. An open reading frame may be a continuous stretch of codons. In some embodiments, the open reading frame only applies to spliced mRNAs, not genomic DNA, for expression of a protein. 
     “Operably linked” as used herein means that expression of a gene is under the control of a promoter with which it is spatially connected. A promoter may be positioned 5′ (upstream) or 3′ (downstream) of a gene under its control. The distance between the promoter and a gene may be approximately the same as the distance between that promoter and the gene it controls in the gene from which the promoter is derived. As is known in the art, variation in this distance may be accommodated without loss of promoter function. Nucleic acid or amino acid sequences are “operably linked” (or “operatively linked”) when placed into a functional relationship with one another. For instance, a promoter or enhancer is operably linked to a coding sequence if it regulates, or contributes to the modulation of, the transcription of the coding sequence. Operably linked DNA sequences are typically contiguous, and operably linked amino acid sequences are typically contiguous and in the same reading frame. However, since enhancers generally function when separated from the promoter by up to several kilobases or more and intronic sequences may be of variable lengths, some polynucleotide elements may be operably linked but not contiguous. Similarly, certain amino acid sequences that are non-contiguous in a primary polypeptide sequence may nonetheless be operably linked due to, for example folding of a polypeptide chain. With respect to fusion polypeptides, the terms “operatively linked” and “operably linked” can refer to the fact that each of the components performs the same function in linkage to the other component as it would if it were not so linked. 
     “Partially-functional” as used herein describes a protein that is encoded by a mutant gene and has less biological activity than a functional protein but more than a non-functional protein. 
     A “peptide” or “polypeptide” is a linked sequence of two or more amino acids linked by peptide bonds. The polypeptide can be natural, synthetic, or a modification or combination of natural and synthetic. Peptides and polypeptides include proteins such as binding proteins, receptors, and antibodies. The terms “polypeptide”, “protein,” and “peptide” are used interchangeably herein. “Primary structure” refers to the amino acid sequence of a particular peptide. “Secondary structure” refers to locally ordered, three dimensional structures within a polypeptide. These structures are commonly known as domains, for example, enzymatic domains, extracellular domains, transmembrane domains, pore domains, and cytoplasmic tail domains. “Domains” are portions of a polypeptide that form a compact unit of the polypeptide and are typically 15 to 350 amino acids long. Exemplary domains include domains with enzymatic activity or ligand binding activity. Typical domains are made up of sections of lesser organization such as stretches of beta-sheet and alpha-helices. “Tertiary structure” refers to the complete three-dimensional structure of a polypeptide monomer. “Quaternary structure” refers to the three-dimensional structure formed by the noncovalent association of independent tertiary units. A “motif” is a portion of a polypeptide sequence and includes at least two amino acids. A motif may be 2 to 20, 2 to 15, or 2 to 10 amino acids in length. In some embodiments, a motif includes 3, 4, 5, 6, or 7 sequential amino acids. A domain may be comprised of a series of the same type of motif. 
     “Premature stop codon” or “out-of-frame stop codon” as used interchangeably herein refers to nonsense mutation in a sequence of DNA, which results in a stop codon at location not normally found in the wild-type gene. A premature stop codon may cause a protein to be truncated or shorter compared to the full-length version of the protein. 
     “Promoter” as used herein means a synthetic or naturally derived molecule which is capable of conferring, activating or enhancing expression of a nucleic acid in a cell. A promoter may comprise one or more specific transcriptional regulatory sequences to further enhance expression and/or to alter the spatial expression and/or temporal expression of same. A promoter may also comprise distal enhancer or repressor elements, which may be located as much as several thousand base pairs from the start site of transcription. A promoter may be derived from sources including viral, bacterial, fungal, plants, insects, and animals. A promoter may regulate the expression of a gene component constitutively, or differentially with respect to cell, the tissue or organ in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, pathogens, metal ions, or inducing agents. Representative examples of promoters include the bacteriophage T7 promoter, bacteriophage T3 promoter, SP6 promoter, lac operator-promoter, tac promoter, SV40 late promoter, SV40 early promoter, RSV-LTR promoter, CMV IE promoter, SV40 early promoter or SV40 late promoter, human U6 (hU6) promoter, and CMV IE promoter. Promoters that target muscle-specific stem cells may include the CK8 promoter, the Spc5-12 promoter, and the MHCK7 promoter. 
     The term “recombinant” when used with reference to, for example, a cell, nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein, or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found within the native (naturally occurring) form of the cell or express a second copy of a native gene that is otherwise normally or abnormally expressed, under expressed, or not expressed at all. 
     “Sample” or “test sample” as used herein can mean any sample in which the presence and/or level of a target is to be detected or determined or any sample comprising a DNA targeting or gene editing system or component thereof as detailed herein. Samples may include liquids, solutions, emulsions, or suspensions. Samples may include a medical sample. Samples may include any biological fluid or tissue, such as blood, whole blood, fractions of blood such as plasma and serum, muscle, interstitial fluid, sweat, saliva, urine, tears, synovial fluid, bone marrow, cerebrospinal fluid, nasal secretions, sputum, amniotic fluid, bronchoalveolar lavage fluid, gastric lavage, emesis, fecal matter, lung tissue, peripheral blood mononuclear cells, total white blood cells, lymph node cells, spleen cells, tonsil cells, cancer cells, tumor cells, bile, digestive fluid, skin, or combinations thereof. In some embodiments, the sample comprises an aliquot. In other embodiments, the sample comprises a biological fluid. Samples can be obtained by any means known in the art. The sample can be used directly as obtained from a patient or can be pre-treated, such as by filtration, distillation, extraction, concentration, centrifugation, inactivation of interfering components, addition of reagents, and the like, to modify the character of the sample in some manner as discussed herein or otherwise as is known in the art. 
     “Subject” and “patient” as used herein interchangeably refers to any vertebrate, including, but not limited to, a mammal that wants or is in need of the herein described compositions or methods. The subject may be a human or a non-human. The subject may be a vertebrate. The subject may be a mammal. The mammal may be a primate or a non-primate. The mammal can be a non-primate such as, for example, cow, pig, camel, llama, hedgehog, anteater, platypus, elephant, alpaca, horse, goat, rabbit, sheep, hamsters, guinea pig, cat, dog, rat, and mouse. The mammal can be a primate such as a human. The mammal can be a non-human primate such as, for example, monkey, cynomolgous monkey, rhesus monkey, chimpanzee, gorilla, orangutan, and gibbon. The subject may be of any age or stage of development, such as, for example, an adult, an adolescent, or an infant. The subject may be male. The subject may be female. In some embodiments, the subject has a specific genetic marker. The subject may be undergoing other forms of treatment. 
     “Substantially identical” can mean that a first and second amino acid or polynucleotide sequence have at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity over a region of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100 amino acids or nucleotides, respectively. 
     “Target gene” as used herein refers to any nucleotide sequence encoding a known or putative gene product. The target gene may be a mutated gene involved in a genetic disease. The target gene may encode a known or putative gene product that is intended to be corrected or for which its expression is intended to be modulated. In certain embodiments, the target gene is a gamma globin gene. 
     “Target region” as used herein refers to the region of the target gene to which the CRISPR/Cas9-based gene editing or targeting system is designed to bind. 
     “Transgene” as used herein refers to a gene or genetic material containing a gene sequence that has been isolated from one organism and is introduced into a different organism. This non-native segment of DNA may retain the ability to produce RNA or protein in the transgenic organism, or it may alter the normal function of the transgenic organism&#39;s genetic code. The introduction of a transgene has the potential to change the phenotype of an organism. 
     “Transcriptional regulatory elements” or “regulatory elements” refers to a genetic element which can control the expression of nucleic acid sequences, such as activate, enhance, or decrease expression, or alter the spatial and/or temporal expression of a nucleic acid sequence. Examples of regulatory elements include, for example, promoters, enhancers, splicing signals, polyadenylation signals, and termination signals. A regulatory element can be “endogenous,” “exogenous,” or “heterologous” with respect to the gene to which it is operably linked. An “endogenous” regulatory element is one which is naturally linked with a given gene in the genome. An “exogenous” or “heterologous” regulatory element is one which is not normally linked with a given gene but is placed in operable linkage with a gene by genetic manipulation. 
     “Treatment” or “treating” or “treatment” when referring to protection of a subject from a disease, means suppressing, repressing, reversing, alleviating, ameliorating, or inhibiting the progress of disease, or completely eliminating a disease. A treatment may be either performed in an acute or chronic way. The term also refers to reducing the severity of a disease or symptoms associated with such disease prior to affliction with the disease. Preventing the disease involves administering a composition of the present invention to a subject prior to onset of the disease. Suppressing the disease involves administering a composition of the present invention to a subject after induction of the disease but before its clinical appearance. Repressing or ameliorating the disease involves administering a composition of the present invention to a subject after clinical appearance of the disease. 
     As used herein, the term “gene therapy” refers to a method of treating a patient wherein polypeptides or nucleic acid sequences are transferred into cells of a patient such that activity and/or the expression of a particular gene is modulated. In certain embodiments, the expression of the gene is suppressed. In certain embodiments, the expression of the gene is enhanced. In certain embodiments, the temporal or spatial pattern of the expression of the gene is modulated. 
     “Variant” used herein with respect to a polynucleotide means (i) a portion or fragment of a referenced nucleotide sequence; (ii) the complement of a referenced nucleotide sequence or portion thereof; (iii) a nucleic acid that is substantially identical to a referenced nucleic acid or the complement thereof; or (iv) a nucleic acid that hybridizes under stringent conditions to the referenced nucleic acid, complement thereof, or a sequence substantially identical thereto. 
     “Variant” with respect to a peptide or polypeptide refers to a peptide or polypeptide that differs in amino acid sequence by the insertion, deletion, conservative substitution, or non-conservative substitution of amino acids, but retains at least one biological activity. Variant may also mean a protein with an amino acid sequence that is substantially identical to a referenced protein with an amino acid sequence that retains at least one biological activity. Representative examples of “biological activity” include the ability to be bound by a specific antibody or polypeptide, to promote an immune response, to activate or increase transcription, to bind or target a polynucleotide or a polypeptide, rearrange or remodel chromatin, or to catalyze a reaction such as demethylation or acetylation. Variant can mean a functional fragment or truncation thereof. Variant can also mean multiple copies of a polypeptide. The multiple copies can be in tandem or separated by a linker. A conservative substitution of an amino acid, for example, replacing an amino acid with a different amino acid of similar properties (for example, hydrophilicity, degree and distribution of charged regions) is recognized in the art as typically involving a minor change. These minor changes may be identified, in part, by considering the hydropathic index of amino acids, as understood in the art (Kyte et al., J. Mol. Biol. 1982, 157, 105-132). The hydropathic index of an amino acid is based on a consideration of its hydrophobicity and charge. It is known in the art that amino acids of similar hydropathic indexes may be substituted and still retain protein function. In one aspect, amino acids having hydropathic indexes of ±2 are substituted. The hydrophilicity of amino acids may also be used to reveal substitutions that would result in proteins retaining biological function. A consideration of the hydrophilicity of amino acids in the context of a peptide permits calculation of the greatest local average hydrophilicity of that peptide. Substitutions may be performed with amino acids having hydrophilicity values within ±2 of each other. Both the hydrophobicity index and the hydrophilicity value of amino acids are influenced by the particular side chain of that amino acid. Consistent with that observation, amino acid substitutions that are compatible with biological function are understood to depend on the relative similarity of the amino acids, and particularly the side chains of those amino acids, as revealed by the hydrophobicity, hydrophilicity, charge, size, and other properties. 
     “Vector” as used herein means a nucleic acid sequence containing an origin of replication. A vector may be capable of directing the delivery or transfer of a polynucleotide sequence to target cells, where it can be replicated or expressed. A vector may contain an origin of replication, one or more regulatory elements, and/or one or more coding sequences. A vector may be a viral vector, bacteriophage, bacterial artificial chromosome, plasmid, cosmid, or yeast artificial chromosome. A vector may be a DNA or RNA vector. A vector may be a self-replicating extrachromosomal vector, Viral vectors include, but are not limited to, adenovirus vector, adeno-associated virus (AAV) vector, retrovirus vector, or lentivirus vector. A vector may be an adeno-associated virus (AAV) vector. The vector may encode a Cas9 protein or fusion protein and at least one gRNA molecule. 
     Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. For example, any nomenclatures used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization described herein are those that are well known and commonly used in the art. The meaning and scope of the terms should be clear; in the event however of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. 
     2. Fusion Protein 
     Provided herein is a fusion protein. The fusion protein may activate transcription of a target gene. The fusion protein may increase transcription or expression of a target gene by at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 90%, at least about 95%, at least about 200%, at least about 300%, at least about 400%, or at least about 500% relative to a control. The fusion protein may increase transcription or expression of a target gene by less than 1,000,000-fold, less than 500,000-fold, less than 100,000-fold, less than 10,000-fold, less than 1,000-fold, less than 100-fold, 10-fold, less than 5-fold, less than 4-fold, less than 3-fold, or less than 2-fold relative to a control. The control may be, for example, transcription or expression of the target gene in a cell in which the fusion protein was not introduced. 
     Activation of transcription or expression of a target gene may include an increase in the level of mRNA expression from the target gene, relative to a control. The control may be, for example, the level of mRNA expression of the target gene in a cell lacking the fusion protein. The mRNA expression level from the target gene may be increased at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 100-fold, at least 500-fold, at least 1,000-fold, at least 5,000-fold, at least 10,000-fold, at least 15,000-fold, at least 20,000-fold, at least 25,000-fold, at least 30,000-fold, at least 50,000-fold, or at least 100,000-fold relative to a control. The mRNA expression level from the target gene may be increased less than 1,000,000-fold, less than 500,000-fold, less than 100,000-fold, less than 50,000-fold, less than 40,000-fold, less than 30,000-fold, less than 25,000-fold, less than 20,000-fold, less than 15,000-fold, less than 10,000-fold, less than 5,000-fold, less than 1,000-fold, less than 500-fold, less than 100-fold, less than 50-fold, or less than 10-fold relative to a control. The mRNA expression level from the target gene may be increased 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to 500-fold, 2-fold to 1,000-fold, 2-fold to 5,000-fold, 2-fold to 10,000-fold, 2-fold to 15,000-fold, 2-fold to 20,000-fold, 2-fold to 25,000-fold, 2-fold to 30,000-fold, 2-fold to 50,000-fold, 2-fold to 100,000-fold, 2-fold to 500,000-fold, 2-fold to 1,000,000-fold, 10-fold to 50-fold, 10-fold to 100-fold, 10-fold to 500-fold, 10-fold to 1,000-fold, 10-fold to 5,000-fold, 10-fold to 10,000-fold, 10-fold to 15,000-fold, 10-fold to 20,000-fold, 10-fold to 25,000-fold, 10-fold to 30,000-fold, 10-fold to 50,000-fold, 10-fold to 100,000-fold, 10-fold to 500,000-fold, 10-fold to 1,000,000-fold, 100-fold to 500-fold, 100-fold to 1,000-fold, 100-fold to 5,000-fold, 100-fold to 10,000-fold, 100-fold to 15,000-fold, 100-fold to 20,000-fold, 100-fold to 25,000-fold, 100-fold to 30,000-fold, 100-fold to 50,000-fold, 100-fold to 100,000-fold, 100-fold to 500,000-fold, 100-fold to 1,000,000-fold, 1,000-fold to 5,000-fold, 1,000-fold to 10,000-fold, 1,000-fold to 15,000-fold, 1,000-fold to 20,000-fold, 1,000-fold to 25,000-fold, 1,000-fold to 30,000-fold, 1,000-fold to 50,000-fold, 1,000-fold to 100,000-fold, 1,000-fold to 500,000-fold, or 1,000-fold to 1,000,000-fold relative to a control. 
     Activation of transcription or expression of a target gene may include an increase in the level of protein expressed from the target gene, relative to a control. The control may be, for example, the level of protein expressed from the target gene in a cell lacking the fusion protein. The level of protein expression from the target gene may be increased at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 20-fold, at least 30-fold, at least 40-fold, at least 50-fold, at least 100-fold, at least 500-fold, at least 1,000-fold, at least 5,000-fold, at least 10,000-fold, at least 15,000-fold, at least 20,000-fold, at least 25,000-fold, at least 30,000-fold, at least 50,000-fold, or at least 100,000-fold relative to a control. The level of protein expression from the target gene may be increased less than 1,000,000-fold, less than 500,000-fold, less than 100,000-fold, less than 50,000-fold, less than 40,000-fold, less than 30,000-fold, less than 25,000-fold, less than 20,000-fold, less than 15,000-fold, less than 10,000-fold, less than 5,000-fold, less than 1,000-fold, less than 500-fold, less than 100-fold, less than 50-fold, or less than 10-fold relative to a control. The level of protein expression from the target gene may be increased 2-fold to 50-fold, 2-fold to 100-fold, 2-fold to 500-fold, 2-fold to 1,000-fold, 2-fold to 5,000-fold, 2-fold to 10,000-fold, 2-fold to 15,000-fold, 2-fold to 20,000-fold, 2-fold to 25,000-fold, 2-fold to 30,000-fold, 2-fold to 50,000-fold, 2-fold to 100,000-fold, 2-fold to 500,000-fold, 2-fold to 1,000,000-fold, 10-fold to 50-fold, 10-fold to 100-fold, 10-fold to 500-fold, 10-fold to 1,000-fold, 10-fold to 5,000-fold, 10-fold to 10,000-fold, 10-fold to 15,000-fold, 10-fold to 20,000-fold, 10-fold to 25,000-fold, 10-fold to 30,000-fold, 10-fold to 50,000-fold, 10-fold to 100,000-fold, 10-fold to 500,000-fold, 10-fold to 1,000,000-fold, 100-fold to 500-fold, 100-fold to 1,000-fold, 100-fold to 5,000-fold, 100-fold to 10,000-fold, 100-fold to 15,000-fold, 100-fold to 20,000-fold, 100-fold to 25,000-fold, 100-fold to 30,000-fold, 100-fold to 50,000-fold, 100-fold to 100,000-fold, 100-fold to 500,000-fold, 100-fold to 1,000,000-fold, 1,000-fold to 5,000-fold, 1,000-fold to 10,000-fold, 1,000-fold to 15,000-fold, 1,000-fold to 20,000-fold, 1,000-fold to 25,000-fold, 1,000-fold to 30,000-fold, 1,000-fold to 50,000-fold, 1,000-fold to 100,000-fold, 1,000-fold to 500,000-fold, or 1,000-fold to 1,000,000-fold relative to a control. 
     The fusion protein comprises at least two heterologous polypeptide domains. The first polypeptide domain comprises a DNA binding protein. The first polypeptide domain is fused to at least one second polypeptide domain. The second polypeptide domain comprises a modulator of chromatin structure. In some embodiments, the fusion protein further includes at least one third polypeptide domain. The third polypeptide domain comprises a transcriptional activator domain. 
     The linkage to the first polypeptide domain, to the second polypeptide domain, and/or to the third polypeptide domain can be through reversible or irreversible covalent linkage or through a non-covalent linkage, as long as the linker does not interfere with the function of the first, second, or third polypeptide domain(s). For example, a DNA binding protein can be linked to a second polypeptide domain as part of a fusion protein. As another example, they can be linked through reversible non-covalent interactions such as avidin (or streptavidin)-biotin interaction, histidine-divalent metal ion interaction (such as, Ni, Co, Cu, Fe), interactions between multimerization (such as, dimerization) domains, or glutathione 5-transferase (GST)-glutathione interaction. As yet another example, they can be linked covalently but reversibly with linkers such as dibromomaleimide (DBM) or amino-thiol conjugation. 
     In some embodiments, the fusion protein further includes at least one linker. A linker may be included anywhere in the polypeptide sequence of the fusion protein, for example, between the first and second domains, between the first and third domains, and/or between the second and third domains. A linker may be of any length and design to promote or restrict the mobility of components in the fusion protein. A linker may comprise any amino acid sequence of about 2 to about 100, about 5 to about 80, about 10 to about 60, or about 20 to about 50 amino acids. A linker may comprise an amino acid sequence of at least about 2, 3, 4, 5, 10, 15, 20, 25, or 30 amino acids. A linker may comprise an amino acid sequence of less than about 100, 90, 80, 70, 60, 50, or 40 amino acids. A linker may include sequential or tandem repeats of an amino acid sequence that is 2 to 20 amino acids in length. Linkers may include, for example, a GS linker (Gly-Gly-Gly-Gly-Ser) n , wherein n is an integer between 0 and 10 (SEQ ID NO: 55). In a GS linker, n can be adjusted to optimize the linker length and achieve appropriate separation of the functional domains. Other examples of linkers may include, for example, Gly-Gly-Gly-Gly-Gly (SEQ ID NO: 56), Gly-Gly-Ala-Gly-Gly (SEQ ID NO: 57), Gly/Ser rich linkers such as Gly-Gly-Gly-Gly-Ser-Ser-Ser (SEQ ID NO: 58), or Gly/Ala rich linkers such as Gly-Gly-Gly-Gly-Ala-Ala-Ala (SEQ ID NO: 59). 
     In some embodiments, the fusion protein includes a nuclear localization sequence (NLS). Nuclear localization sequences are known in the art. Nuclear localization sequences include, for example, SV40 NLS (Pro-Lys-Lys-Lys-Arg-Lys-Val: SEQ ID NO: 60). 
     a. First Domain: DNA Binding Protein 
     The first polypeptide domain of the fusion protein comprises a DNA binding protein. The DNA binding protein may be a zinc finger protein, a transcription activator-like effector (TALE), or a Cas protein. The DNA binding protein targets the fusion protein for binding to a specific DNA sequence. 
     Alternative to a DNA binding protein, or in addition to, the fusion protein may include an aptamer. Aptamers are polynucleotides or polypeptides that specifically recognize and bind to a specific target molecule, such as to a DNA sequence. 
     i) Zinc Finger Protein 
     The DNA binding protein may comprise a zinc finger protein. A zinc finger protein is a protein that includes one or more zinc finger domains. Zinc finger domains are relatively small protein motifs that contain multiple finger-like protrusions that make tandem contacts with their target molecule such as a DNA target molecule. A zinc finger domain may bind one or more zinc ions or other metal ion such as iron, or in some cases a zinc finger domain forms salt bridges to stabilize the finger-like folds. The zinc binding portion of a zinc finger protein may include one or more cysteine residues and/or one or more histidine residues to coordinate the zinc or other metal ion. A zinc finger protein recognizes and binds to a particular DNA sequence via the zinc finger domain. 
     ii) TALE 
     The DNA binding protein may comprise a transcription activator-like effector (TALE). A TALE is another type of protein that recognizes and binds to a particular DNA sequence. The DNA-binding domain of a TALE includes an array of tandem 33-35 amino acid repeats, also known as RVD modules. Each RVD module specifically recognizes a single base pair of DNA. RVD modules may be arranged in any order to assemble an array that recognizes a defined DNA sequence. The binding specificity of a TALE DNA-binding domain is determined by the RVD array followed by a single truncated repeat of, for example, 20 amino acids. A TALE DNA-binding domain may have an array of 12 to 27 RVD modules, each RVD module recognizing a single base pair of DNA. Specific RVDs have been identified that recognize each of the four possible DNA nucleotides (A, T, C, and G). Because the TALE DNA-binding domains are modular, repeats that recognize the four different DNA nucleotides may be linked together to recognize any particular DNA sequence. These targeted DNA-binding domains may then be combined with catalytic domains to create functional enzymes, including artificial transcription factors and/or nucleases. In some embodiments, the TALE specifically binds to a target sequence associated with a target gene. 
     iii) Cas Protein 
     The DNA binding protein may include a Cas protein or a mutated Cas protein. “Clustered Regularly interspaced Short Palindromic Repeats” and “CRISPRs”, as used interchangeably herein, refers to loci containing multiple short direct repeats that are found in the genomes of approximately 40% of sequenced bacteria and 90% of sequenced archaea. The CRISPR system is a microbial nuclease system involved in defense against invading phages and plasmids that provides a form of acquired immunity. The CRISPR loci in microbial hosts contain a combination of CRISPR-associated (Cas) genes as well as non-coding RNA elements capable of programming the specificity of the CRISPR-mediated nucleic acid cleavage. Short segments of foreign DNA, called spacers, are incorporated into the genome between CRISPR repeats, and serve as a “memory” of past exposures. Cas proteins include, for example, Cas9, Cas12, and Cas12a. In some embodiments, the Cas protein is a Cas9 protein, Cas9 forms a complex with the 3′ end of the sgRNA (which may be referred interchangeably herein as “gRNA”), and the protein-RNA pair recognizes its genomic target by complementary base pairing between the 5′ end of the sgRNA sequence and a predefined 20 bp DNA sequence, known as the protospacer. This complex is directed to homologous loci of pathogen DNA via regions encoded within the crRNA, i.e., the protospacers, and protospacer-adjacent motifs (PAMs) within the pathogen genome. The non-coding CRISPR array is transcribed and cleaved within direct repeats into short crRNAs containing individual spacer sequences, which direct Cas nucleases to the target site (protospacer). By simply exchanging the 20 by recognition sequence of the expressed sgRNA, the Cas9 nuclease can be directed to new genomic targets. CRISPR spacers are used to recognize and silence exogenous genetic elements in a manner analogous to RNAi in eukaryotic organisms. 
     Three classes of CRISPR systems (Types I, II, and III effector systems) are known. The Cas protein may be from any of the Types I, II, and III effector systems. The Type II effector system carries out targeted DNA double-strand break in four sequential steps, using a single effector enzyme, Cas9, to cleave dsDNA. Compared to the Type I and Type ill effector systems, which require multiple distinct effectors acting as a complex, the Type II effector system may function in alternative contexts such as eukaryotic cells. The Type II effector system consists of a long pre-crRNA, which is transcribed from the spacer-containing CRISPR locus, the Cas9 protein, and a tracrRNA, which is involved in pre-crRNA processing. The tracrRNAs hybridize to the repeat regions separating the spacers of the pre-crRNA, thus initiating dsRNA cleavage by endogenous RNase III. This cleavage is followed by a second cleavage event within each spacer by Cas9, producing mature crRNAs that remain associated with the tracrRNA and Cas9, forming a Cas9:crRNA-tracrRNA complex. 
     The Cas9:crRNA-tracrRNA complex unwinds the DNA duplex and searches for sequences matching the crRNA to cleave. Target recognition occurs upon detection of complementarily between a “protospacer” sequence in the target DNA and the remaining spacer sequence in the crRNA. Cas9 mediates cleavage of target DNA if a correct protospacer-adjacent motif (PAM) is also present at the 3′ end of the protospacer. For protospacer targeting, the sequence must be immediately followed by the protospacer-adjacent motif (PAM), a short sequence recognized by the Cas9 nuclease that is required for DNA cleavage. Different Type 11 systems have differing PAM requirements. 
     An engineered form of the Type H effector system of  Streptococcus pyogenes  was shown to function in human cells for genome engineering. In this system, the Cas9 protein was directed to genomic target sites by a synthetically reconstituted “guide RNA” (“gRNA”, also used interchangeably herein as a chimeric single guide RNA (“sgRNA”)), which is a crRNA-tracrRNA fusion that obviates the need for RNase III and crRNA processing in general. Provided herein are CRISPR/Cas9-based engineered systems for use in gene editing and treating genetic diseases. The CRISPR/Cas9-based engineered systems can be designed to target any gene, including genes involved in, for example, a genetic disease, aging, tissue regeneration, or wound healing. The CRISPR/Cas9-based gene editing system can include a Cas9 protein or a Cas9 fusion protein. 
     Cas9 protein is an endonuclease that cleaves nucleic acid and is encoded by the CRISPR loci and is involved in the Type II CRISPR system. The Cas9 protein can be from any bacterial or archaea species, including, but not limited to,  Streptococcus pyogenes, Staphylococcus aureus  ( S. aureus ),  Acidovorax avenae, Actinobacillus pleuropneurnoniae, Actinobacillus succinogenes, Actinobacillus suis, Actinomyces  sp., cycliphilus  denitrificans, Aminomonas paucivorans, Bacillus cereus, Bacillus smithii, Bacillus thuringiensis, Bacteroides  sp.,  Blastopirellula marina , Bradyrhizobiurn sp., Brevibacilius  laterosporus, Campylobacter coli, Campylobacter jejuni, Campylobacter lari , Candidatus Puniceispirillum,  Clostridium cellulolyticum, Clostridium perfringens, Corynebacterium accolens, Corynebacteriurn diphtheria, Corynebacterium matruchotii, Dinomseobacter shibae, Eubacterium dolichum , gamma proteobacterium,  Gluconacetobacter diazotrophicus, Haemophilus parainfluenzae, Haemophilus sputorum, Helicobacter canadensis, Helicobacter cinaedi, Helicobacter musteiae, ilyobacter polytropus, Kingella kingae, Lactobacillus crispatus, Listeria ivanovii, Listeria monocytogenes, Listeriaceae bacterium, Methylocystis  sp.,  Methylosinus trichosporium, Mobiluncus mulieris, Neisseria bacilliformis, Neisseria cinerea, Neisseria flavescens, Neisseria lactamica, Neisseria  sp.,  Neisseria wadsworthii, Nitrosomonas  sp.,  Parvibaculum lavamentivorans, Pasteurella multocida, Phascolarctobacterium succinatutens, Ralstonia syzygii, Rhodopseudomonas palustris, Rhodovulum  sp.,  Simonsiella muelleri, Sphingomonas  sp.,  Sporolactobacillus vineae, Staphylococcus lugdunensis, Streptococcus  sp.,  Subdoligranulum  sp.,  Tistrella Treponema  sp., or  Verminephrobacter eiseniae . In certain embodiments, the Cas9 molecule is a  Streptococcus pyogenes  Cas9 molecule (also referred herein as “SpCas9”). SpCas9 may comprise an amino acid sequence of SEQ ID NO: 18. In certain embodiments, the Cas9 molecule is a  Staphylococcus aureus  Cas9 molecule (also referred herein as “SaCas9”). SaCas9 may comprise an amino acid sequence of SEQ ID NO: 19. 
     A Cas9 molecule or a Cas9 fusion protein can interact with one or more gRNA molecule(s) and, in concert with the gRNA molecule(s), can localize to a site which comprises a target domain, and in certain embodiments, a PAM sequence. The Cas9 protein forms a complex with the 3′ end of a gRNA. The ability of a Cas9 molecule or a Cas9 fusion protein to recognize a PAM sequence can be determined, for example, by using a transformation assay as known in the art. 
     The specificity of the CRISPR-based system may depend on two factors: the target sequence and the protospacer-adjacent motif (PAM). The target sequence is located on the 5′ end of the gRNA and is designed to bond with base pairs on the host DNA at the correct DNA sequence known as the protospacer. By simply exchanging the recognition sequence of the gRNA, the Cas9 protein can be directed to new genomic targets. The PAM sequence is located on the DNA to be altered and is recognized by a Cas9 protein. PAM recognition sequences of the Cas9 protein can be species specific. 
     In certain embodiments, the ability of a Cas9 molecule or a Cas9 fusion protein to interact with and cleave a target nucleic acid is PAM sequence dependent. A PAM sequence is a sequence in the target nucleic acid. In certain embodiments, cleavage of the target nucleic acid occurs upstream from the PAM sequence. Cas9 molecules from different bacterial species can recognize different sequence motifs (for example, PAM sequences). A Cas9 molecule of  S. pyogenes  may recognize the PAM sequence of NRG (5′-NRG-3′, where R is any nucleotide residue, and in some embodiments, R is either A or G, SEQ ID NO: 1). In certain embodiments, a Cas9 molecule of  S. pyogenes  may naturally prefer and recognize the sequence motif NGG (SEQ ID NO: 2) and directs cleavage of a target nucleic acid sequence 1 to 10, for example, 3 to 5, bp upstream from that sequence. In some embodiments, a Cas9 molecule of  S. pyogenes  accepts other PAM sequences, such as NAG (SEQ ID NO: 3) in engineered systems (Hsu et al., Nature Biotechnology 2013 doi:10.1038/nbt.2647). In certain embodiments, a Cas9 molecule of  S. thermophilus  recognizes the sequence motif NGGNG (SEQ ID NO: 4) and/or NNAGAAW (W=A or T) (SEQ ID NO: 5) and directs cleavage of a target nucleic add sequence 1 to 10, for example, 3 to 5, bp upstream from these sequences. In certain embodiments, a Cas9 molecule of  S. mutans  recognizes the sequence motif NGG (SEQ ID NO: 2) and/or NAAR (R=A or G) (SEQ ID NO: 6) and directs cleavage of a target nucleic acid sequence 1 to 10, for example, 3 to 5 bp, upstream from this sequence. In certain embodiments, a Cas9 molecule of  S. aureus  recognizes the sequence motif NNGRR (R=A or G) (SEQ ID NO: 7) and directs cleavage of a target nucleic acid sequence 1 to 10, for example, 3 to 5, bp upstream from that sequence. In certain embodiments, a Cas9 molecule of  S. aureus  recognizes the sequence motif NNGRRN (R=A or G) (SEQ ID NO: 8) and directs cleavage of a target nucleic acid sequence 1 to 10, for example, 3 to 5, bp upstream from that sequence. In certain embodiments, a Cas9 molecule of  S. aureus  recognizes the sequence motif NNGRRT (R=A or G) (SEQ ID NO: 9) and directs cleavage of a target nucleic acid sequence 1 to 10, for example, 3 to 5, bp upstream from that sequence. In certain embodiments, a Cas9 molecule of  S. aureus  recognizes the sequence motif NNGRRV (R=A or G; V=A or C or G) (SEQ ID NO: 10) and directs cleavage of a target nucleic acid sequence 1 to 10, for example, 3 to 5, bp upstream from that sequence. A Cas9 molecule derived from  Neisseria meningitidis  (NmCas9) normally has a native PAM of NNNNGATT (SEQ ID NO: 11), but may have activity across a variety of PAMs, including a highly degenerate NNNNGNNN PAM (SEQ ID NO: 12) (Esvelt et al. Nature Methods 2013 doi:10.1038/nmeth.2881). In the aforementioned embodiments, N can be any nucleotide residue, for example, any of A, G, C, or T. Cas9 molecules can be engineered to alter the PAM specificity of the Cas9 molecule. 
     In some embodiments, the Cas9 protein recognizes a PAM sequence NGG (SEQ ID NO: 2) or NGA (SEQ ID NO: 13) or NNNRRT (R=A or G; SEQ ID NO: 14) or ATTCCT (SEQ ID NO: 15) or NGAN (SEQ ID NO: 16) or NGNG (SEQ ID NO: 17). In some embodiments, the Cas9 protein is a Cas9 protein of  S. aureus  and recognizes the sequence motif NNGRR (R=A or G; SEQ ID NO: 7), NNGRRN (R=A or G; SEQ ID NO: 8), NNGRRT (R=A or G; SEQ ID NO: 9), or NNGRRV (R=A or G; SEQ ID NO: 10). In the aforementioned embodiments, N can be any nucleotide residue, for example, any of A, G, C, or T. 
     In some embodiments, the at least one Cas9 molecule is a mutant Cas9 molecule. The Cas9 protein can be mutated so that the nuclease activity is inactivated. An inactivated Cas9 protein (“iCas9”, also referred to as “dCas9”) with no endonuclease activity has been targeted to genes in bacteria, yeast, and human cells by gRNAs to silence gene expression through steric hindrance. Exemplary mutations with reference to the  S. pyogenes  Cas9 sequence to inactivate the nuclease activity include: D10A, E762A, H840A, N854A, N863A and/or D986A, A  S. pyogenes  Cas9 protein with the D10A mutation may comprise an amino acid sequence of SEQ ID NO: 20. In some embodiments, the dCas9 protein comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 20, or any range between any two of these values. A  S. pyogenes  Cas9 protein with D10A and H849A mutations may comprise an amino acid sequence of SEQ ID NO: 21. In some embodiments, the dCas9 protein comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 21, or any range between any two of these values. Exemplary mutations with reference to the  S. aureus  Cas9 sequence to inactivate the nuclease activity include Di OA and N580A. In certain embodiments, the mutant  S. aureus  Cas9 molecule comprises a D10A mutation. The nucleotide sequence encoding this mutant  S. aureus  Cas9 is set forth in SEQ ID NO: 22. In some embodiments, the dCas9 protein comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 22, or any range between any two of these values. In certain embodiments, the mutant  S. aureus  Cas9 molecule comprises a N580A mutation. The nucleotide sequence encoding this mutant  S. aureus  Cas9 molecule is set forth in SEQ ID NO: 23. In some embodiments, the dCas9 protein comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 23, or any range between any two of these values. 
     In some embodiments, the Cas9 protein is a VQR variant. The VQR variant of Cas9 is a mutant with a different PAM recognition, as detailed in Kleinstiver, et al. (Nature 2015, 523, 481-485, incorporated herein by reference). 
     A polynucleotide encoding a Cas9 molecule can be a synthetic polynucleotide. For example, the synthetic polynucleotide can be chemically modified. The synthetic polynucleotide can be codon optimized, for example, at least one non-common codon or less-common codon has been replaced by a common codon. For example, the synthetic polynucleotide can direct the synthesis of an optimized messenger mRNA, for example, optimized for expression in a mammalian expression system, as described herein. An exemplary codon optimized nucleic acid sequence encoding a Cas9 molecule of  S. pyogenes  is set forth in SEQ ID NO: 24. Exemplary codon optimized nucleic acid sequences encoding a Cas9 molecule of  S. aureus , and optionally containing nuclear localization sequences (NLSs), are set forth in SEQ ID NOs: 25-31. Another exemplary codon optimized nucleic acid sequence encoding a Cas9 molecule of  S. aureus  comprises the nucleotides 1293-4451 of SEQ ID NO: 32. 
     b. Second Domain: Modulator of Chromatin Remodeling 
     The second polypeptide domain comprises a modulator of chromatin structure. The modulator of chromatin structure may also be referred to as a chromatin remodeling protein. The modulator of chromatin structure may have an activity selected from destabilizing histone-DNA interactions, destabilizing nucleosomes, promoting movement of DNA relative to histories (for example, sliding DNA along histones and/or translocating DNA along histones and/or changing the position of a nucleosome, relative to an associated DNA strand), ejecting nucleosomes from a region of DNA, or ejecting histones from the nucleosome, or a combination thereof. In some embodiments, the modulator of chromatin structure comprises a nucleosome rearranging protein. In some embodiments, the modulator of chromatin structure creates nucleosome-depleted region(s) in a gene or genome. For example, the modulator of chromatin structure may comprise the SS18 subunit of the BAF chromatin remodeling complex. The BAF chromatin remodeling complex may also be referred to as the SWItch/Sucrose Non-Fermentable (SWI/SNF) chromatin remodeling complex. SWI/SNF is a subfamily of ATP-dependent chromatin remodeling complexes. The remodeling complex may be composed of several proteins that are products of the SWI and SNF genes, such as SWI1, SWI2/SNF2, SWI3, SWI5, and SWI6. The remodeling complex has a DNA-stimulated ATPase activity that can destabilize histone-DNA interactions in reconstituted nucleosomes in an ATP-dependent manner. The SWI/SNF subfamily may provide nucleosome rearrangement, such as ejection and/or sliding. The movement of nucleosomes may provide easier access to the chromatin, allowing gene expression to be activated or repressed. In some embodiments, the modulator of chromatin structure comprises CHD1 or CHD8 or a variant thereof. In some embodiments, the modulator of chromatin structure comprises the BAF chromatin remodeling complex or a functional subunit thereof or a variant thereof. In some embodiments, the modulator of chromatin structure is a protein that recruits the BAF complex or subunits thereof. The modulator of chromatin structure may comprise the SS18 subunit or a variant thereof. SS18 is a member of the human SWI/SNF chromatin remodeling complex and is involved in chromosomal translocation. 5518 may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 37, encoded by the polynucleotide of SEQ ID NO: 38. In some embodiments, SS18 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 37, or any range between any two of these values. In some embodiments, the modulator of chromatin structure comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 18, 17, 18, 19, or 20 amino acid substitutions relative to wild-type SS18 protein. In some embodiments, the modulator of chromatin structure comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid substitutions relative to SEQ ID NO: 37, In some embodiments, 5518 comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 38, or any range between any two of these values. The modulator of chromatin structure may be from a mammal, such as a mouse or a human, or from another species. In some embodiments, the modulator of chromatin structure is from a mammal. In some embodiments, the modulator of chromatin structure is from a mouse. In some embodiments, the modulator of chromatin structure is from humans. dCas9-SS18 may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 88 or 90, encoded by the polynucleotide of SEQ ID NO: 87 or 89, respectively. In some embodiments, dCas9-SS18 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 88 or 90, or any range between any two of these values. In some embodiments, dCas9-SS18 comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 87 or 89, or any range between any two of these values. 
     In some embodiments, the modulator of chromatin structure does not have an activity selected from acetyltransferase activity, methyltransferase activity, deacetylase activity, or demethylase activity, or a combination thereof. In some embodiments, the modulator of chromatin structure does not have an activity selected from acetyltransferase activity, methyltransferase activity, deacetylase activity, demethylase activity, covalent histone modification activity, binding to or recruitment of a transcription factor such as a transcription activation factor, or a combination thereof. 
     The fusion protein comprises one or more second polypeptide domain(s). For example, the fusion protein may include one, two, three, four, or five second polypeptide domains. The first polypeptide domain and the second polypeptide domain(s) may be operably linked. The second polypeptide domain may be at the C-terminal end of the first polypeptide domain, or at the N-terminal end of the first polypeptide domain, or a combination thereof. The fusion protein may include one second polypeptide domain. The fusion protein may include two of the second polypeptide domains. For example, the fusion protein may include a second polypeptide domain at the N-terminal end of the first polypeptide domain as well as a second polypeptide domain at the C-terminal end of the first polypeptide domain. In other embodiments, the fusion protein may include a single first polypeptide domain and more than one (for example, two or three or four) second polypeptide domains in tandem. Each second polypeptide domain may be the same or different. In some embodiments, the fusion protein comprises SS18 fused to the N-terminal end of dCas9 protein. In some embodiments, the fusion protein comprises SS18 fused to the C-terminal end of dCas9 protein. 
     c. Third Domain: Transcriptional Activator Domain 
     In some embodiments, the fusion protein further includes one or more third polypeptide domain(s). The third polypeptide domain can have transcription activation activity, for example, a transactivation domain. The transcriptional activator domain may have an activity selected from acetyltransferase activity, methyltransferase activity, deacetylase activity, demethylase activity, or a combination thereof. The transcriptional activator domain may have an activity selected from acetyltransferase activity, methyltransferase activity, deacetylase activity, demethylase activity, covalent histone modification activity, binding to or recruitment of a transcription factor such as a transcription activation factor, or a combination thereof. The transcriptional activator domains may include, for example, a VP16 protein, multiple VP16 proteins such as a VP48 domain or VP64 domain, p65 domain of NF kappa B transcription activator activity, activation domain of HSF1, TET1, VPR, VPH, Rta, p300, or p300 core (p300c), or a combination thereof. The third polypeptide domain may be from a mammal, such as a mouse or a human, or from another species. In some embodiments, the third polypeptide domain is from a mammal. In some embodiments, the transcription activator domain is from mouse. In some embodiments, the transcription activator domain is from human. 
     The fusion protein may include, for example, one, two, three, four, or five third polypeptide domains. The first polypeptide domain and the second polypeptide domain(s) and the third polypeptide domain(s) may be operably linked. The third polypeptide domain may be at the C-terminal end of the first polypeptide domain, or at the N-terminal end of the first polypeptide domain, or a combination thereof. The fusion protein may include one third polypeptide domain. The fusion protein may include two of the third polypeptide domains. For example, the fusion protein may include a third polypeptide domain at the N-terminal end of the first polypeptide domain as well as a third polypeptide domain at the C-terminal end of the first polypeptide domain. In other embodiments, the fusion protein may include a single first polypeptide domain and more than one (for example, two or three or four) third polypeptide domains in tandem. Each third polypeptide domain may be the same or different. 
     In some embodiments, the transcriptional activator domain comprises p300. p300 may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 33, or of SEQ ID NO: 34 (p300c). In some embodiments, p300 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 33 or SEQ ID NO: 34, or any range between any two of these values. dCas9-p300c may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 80 or 82, encoded by the polynucleotide of SEQ ID NO: 79 or 81, respectively. In some embodiments, dCas9-p300c comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 80 or 82, or any range between any two of these values. In some embodiments, dCas9-p300c comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 79 or 81, or any range between any two of these values. P300c-dCas9 may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 84 or 86, encoded by the polynucleotide of SEQ ID NO: 83 or 85, respectively. In some embodiments, p300c-dCas9 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 84 or 86, or any range between any two of these values. In some embodiments, p300c-dCas9 comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 83 or 85, or any range between any two of these values. In some embodiments, the fusion protein comprises p300 fused to the N-terminal end of dCas9 protein. In some embodiments, the fusion protein comprises p300 fused to the C-terminal end of dCas9 protein. 
     In some embodiments, the fusion protein comprises TET1. TET1, also known as Tet1CD (Ten-eleven translocation methylcytosine dioxygenase 1), may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 93, encoded by the polynucleotide of SEQ ID NO: 94. In some embodiments, TET1 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 93, or any range between any two of these values. In some embodiments, TET1 comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 94, or any range between any two of these values. In some embodiments, the fusion protein comprises TET1 fused to the N-terminal end of dCas9 protein. In some embodiments, the fusion protein comprises TET1 fused to the C-terminal end of dCas9 protein. 
     In some embodiments, the fusion protein comprises VP64. VP64 may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 91, encoded by the polynucleotide of SEQ ID NO: 92. In some embodiments, VP64 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 91, or any range between any two of these values. In some embodiments, VP64 comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 92, or any range between any two of these values. In some embodiments, the fusion protein comprises VP64-dCas9-VP64. VP64-dCas9-VP64 may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 35, encoded by the polynucleotide of SEQ ID NO: 36. In some embodiments, VP64-dCas9-VP64 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 35, or any range between any two of these values. In some embodiments, VP64-dCas9-VP64 comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 36, or any range between any two of these values. In some embodiments, the fusion protein comprises VP64 fused to the N-terminal end of dCas9 protein. In some embodiments, the fusion protein comprises VP64 fused to the C-terminal end of dCas9 protein. 
     In some embodiments, the transcriptional activator domain comprises VPH, which is a polypeptide comprising VP64, mouse p65, and HSF1. VPH may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 39, encoded by the polynucleotide of SEQ ID NO: 40. In some embodiments, VPH comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 39, or any range between any two of these values. In some embodiments, VPH comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 40, or any range between any two of these values, dCas9-VPH may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 72 or 74, encoded by the polynucleotide of SEQ ID NO: 71 or 73, respectively. In some embodiments, dCas9-VPH comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 72 or 74, or any range between any two of these values. In some embodiments, dCas9-VPH comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 71 or 73, or any range between any two of these values. VPH-dCas9 may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 68 or 70, encoded by the polynucleotide of SEQ ID NO: 67 or 69, respectively. In some embodiments, VPH-dCas9 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 68 or 70, or any range between any two of these values. In some embodiments, VPH-dCas9 comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 67 or 69, or any range between any two of these values. VPH-dCas9-VPH may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 76, encoded by the polynucleotide of SEQ ID NO: 75. In some embodiments, VPH-dCas9-VPH comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 76, or any range between any two of these values. In some embodiments, VPH-dCas9-VPH comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 75, or any range between any two of these values. In some embodiments, the fusion protein comprises VPH fused to the N-terminal end of dCas9 protein. In some embodiments, the fusion protein comprises VPH fused to the C-terminal end of dCas9 protein. 
     In some embodiments, the transcriptional activator domain comprises VPR, which is a polypeptide comprising VP64, human p65, and Rta. VPR may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 41, encoded by the polynucleotide of SEQ ID NO: 42. In some embodiments, VPR comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 41, or any range between any two of these values. In some embodiments, VPR comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 42, or any range between any two of these values. dCas9-VPR may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 78, encoded by the polynucleotide of SEQ ID NO: 77. In some embodiments, dCas9-VPR comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 78, or any range between any two of these values. In some embodiments, dCas9-VPR comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 77, or any range between any two of these values. In some embodiments, the fusion protein comprises VPR fused to the N-terminal end of dCas9 protein. In some embodiments, the fusion protein comprises VPR fused to the C-terminal end of dCas9 protein. 
     In some embodiments, the fusion protein comprises dCas9 protein with VPR fused to its C-terminal end and SS18 fused to its N-terminal end. In some embodiments, the fusion protein comprises dCas9 protein with VPR fused to its N-terminal end and SS18 fused to its C-terminal end. For example, the fusion protein may comprise VPR-dCas9-SS18 or SS18-dCas9-VPR (N-terminal end to C-terminal end) or variants thereof. In some embodiments, the fusion protein comprises dCas9 protein with p300 or p300-core fused to its C-terminal end and SS18 fused to its N-terminal end. In some embodiments, the fusion protein comprises dCas9 protein with p300 or p300-core fused to its N-terminal end and SS18 fused to its C-terminal end. For example, the fusion protein may comprise p300-dCas9-SS18 or SS18-dCas9-p300 (N-terminal end to C-terminal end) or variants thereof. In some embodiments, the fusion protein comprises dCas9 protein with VP64 fused to its C-terminal end and SS18 fused to its N-terminal end. In some embodiments, the fusion protein comprises dCas9 protein with VP64 fused to its N-terminal end and SS18 fused to its C-terminal end. For example, the fusion protein may comprise VP64-dCas9-SS18 or SS18-dCas9-VP64 (N-terminal end to C-terminal end) or variants thereof. In some embodiments, the fusion protein comprises dCas9 protein with VPH fused to its C-terminal end and SS18 fused to its N-terminal end. In some embodiments, the fusion protein comprises dCas9 protein with VPH fused to its N-terminal end and SS18 fused to its C-terminal end. For example, the fusion protein may comprise VPH-dCas9-SS18 or SS18-dCas9-VPH (N-terminal end to C-terminal end) or variants thereof. VPH-dCas9-SS18 may comprise a polypeptide having the amino acid sequence of SEQ ID NO: 64 or 66, encoded by the polynucleotide of SEQ ID NO: 63 or 65, respectively. In some embodiments, VPH-dCas9-SS18 comprises a polypeptide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 64 or 66, or any range between any two of these values. In some embodiments, VPH-dCas9-SS18 comprises a polypeptide encoded by a polynucleotide having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 63 or 65, or any range between any two of these values. 
     3. DNA Targeting System 
     Provided herein are DNA Targeting Systems. The DNA Targeting System may be used to activate transcription or increase expression of a gene. The DNA Targeting System includes at least one fusion protein as detailed herein. In embodiments wherein the DNA binding protein of the fusion protein comprises a Cas protein such as Cas9, the DNA Targeting System may further include at least one gRNA. 
     a. Guide RNA (gRNA) 
     The at least one gRNA molecule can bind and recognize a target region. The gRNA provides the targeting of a Cas9 DNA targeting system, which may also be referred to as a CRISPR/Cas9-based gene editing system. The gRNA is a fusion of two noncoding RNAs: a crRNA and a tracrRNA, gRNA mimics the naturally occurring crRNA:tracrRNA duplex involved in the Type II Effector system. This duplex, which may include, for example, a 42-nucleotide crRNA and a 75-nucleotide tracrRNA, acts as a guide for the Cas9 to bind, and in some cases, cleave the target nucleic acid. The gRNA may target any desired DNA sequence by exchanging the sequence encoding a 20 by protospacer which confers targeting specificity through complementary base pairing with the desired DNA target. The “target region” or “target sequence” refers to the region of the target gene to which the CRISPR/Cas9-based gene editing system targets and binds. The portion of the gRNA that targets the target sequence in the genome may be referred to as the “targeting sequence” or “targeting portion” or “targeting domain.” “Protospacer” or “gRNA spacer” may refer to the region of the target gene to which the CRISPR/Cas9-based gene editing system targets and binds; “protospacer” or “gRNA spacer” may also refer to the portion of the gRNA that is complementary to the targeted sequence in the genome. The gRNA may include a gRNA scaffold. A gRNA scaffold facilitates Cas9 binding to the gRNA and may facilitate endonuclease activity. The gRNA scaffold is a polynucleotide sequence that follows the portion of the gRNA corresponding to the sequence that the gRNA targets. Together, the gRNA targeting portion and gRNA scaffold form one polynucleotide. The constant region of the gRNA may include the sequence of SEQ ID NO: 62 (RNA), which is encoded by a sequence comprising SEQ ID NO: 61 (DNA). The CRISPR/Cas9-based gene editing system may include at least one gRNA, wherein the gRNAs target different DNA sequences. The target DNA sequences may be overlapping. The gRNA may comprise at its 5′ end the targeting domain that is sufficiently complementary to the target region to be able to hybridize to, for example, about 10 to about 20 nucleotides of the target region of the target gene, when it is followed by an appropriate Protospacer Adjacent Motif (PAM). The target region or protospacer is followed by a PAM sequence at the 3′ end of the protospacer in the genome. Different Type II systems have differing PAM requirements, as detailed above. 
     The targeting domain of the gRNA does not need to be perfectly complementary to the target region of the target DNA. In some embodiments, the targeting domain of the gRNA is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or at least 99% complementary to (or has 1, 2 or 3 mismatches compared to) the target region over a length of, such as, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides. For example, the DNA-targeting domain of the gRNA may be at least 80% complementary over at least 18 nucleotides of the target region. The target region may be on either strand of the target DNA. 
     As described above, the gRNA molecule comprises a targeting domain (also referred to as targeted or targeting sequence), which is a polynucleotide sequence complementary to the target DNA sequence. The gRNA may comprise a “G” at the 5′ end of the targeting domain or complementary polynucleotide sequence. The CRISPR/Cas9-based gene editing system may use gRNAs of varying sequences and lengths. The targeting domain of a gRNA molecule may comprise at least a 10 base pair, at least a 11 base pair, at least a 12 base pair, at least a 13 base pair, at least a 14 base pair, at least a 15 base pair, at least a 16 base pair, at least a 17 base pair, at least a 18 base pair, at least a 19 base pair, at least a 20 base pair, at least a 21 base pair, at least a 22 base pair, at least a 23 base pair, at least a 24 base pair, at least a 25 base pair, at least a 30 base pair, or at least a 35 base pair complementary polynucleotide sequence of the target DNA sequence followed by a PAM sequence. In certain embodiments, the targeting domain of a gRNA molecule has 19-25 nucleotides in length. In certain embodiments, the targeting domain of a gRNA molecule is 20 nucleotides in length. In certain embodiments, the targeting domain of a gRNA molecule is 21 nucleotides in length. In certain embodiments, the targeting domain of a gRNA molecule is 22 nucleotides in length. In certain embodiments, the targeting domain of a gRNA molecule is 23 nucleotides in length. 
     The number of gRNA molecules that may be included in the CRISPR/Cas9-based gene editing system can be at least 1 gRNA, at least 2 different gRNAs, at least 3 different gRNAs, at least 4 different gRNAs, at least 5 different gRNAs, at least 6 different gRNAs, at least 7 different gRNAs, at least 8 different gRNAs, at least 9 different gRNAs, at least 10 different gRNAs, at least 11 different gRNAs, at least 12 different gRNAs, at least 13 different gRNAs, at least 14 different gRNAs, at least 15 different gRNAs, at least 16 different gRNAs, at least 17 different gRNAs, at least 18 different gRNAs, at least 18 different gRNAs, at least 20 different gRNAs, at least 25 different gRNAs, at least 30 different gRNAs, at least 35 different gRNAs, at least 40 different gRNAs, at least 45 different gRNAs, or at least 50 different gRNAs. The number of gRNA molecules that may be included in the CRISPR/Cas9-based gene editing system can be less than 50 different gRNAs, less than 45 different gRNAs, less than 40 different gRNAs, less than 35 different gRNAs, less than 30 different gRNAs, less than 25 different gRNAs, less than 20 different gRNAs, less than 19 different gRNAs, less than 18 different gRNAs, less than 17 different gRNAs, less than 16 different gRNAs, less than 15 different gRNAs, less than 14 different gRNAs, less than 13 different gRNAs, less than 12 different gRNAs, less than 11 different gRNAs, less than 10 different gRNAs, less than 9 different gRNAs, less than 8 different gRNAs, less than 7 different gRNAs, less than 6 different gRNAs, less than 5 different gRNAs, less than 4 different gRNAs, less than 3 different gRNAs, or less than 2 different gRNAs. The number of gRNAs that may be included in the CRISPR/Cas9-based gene editing system can be between at least 1 gRNA to at least 50 different gRNAs, at least 1 gRNA to at least 45 different gRNAs, at least 1 gRNA to at least 40 different gRNAs, at least 1 gRNA to at least 35 different gRNAs, at least 1 gRNA to at least 30 different gRNAs, at least 1 gRNA to at least 25 different gRNAs, at least 1 gRNA to at least 20 different gRNAs, at least 1 gRNA to at least 16 different gRNAs, at least 1 gRNA to at least 12 different gRNAs, at least 1 gRNA to at least 8 different gRNAs, at least 1 gRNA to at least 4 different gRNAs, at least 4 gRNAs to at least 50 different gRNAs, at least 4 different gRNAs to at least 45 different gRNAs, at least 4 different gRNAs to at least 40 different gRNAs, at least 4 different gRNAs to at least 35 different gRNAs, at least 4 different gRNAs to at least 30 different gRNAs, at least 4 different gRNAs to at least 25 different gRNAs, at least 4 different gRNAs to at least 20 different gRNAs, at least 4 different gRNAs to at least 16 different gRNAs, at least 4 different gRNAs to at least 12 different gRNAs, at least 4 different gRNAs to at least 8 different gRNAs, at least 8 different gRNAs to at least 50 different gRNAs, at least 8 different gRNAs to at least 45 different gRNAs, at least 8 different gRNAs to at least 40 different gRNAs, at least 8 different gRNAs to at least 35 different gRNAs, 8 different gRNAs to at least 30 different gRNAs, at least 8 different gRNAs to at least 25 different gRNAs, 8 different gRNAs to at least 20 different gRNAs, at least 8 different gRNAs to at least 16 different gRNAs, or 8 different gRNAs to at least 12 different gRNAs. 
     The gRNA may target an exon of a gene. The gRNA may target a region of a gene that is non-coding, such as a regulatory region or promoter sequence. The gRNA may target an intron of a gene. In some embodiments, the gRNA corresponds to a polynucleotide sequence selected from SEQ ID NOs: 43-54, a complement of, a truncation thereof, or a variant thereof (TABLE 1). The gRNA may be encoded by or target or bind to or hybridize to a target sequence selected from SEQ ID NOs: 43-48, a complement thereof, a truncation thereof, or a variant thereof. The gRNA may comprise a polynucleotide sequence selected from SEQ ID NOs: 49-54, a complement thereof, a truncation thereof, or a variant thereof. A truncation may be, for example, 1, 2, 3, 4, 5, 8, 7, 8, or 9 nucleotides shorter than the reference sequence. The DNA Targeting System may include one or more gRNAs, each gRNA corresponding to a polynucleotide sequence selected from SEQ ID NOs: 43-54, a complement thereof, a truncation thereof, or a variant thereof, or each gRNA corresponding to a polynucleotide sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% sequence identity to at least one of SEQ ID NOs: 43-54, a complement thereof, a truncation thereof, or a variant thereof. 
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Examples of gRNAs used to activate 
               
               
                 expression of HBG1/2. 
               
            
           
           
               
               
               
            
               
                 Name 
                 gRNA target/ 
                 gRNA 
               
               
                   
                 sequence 
               
               
                   
               
               
                 gRNA1 
                 TAGTCTTAGA 
                 UAGUCUUAGAGU 
               
               
                 promoter 
                 GTATCCAGTG 
                 AUCCAGUG 
               
               
                 HBG1/2 
                 (SEQ ID NO: 43) 
                 (SEQ ID NO: 49) 
               
               
                   
               
               
                 gRNA2 
                 GGCTAGGGATG 
                 GGCUAGGGAU 
               
               
                 promoter 
                 AAGAATAAA 
                 GAAGAAUAAA 
               
               
                 HBG1/2 
                 (SEQ ID NO: 44) 
                 (SEQ ID NO: 50) 
               
               
                   
               
               
                 gRNAI HS2 
                 AATATGTCACA 
                 AAUAUGUCAC 
               
               
                 enhancer 
                 TTCTGTCTC 
                 AUUCUGUCUC 
               
               
                 HBG1/2 
                 (SEQ ID NO: 45) 
                 (SEQ ID NO: 51) 
               
               
                   
               
               
                 gRNA2HS2 
                 GGACTATGGG 
                 GGACUAUGGG 
               
               
                 enhancer 
                 AGGTCACTAA 
                 AGGUCACUAA 
               
               
                 HBG1/2 
                 (SEQ ID NO: 46) 
                 (SEQ ID NO: 52) 
               
               
                   
               
               
                 gRNA3HS2 
                 GAAGGTTACAC 
                 GAAGGUUACA 
               
               
                 enhancer 
                 AGAACCAGA 
                 CAGAACCAGA 
               
               
                 HBG1/2 
                 (SEQ ID NO: 47) 
                 (SEQ ID NO: 53) 
               
               
                   
               
               
                 gRNA4 HS2 
                 GCCCTGTAAGC 
                 GCCCUGUAAGC 
               
               
                 enhancer 
                 ATCCTGCTG 
                 AUCCUGCUG 
               
               
                 HBG1/2 
                 (SEQ ID NO: 48) 
                 (SEQ ID NO: 54) 
               
               
                   
               
            
           
         
       
     
     4. Genetic Constructs 
     The fusion protein or DNA Targeting system or a component thereof may be encoded by or comprised within a genetic construct. The DNA Targeting system may comprise one or more genetic constructs. The genetic construct, such as a plasmid or expression vector, may comprise a nucleic acid that encodes the fusion protein or DNA Targeting system and/or at least one of the gRNAs. In certain embodiments, a genetic construct encodes one gRNA molecule, i.e., a first gRNA molecule, and optionally a Cas9 molecule or fusion protein. In some embodiments, a genetic construct encodes two gRNA molecules, i.e., a first gRNA molecule and a second gRNA molecule, and optionally a Cas9 molecule or fusion protein. In some embodiments, a first genetic construct encodes one gRNA molecule, a first gRNA molecule, and optionally a Cas9 molecule or fusion protein, and a second genetic construct encodes one gRNA molecule, i.e., a second gRNA molecule, and optionally a Cas9 molecule or fusion protein. 
     Genetic constructs may include polynucleotides such as vectors and plasmids. The genetic construct may be a linear minichromosome including centromere, telomeres, or plasmids or cosmids. The vector may be an expression vectors or system to produce protein by routine techniques and readily available starting materials including Sambrook et al., Molecular Cloning and Laboratory Manual, Second Ed., Cold Spring Harbor (1989), which is incorporated fully by reference. The construct may be recombinant. The genetic construct may be part of a genome of a recombinant viral vector, including recombinant lentivirus, recombinant adenovirus, and recombinant adenovirus associated virus. The genetic construct may comprise regulatory elements for gene expression of the coding sequences of the nucleic acid. The regulatory elements may be a promoter, an enhancer, an initiation codon, a stop codon, or a polyadenylation signal. 
     The genetic construct may comprise heterologous nucleic acid encoding the fusion protein or DNA Targeting system and may further comprise an initiation codon, which may be upstream of the fusion protein or DNA Targeting system coding sequence, and a stop codon, which may be downstream of the fusion protein or DNA Targeting system coding sequence. The initiation and termination codon may be in frame with the fusion protein or DNA Targeting system coding sequence. The vector may also comprise a promoter that is operably linked to the fusion protein or DNA Targeting system coding sequence. The promoter may be a constitutive promoter, an inducible promoter, a repressible promoter, or a regulatable promoter. The promoter may be a ubiquitous promoter. The promoter may be a tissue-specific promoter. The tissue specific promoter may be a muscle specific promoter. The tissue specific promoter may be a skin specific promoter. The fusion protein or DNA Targeting system may be under the light-inducible or chemically inducible control to enable the dynamic control of gene/genome editing in space and time. The promoter operably linked to the fusion protein or DNA Targeting system coding sequence may be a promoter from simian virus 40 (SV40), a mouse mammary tumor virus (MMTV) promoter, a human immunodeficiency virus (HIV) promoter such as the bovine immunodeficiency virus (BIV) long terminal repeat (LTR) promoter, a Moloney virus promoter, an avian leukosis virus (ALV) promoter, a cytomegalovirus (CMV) promoter such as the CMV immediate early promoter, Epstein Barr virus (EBV) promoter, or a Rous sarcoma virus (RSV) promoter. The promoter may also be a promoter from a human gene such as human ubiquitin C (hUbC), human actin, human myosin, human hemoglobin, human muscle creatine, or human metalothionein. Examples of a tissue specific promoter, such as a muscle or skin specific promoter, natural or synthetic, are described in U.S. Patent Application Publication No. 0520040175727, the contents of which are incorporated herein in its entirety. The promoter may be a CK8 promoter, a Spc512 promoter, a MHCK7 promoter, for example. 
     The genetic construct may also comprise a polyadenylation signal, which may be downstream of the fusion protein or DNA Targeting system. The polyadenylation signal may be a SV40 polyadenylation signal, LTR polyadenylation signal, bovine growth hormone (bGH) polyadenylation signal, human growth hormone (hGH) polyadenylation signal, or human p-globin polyadenylation signal. The SV40 polyadenylation signal may be a polyadenylation signal from a pCEP4 vector (Invitrogen, San Diego, Calif.). 
     Coding sequences in the genetic construct may be optimized for stability and high levels of expression. In some instances, codons are selected to reduce secondary structure formation of the RNA such as that formed due to intramolecular bonding. 
     The genetic construct may also comprise an enhancer upstream of the fusion protein or DNA Targeting system or gRNAs. The enhancer may be necessary for DNA expression. The enhancer may be human actin, human myosin, human hemoglobin, human muscle creatine or a viral enhancer such as one from CMV, HA, RSV, or EBV. Polynucleotide function enhancers are described in U.S. Pat. Nos. 5,593,972, 5,962,428, and WO94/016737, the contents of each are fully incorporated by reference. The genetic construct may also comprise a mammalian origin of replication in order to maintain the vector extrachromosomally and produce multiple copies of the vector in a cell. The genetic construct may also comprise a regulatory sequence, which may be well suited for gene expression in a mammalian or human cell into which the vector is administered. The genetic construct may also comprise a reporter gene, such as green fluorescent protein (“GFP”) and/or a selectable marker, such as hygromycin (“Hygro”). 
     The genetic construct may be useful for transfecting cells with nucleic acid encoding the fusion protein or DNA Targeting system, which the transformed host cell is cultured and maintained under conditions wherein expression of the fusion protein or DNA Targeting system takes place. The genetic construct may be transformed or transduced into a cell. The genetic construct may be formulated into any suitable type of delivery vehicle including, for example, a viral vector, lentiviral expression, mRNA electroporation, and lipid-mediated transfection for delivery into a cell. The genetic construct may be part of the genetic material in attenuated live microorganisms or recombinant microbial vectors which live in cells. The genetic construct may be present in the cell as a functioning extrachromosomal molecule. 
     Further provided herein is a cell transformed or transduced with a system or component thereof as detailed herein. Suitable cell types are detailed herein. In some embodiments, the cell is a stem cell. The stem cell may be a human stem cell. In some embodiments, the cell is an embryonic stem cell. The stem cell may be a human pluripotent stem cell (iPSCs). Further provided are stem cell-derived neurons, such as neurons derived from iPSCs transformed or transduced with a DNA targeting system or component thereof as detailed herein. 
     a. Viral Vectors 
     A genetic construct may be a viral vector. Further provided herein is a viral delivery system. Viral delivery systems may include, for example, lentivirus, retrovirus, adenovirus, mRNA electroporation, or nanoparticles. In some embodiments, the vector is a modified lentiviral vector. In some embodiments, the viral vector is an adeno-associated virus (AAV) vector. The AAV vector is a small virus belonging to the genus Dependovirus of the Parvoviridae family that infects humans and some other primate species. 
     AAV vectors may be used to deliver fusion proteins or DNA Targeting systems using various construct configurations. For example, AAV vectors may deliver Cas9 or fusion protein and gRNA expression cassettes on separate vectors or on the same vector. Alternatively, if the small Cas9 proteins or fusion proteins, derived from species such as  Staphylococcus  aureus or  Neisseria meningitidis , are used then both the Cas9 and up to two gRNA expression cassettes may be combined in a single AAV vector. In some embodiments, the AAV vector has a 4.7 kb packaging limit. 
     In some embodiments, the AAV vector is a modified AAV vector. The modified AAV vector may have tissue-specific tropism. The modified AAV vector may be capable of delivering and expressing the fusion protein or DNA Targeting system in the cell of a mammal. For example, the modified AAV vector may be an AAV-SASTG vector (Piacentino et al. Human Gene Therapy 2012, 23, 635-646). The modified AAV vector may be based on one or more of several capsid types, including AAV1, AAV2, AAV5, AAV6, AAV8, and AAV9. The modified AAV vector may be based on AAV2 pseudotype with alternative tissue-tropic AAV capsids, such as AAV2/1, AAV2/6, AAV2/7, AAV2/8, AAV2/9, AAV2.5, and AAV/SASTG vectors that efficiently transduce target tissue by systemic and local delivery (Seto et al. Current Gene Therapy 2012, 12, 139-151). The modified AAV vector may be AAV2i8G9 (Shen et al. J. Biol. Churn. 2013, 288, 28814-28823). 
     5. Pharmaceutical Compositions 
     Further provided herein are pharmaceutical compositions comprising the above-described fusion proteins or DNA Targeting systems or genetic constructs. In some embodiments, the pharmaceutical composition may comprise about 1 ng to about 10 mg of DNA encoding the fusion protein or DNA Targeting system. The systems or genetic constructs as detailed herein, or at least one component thereof, may be formulated into pharmaceutical compositions in accordance with standard techniques well known to those skilled in the pharmaceutical art. The pharmaceutical compositions can be formulated according to the mode of administration to be used. In cases where pharmaceutical compositions are injectable pharmaceutical compositions, they are sterile, pyrogen free, and particulate free. An isotonic formulation is preferably used. Generally, additives for isotonicity may include sodium chloride, dextrose, mannitol, sorbitol and lactose. In some cases, isotonic solutions such as phosphate buffered saline are preferred. Stabilizers include gelatin and albumin. In some embodiments, a vasoconstriction agent is added to the formulation. 
     The composition may further comprise a pharmaceutically acceptable excipient. The pharmaceutically acceptable excipient may be functional molecules as vehicles, adjuvants, carriers, or diluents. The term “pharmaceutically acceptable carrier,” may be a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Pharmaceutically acceptable carriers include, for example, diluents, lubricants, binders, disintegrants, colorants, flavors, sweeteners, antioxidants, preservatives, glidants, solvents, suspending agents, wetting agents, surfactants, emollients, propellants, humectants, powders, pH adjusting agents, and combinations thereof. The pharmaceutically acceptable excipient may be a transfection facilitating agent, which may include surface active agents, such as immune-stimulating complexes (ISCOMS), Freunds incomplete adjuvant, LPS analog including monophosphoryl lipid A, muramyl peptides, quinone analogs, vesicles such as squalene and squalene, hyaluronic acid, lipids, liposomes, calcium ions, viral proteins, polyanions, polycations, or nanoparticles, or other known transfection facilitating agents. The transfection facilitating agent may be a polyanion, polycation, including poly-L-glutamate (LGS), or lipid. The transfection facilitating agent may be poly-L-glutamate, and more preferably, the poly-L-glutamate may be present in the composition for gene editing in skeletal muscle or cardiac muscle at a concentration less than 6 mg/mL. 
     6. Administration 
     The systems or genetic constructs as detailed herein, or at least one component thereof, may be administered or delivered to a cell. Methods of introducing a nucleic acid into a host cell are known in the art, and any known method can be used to introduce a nucleic acid (e.g., an expression construct) into a cell. Suitable methods include, for example, viral or bacteriophage infection, transfection, conjugation, protoplast fusion, polycation or lipid:nucleic acid conjugates, lipofection, electroporation, nucleofection, immunoliposomes, calcium phosphate precipitation, polyethyleneimine (PEI)-mediated transfection, DEAE-dextran mediated transfection, liposome-mediated transfection, particle gun technology, calcium phosphate precipitation, direct micro injection, nanoparticle-mediated nucleic acid delivery, and the like. In some embodiments, the composition may be delivered by mRNA delivery and ribonucleoprotein (RNP) complex delivery. The system, genetic construct, or composition comprising the same, may be electroporated using BioRad Gene Pulser Xcell or Amaxa Nucleofector IIb devices or other electroporation device. Several different buffers may be used, including BioRad electroporation solution, Sigma phosphate-buffered saline product #D8537 (PBS), Invitrogen OptiMEM I (OM), or Amaxa Nucleofector solution V (N.V.). Transfections may include a transfection reagent, such as Lipofectamine 2000. 
     The systems or genetic constructs as detailed herein, or at least one component thereof, or the pharmaceutical compositions comprising the same, may be administered to a subject. Such compositions can be administered in dosages and by techniques well known to those skilled in the medical arts taking into consideration such factors as the age, sex, weight, and condition of the particular subject, and the route of administration. The presently disclosed systems, or at least one component thereof, genetic constructs, or compositions comprising the same, may be administered to a subject by different routes including orally, parenterally, sublingually, transdermally, rectally, transmucosally, topically, intranasal, intravaginal, via inhalation, via buccal administration, intrapleurally, intravenous, intraarterial, intraperitoneal, subcutaneous, intradermally, epidermally, intramuscular, intranasal, intrathecal, intracranial, and intraarticular or combinations thereof. In certain embodiments, the system, genetic construct, or composition comprising the same, is administered to a subject intramuscularly, intravenously, or a combination thereof. The systems, genetic constructs, or compositions comprising the same may be delivered to a subject by several technologies including DNA injection (also referred to as DNA vaccination) with and without in vivo electroporation, liposome mediated, nanoparticle facilitated, recombinant vectors such as recombinant lentivirus, recombinant adenovirus, and recombinant adenovirus associated virus. The composition may be injected into the brain or other component of the central nervous system. The composition may be injected into the skeletal muscle or cardiac muscle. For example, the composition may be injected into the tibias anterior muscle or tail. For veterinary use, the systems, genetic constructs, or compositions comprising the same may be administered as a suitably acceptable formulation in accordance with normal veterinary practice. The veterinarian may readily determine the dosing regimen and route of administration that is most appropriate for a particular animal. The systems, genetic constructs, or compositions comprising the same may be administered by traditional syringes, needleless injection devices, “microprojectile bombardment gone guns,” or other physical methods such as electroporation (“EP”), “hydrodynamic method”, or ultrasound. Alternatively, transient in vivo delivery of fusion proteins or DNA Targeting systems by non-viral or non-integrating viral gene transfer, or by direct delivery of purified proteins and gRNAs containing cell-penetrating motifs may enable highly specific correction and/or restoration in situ with minimal or no risk of exogenous DNA integration. 
     Upon delivery of the presently disclosed systems or genetic constructs as detailed herein, or at least one component thereof, or the pharmaceutical compositions comprising the same, and thereupon the vector into the cells of the subject, the transfected cells may express the gRNA molecule(s) and the Cas9 molecule or fusion protein. 
     a. Cell Types 
     Any of the delivery methods and/or routes of administration detailed herein can be utilized with a myriad of cell types. Further provided herein is a cell transformed or transduced with a system or component thereof as detailed herein. For example, provided herein is a cell comprising an isolated polynucleotide encoding a DNA targeting system as detailed herein. Suitable cell types are detailed herein. In some embodiments, the cell is an immune cell. Immune cells may include, for example, lymphocytes such as T cells and B cells and natural killer (NK) cells. In some embodiments, the cell is a T cell. T cells may be divided into cytotoxic T cells and helper T cells, which are in turn categorized as TH1 or TH2 helper T cells. Immune cells may further include innate immune cells, adaptive immune cells, tumor-primed T cells, NIST cells, IFN-γ producing killer dendritic cells (IKDC), memory T cells (TCMs), and effector T cells (TEs). The cell may be a stem cell such as a human stem cell. In some embodiments, the cell is an embryonic stem cell or a hematopoietic stem cell. The stem cell may be a human induced pluripotent stem cell (iPSCs). Further provided are stem cell-derived neurons, such as neurons derived from iPSCs transformed or transduced with a DNA targeting system or component thereof as detailed herein. The cell may be a muscle cell. Cells may further include, but are not limited to, immortalized myoblast cells, dermal fibroblasts, primal dermal fibroblasts, bone marrow-derived progenitors, skeletal muscle progenitors, human skeletal myoblasts, CD 133+ cells, mesoangioblasts, cardiomyocytes, hepatocytes, chondrocytes, mesenchymal progenitor cells, hematopoietic stem cells, smooth muscle cells, dendritic cells, and MyoD- or Pax7-transduced cells, or other myogenic progenitor cells. 
     7. Kits 
     Provided herein is a kit, which may be used to enhance or increase expression of a gene. The kit comprises genetic constructs or a composition comprising the same, as described above, and instructions for using said composition. In some embodiments, the kit comprises at least one fusion protein, and instructions for using the fusion protein. 
     Instructions included in kits may be affixed to packaging material or may be included as a package insert. While the instructions are typically written on printed materials they are not limited to such. Any medium capable of storing such instructions and communicating them to an end user is contemplated by this disclosure. Such media include, but are not limited to, electronic storage media (e.g., magnetic discs, tapes, cartridges, chips), optical media (e.g., CD ROM), and the like. As used herein, the term “instructions” may include the address of an Internet site that provides the instructions. 
     8. Methods 
     a. Methods of Activating Expression of a Target Gene 
     Provided herein are methods of activating expression of a target gene in a cell. The methods may include contacting the cell with a fusion protein as detailed herein or a DNA Targeting System as detailed herein or a gRNA as detailed herein. In some embodiments, the target gene is a gamma globin gene. In some embodiments, the gene is gamma globin genes 1 and 2 (HBG1/2). In some embodiments, the DNA Targeting System includes at least one gRNA corresponding to at least one of SEQ ID NOs: 43-54 as detailed herein. The DNA Targeting system may target, for example, the promoter of HBG1/2 and/or the HS2 enhancer region of HBG1/2. In some embodiments, one or two gRNAs are used to target the promoter of HBG1/2 to activate its expression. For example, the DNA Targeting System may include one or two gRNAs that are encoded by or target a sequence of SEQ ID NO: 43 and/or 44, to target the promoter of HBG1/2 to activate its expression. The DNA Targeting System may include one or two gRNAs comprising a polynucleotide sequence selected from SEQ ID NOs: 49 and 50, to target the promoter of HBG1/2 to activate its expression. In some embodiments, four gRNAs are used to target the HS2 enhancer region of HBG1/2 to activate its expression. For example, the DNA Targeting System may include one, two, three, or four gRNAs that are encoded by or target a sequence selected from SEQ ID NOs: 45-48, to target the HS2 enhancer region of HBG1/2 to activate its expression. The DNA Targeting System may include one, two, three, or four gRNAs comprising a polynucleotide sequence selected from SEQ ID NOs: 51-54, to target the HS2 enhancer region of HBG1/2 to activate its expression. In some embodiments, methods disclosed herein increase mRNA expression of the target gene in a cell compared to a control. The control may be the mRNA expression of the target gene in a cell in which the fusion protein is not present. In some embodiments, methods disclosed herein increase the level of protein expressed from the target gene in a cell compared to a control. The control may be the level of protein expression from the target gene in a cell in which the fusion protein is not present. 
     9. Examples 
     The foregoing may be better understood by reference to the following examples, which are presented for purposes of illustration and are not intended to limit the scope of the invention. The present disclosure has multiple aspects and embodiments, illustrated by the appended non-limiting examples. 
     Example 1 
     Combining Chromatin Remodelers with Activator Domains Enhances Activation of Target Gene Expression 
     Targeted activation of endogenous genes with synthetic transcription factors or epigenome editors, made from DNA-targeting systems such as zinc finger proteins, TALEs, and CRISPR-Cas systems, are broadly useful for gene therapy, regenerative medicine, and programming stem cell differentiation. However, a common limitation is that the potency of gene activation is insufficient to generate the desired phenotype or biological effect. Here, it is demonstrated that combining modulators of chromatin structure (for example, SS18 or the SWI/SNF (BAF) chromatin remodeling complex) with activator domains (for example, VP64, VPH, VPR, and/or p300) can lead to more potent gene activation in human cells. 
     In an effort to generate more potent transcriptional activators relative to state-of-the-art by rational design of programmable gene modulators, the potency of dCas9-VPH to activate the HBG1/2 gene relative to p300core (p300c) and VPR fusions was compared ( FIG.  1 A ). VPH, a fusion of VP64, mouse p65 activation domain (AD), and HSF1 (AD), has been used in few instances but is less commonly used than VPR (VP64, human p65 activation domain (AD), and Rta) ( FIG.  18   ). All three effectors lead to deposition of histone H3K27 acetylation, a mark of gene activation, as well as recruitment of transcription factors. dCas9-VPH was more potent than p300c or VPR in activating HBG1/2 when targeted to either its promoter or enhancer ( FIG.  1 C ). 
     The SWI/SNF (BAF) chromatin remodeling complex has been shown to antagonize PRC1/2 complexes that deposit and bind to the repressive H3K27 trimethylation and H2A ubiquitylation histone marks, leading to a more facultative chromatin state. To design a more potent dCas9 activator, the potency of dCas9-VPH or dCas9-p300c activating HBG1/2 when used in combination with a dCas9-SS18 fusion was examined, since the SS18 subunit of the BAF complex is sufficient to recruit the full BAF complex to chromatin. Using dCas9-SS18 in combination with dCas9-VPH showed greater activation of HBG1/2 compared with that achieved by its combination with dCas9-p300c or any of the dCas9-fusions alone ( FIG.  2   ). Fusion of several transcriptional regulators to dCas9 in tandem can lead to a synergistic increase in activity. To generate a dCas9 bipartite activator consisting of VPH and SS18, it was examined whether VPH was a more potent activator when fused to dCas9 at its N-terminus or C-terminus. dCas9 fused to p300c or to two VP64 domains (one on each termini) were also included as controls. VPH showed the strongest activation of HBG1/2 when fused at the N-terminus of dCas9 and was the most potent of all the activators tested ( FIG.  3   ). For the design of the bipartite dCas9 activator, VPH was fused on the N-terminus of dCas9 and SS18 on its C-terminus. When tested for HBG1/2 activation in a side-by-side comparison of the best dCas9-fusion activators, the VPH-dCas9-SS18 bipartite activator outperformed both p300c and VPH as a single fusion to dCas9 ( FIG.  4   ). 
     Collectively, these results support a model in which remodelers of chromatin structure cooperate with co-recruited transcriptional activation domains to more robustly activate target gene expression. 
     The foregoing description of the specific aspects will so fully reveal the general nature of the invention that others can, by applying knowledge within the skill of the art, readily modify and/or adapt for various applications such specific aspects, without undue experimentation, without departing from the general concept of the present disclosure. Therefore, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed aspects, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance. 
     The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary aspects but should be defined only in accordance with the following claims and their equivalents. 
     All publications, patents, patent applications, and/or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, and/or other document were individually indicated to be incorporated by reference for all purposes. 
     For reasons of completeness, various aspects of the invention are set out in the following numbered clauses: 
     Clause 1. A fusion protein comprising at least two heterologous polypeptide domains, wherein the first polypeptide domain comprises a DNA binding protein and the second polypeptide domain comprises a modulator of chromatin structure. 
     Clause 2. The fusion protein of clause 1, wherein the fusion protein further comprises a third polypeptide domain. 
     Clause 3. The fusion protein of any one of the preceding clauses, wherein the first polypeptide domain comprises a CRISPR-associated (Gas) protein, a TALE, or a zinc finger protein. 
     Clause 4. The fusion protein of clause 3, wherein the Cas protein comprises at least one amino acid mutation that eliminates nuclease activity of the Cas protein. 
     Clause 5. The fusion protein of clause 3 or 4, wherein the Cas protein comprises a Cas9 protein. 
     Clause 6. The fusion protein of clause 5, wherein the Cas9 protein is nuclease-deficient dCas9 and comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 20 or 21 or is encoded by a polynucleotide comprising a sequence having at least 75% identity to SEQ ID NO: 22 or 23. 
     Clause 7. The fusion protein of any one of the preceding clauses, wherein the modulator of chromatin structure comprises a nucleosome rearranging protein. 
     Clause 8. The fusion protein of any one of the preceding clauses, wherein the modulator of chromatin structure comprises the SS18 subunit of the BAF chromatin remodeling complex or a fragment thereof or a variant thereof. 
     Clause 9. The fusion protein of clause 8, wherein the SS18 subunit comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 37. 
     Clause 10. The fusion protein of any one of clauses 2-9, wherein the third polypeptide domain comprises a transcriptional activator domain. 
     Clause 11. The fusion protein of clause 10, wherein the transcriptional activator domain comprises VP64, VPH, VPR, p65, TET1, or p300, or a combination thereof or a fragment thereof or a variant thereof. 
     Clause 12. The fusion protein of clause 11, wherein the VP64 comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 91. 
     Clause 13. The fusion protein of clause 11, wherein the TET1 comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 93. 
     Clause 14. The fusion protein of clause 11, wherein the VPH comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 39. 
     Clause 15. The fusion protein of clause 11, wherein the VPR comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 41. 
     Clause 16. The fusion protein of clause 11, wherein the p300 comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 33 or 34. 
     Clause 17. The fusion protein of any one of clauses 1-16, wherein the fusion protein comprises one or more second polypeptide domain(s). 
     Clause 18. The fusion protein of clause 17, wherein the one or more second polypeptide domain(s) is fused to the C-terminus or the N-terminus of the first polypeptide domain, or a combination thereof. 
     Clause 19. The fusion protein of clause 18, wherein the N-terminus of the second polypeptide is operably linked to the C-terminus of the first polypeptide domain, or wherein the C-terminus of the second polypeptide is operably linked to the N-terminus of the first polypeptide domain. 
     Clause 20. The fusion protein of any one of clauses 2-19, wherein the fusion protein comprises one or more third polypeptide domain(s). 
     Clause 21. The fusion protein of clause 20, wherein the one or more third polypeptide domain is fused to the C-terminus or the N-terminus of the first polypeptide domain, or a combination thereof. 
     Clause 22. The fusion protein of clause 21, wherein the N-terminus of the third polypeptide is operably linked to the C-terminus of the first polypeptide domain, or wherein the C-terminus of the third polypeptide is operably linked to the N-terminus of the first polypeptide domain. 
     Clause 23. The fusion protein of any one of clauses 2-22, wherein the first polypeptide domain comprises dCas9, wherein the second polypeptide domain comprises SS18, and wherein the third polypeptide domain comprises VPH. 
     Clause 24. The fusion protein of clause 23, wherein the fusion protein comprises VPH-dCas9-SS18 or SS18-dCas9-VPH or variants thereof. 
     Clause 25. The fusion protein of clause 24, wherein the fusion protein comprises a polypeptide having at least 75% sequence identity to SEQ ID NO: 64 or 66. 
     Clause 26. The fusion protein of any one of clauses 2-22, wherein the first polypeptide domain comprises dCas9, wherein the second polypeptide domain comprises 8518, and wherein the third polypeptide domain comprises VPR. 
     Clause 27. The fusion protein of clause 26, wherein the fusion protein comprises VPR-dCas9-SS18 or 8518-dCas9-VPR or variants thereof. 
     Clause 28. The fusion protein of any one of clauses 2-22, wherein the first polypeptide domain comprises dCas9, wherein the second polypeptide domain comprises SS18, and wherein the third polypeptide domain comprises p300. 
     Clause 29. The fusion protein of clause 28, wherein the fusion protein comprises p300-dCas9-SS18 or SS18-dCas9-p300 or variants thereof. 
     Clause 30. The fusion protein of any one of clauses 2-22, wherein the first polypeptide domain comprises dCas9, wherein the second polypeptide domain comprises SS18, and wherein the third polypeptide domain comprises VP64. 
     Clause 31. The fusion protein of clause 30, wherein the fusion protein comprises VP64-dCas9-SS18 or SS18-dCas9-VP64 or variants thereof. 
     Clause 32. The fusion protein of any one of the preceding clauses, wherein the fusion protein activates transcription of a target gene. 
     Clause 33. The fusion protein of any one of the preceding clauses, wherein the fusion protein increases the level of mRNA expression of a target gene in a cell containing the fusion protein relative to a control. 
     Clause 34. The fusion protein of clause 33, wherein the level of mRNA expression of the target gene is increased at least 5-fold, at least 50-fold, at least 100-fold, at least 1,000-fold, at least 10,000-fold, or at least 20,000-fold relative to a control. 
     Clause 35. The fusion protein of clause 33 or 34, wherein the level of mRNA expression of the target gene is increased by 5-fold to 10,000-fold, 5-fold to 30,000-fold, 5-fold to 50,000-fold, 5-fold to 100,000-fold, 10,000-fold to 30,000-fold, 20,000-fold to 30,000-fold, 15,000-fold to 25,000-fold, 1,000-fold to 50,000-fold, or 1,000-fold to 100,000-fold relative to a control. 
     Clause 36. The fusion protein of any one of clauses 33-35, wherein the control is the level of mRNA expression of the target gene in a cell not containing the fusion protein. 
     Clause 37. The fusion protein of any one of clauses 32-36, wherein the target gene is gamma globin genes 1 and 2 (HBG1/2). 
     Clause 38. A DNA Targeting System comprising: (a) the fusion protein of any one of clauses 1-37, wherein the first polypeptide domain comprises a zinc finger protein or a TALE; or (b) a gRNA and the fusion protein of any one of clauses 1-37, wherein the first polypeptide domain comprises a Cas protein, and wherein the gRNA targets a target gene. 
     Clause 39. The DNA Targeting System of clause 38, wherein gRNA targets a regulatory region of the target gene. 
     Clause 40. The DNA Targeting System of clause 39, wherein the regulatory region is a promoter sequence of the target gene. 
     Clause 41. A DNA Targeting System comprising a gRNA that recruits a modulator of chromatin structure to a target sequence. 
     Clause 42. The DNA Targeting System of clause 41, wherein the modulator of chromatin structure comprises the SS18 subunit of the BAF chromatin remodeling complex. 
     Clause 43. The DNA Targeting System of any one of clauses 38-42, wherein the gRNA is encoded by or binds to a target sequence selected from SEQ ID NOs: 43-48, a complement thereof, a truncation thereof, or a variant thereof, or wherein the gRNA is encoded by or binds to a target sequence having at least 70% sequence identity to a sequence selected from SEQ ID NOs: 43-48, a complement thereof, a truncation thereof, or a variant thereof. 
     Clause 44. The DNA Targeting System of any one of clauses 38-43, wherein the gRNA comprises a polynucleotide sequence selected from SEQ ID NOs: 49-54, a complement thereof, a truncation thereof, or a variant thereof, or wherein the gRNA comprises a polynucleotide having at least 70% sequence identity to a sequence selected from SEQ ID NOs: 49-54, a complement thereof, a truncation thereof, or a variant thereof. 
     Clause 45. A method of increasing expression of a target gene in a cell, the method comprising contacting the cell with the fusion protein of any one of clauses 1-37 or the DNA Targeting system of any one of clauses 38-44. 
     Clause 46. The method of clause 45, wherein the target gene is gamma globin genes 1 and 2 (HBG1/2). 
     Clause 47. A gRNA encoded by or binding to a target sequence selected from SEQ ID NOs: 43-48, a complement thereof, a truncation thereof, or a variant thereof, or comprising a polynucleotide sequence selected from SEQ ID NOs: 49-54, a complement thereof, a truncation thereof, or a variant thereof. 
     
       
         
           
               
               
            
               
                 SEQUENCES 
                   
               
               
                 SEQ ID NO: 1 
                   
               
               
                 NRG 
                   
               
               
                 (R = A or G; N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 2 
                   
               
               
                 NGG 
                   
               
               
                 (N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 3 
                   
               
               
                 NAG 
                   
               
               
                 (N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 4 
                   
               
               
                 NGGNG 
                   
               
               
                 (N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 5 
                   
               
               
                 NNAGAAW 
                   
               
               
                 (W = A or T; N can be any nucleotide residue, e.g,, any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 6 
                   
               
               
                 NAAR 
                   
               
               
                 (R = A or G; N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 7 
                   
               
               
                 NNGRR 
                   
               
               
                 (R = A or G; N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 8 
                   
               
               
                 NNGRRN 
                   
               
               
                 (R = A or G; N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 9 
                   
               
               
                 NNGRRT 
                   
               
               
                 (R = A or G; N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 10 
                   
               
               
                 NNGRRV 
                   
               
               
                 (R = A or G; N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 11 
                   
               
               
                 NNNNGATT 
                   
               
               
                 (N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 12 
                   
               
               
                 NNNNGNNN 
                   
               
               
                 (N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 13 
                   
               
               
                 NGA 
                   
               
               
                 (N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 14 
                   
               
               
                 NNNRRT 
                   
               
               
                 (R = A or G; N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 15 
                   
               
               
                 ATTCCT 
                   
               
               
                   
               
               
                 SEQ ID NO: 16 
                   
               
               
                 NGAN 
                   
               
               
                 (N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                 SEQ ID NO: 17 
                   
               
               
                 NGNG 
                   
               
               
                 (N can be any nucleotide residue, e.g., any of A, G, C, or T) 
               
               
                   
               
               
                   Streptococcus pyogenes  Cas9 
               
               
                 SEQ ID NO: 18 
                   
               
               
                 MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                   
               
               
                   
               
               
                 RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIY 
               
               
                   
               
               
                 HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
               
               
                   
               
               
                 GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
               
               
                   
               
               
                 DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
               
               
                   
               
               
                 QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
               
               
                   
               
               
                 SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
               
               
                   
               
               
                 NFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQ 
               
               
                   
               
               
                 KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
               
               
                   
               
               
                 EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
               
               
                   
               
               
                 DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
               
               
                   
               
               
                 KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
               
               
                   
               
               
                 QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
               
               
                   
               
               
                 QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
               
               
                   
               
               
                 VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
               
               
                   
               
               
                 MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
               
               
                   
               
               
                 MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKK 
               
               
                   
               
               
                 LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
               
               
                   
               
               
                 ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLS 
               
               
                   
               
               
                 AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
               
               
                   
               
               
                 DLSQLGGD 
               
               
                   
               
               
                   Staphylococcus aureus  Cas9 molecule 
               
               
                 SEQ ID NO: 19 
                   
               
               
                 MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQRVK 
                   
               
               
                   
               
               
                 KLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDTGNELSTKE 
               
               
                   
               
               
                 QISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDL 
               
               
                   
               
               
                 LETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDEN 
               
               
                   
               
               
                 EKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE 
               
               
                   
               
               
                 IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELW 
               
               
                   
               
               
                 HTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIII 
               
               
                   
               
               
                 ELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLE 
               
               
                   
               
               
                 DLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLA 
               
               
                   
               
               
                 KGKGRISKTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGF 
               
               
                   
               
               
                 TSFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQ 
               
               
                   
               
               
                 EYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKL 
               
               
                   
               
               
                 KKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYG 
               
               
                   
               
               
                 NKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEAKK 
               
               
                   
               
               
                 LKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPPRIIKTI 
               
               
                   
               
               
                 ASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKG 
               
               
                   
               
               
                   Streptococcus pyogenes  Cas9 
               
               
                 (with D10A) 
               
               
                 SEQ ID NO: 20 
                   
               
               
                 MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                   
               
               
                   
               
               
                 RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIY 
               
               
                   
               
               
                 HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
               
               
                   
               
               
                 GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
               
               
                   
               
               
                 DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
               
               
                   
               
               
                 QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
               
               
                   
               
               
                 SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
               
               
                   
               
               
                 NFEEVVDKGASAQSFIERMTNFDKKLPNEKVLPKHSLLYEYFTVYMELTKVKYVTEGMRKPAFLSGEQ 
               
               
                   
               
               
                 KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
               
               
                   
               
               
                 EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
               
               
                   
               
               
                 DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
               
               
                   
               
               
                 KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
               
               
                   
               
               
                 QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
               
               
                   
               
               
                 QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
               
               
                   
               
               
                 VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
               
               
                   
               
               
                 MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
               
               
                   
               
               
                 MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKK 
               
               
                   
               
               
                 LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
               
               
                   
               
               
                 ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLS 
               
               
                   
               
               
                 AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
               
               
                   
               
               
                 DLSQLGGD 
               
               
                   
               
               
                   Streptococcus pyogenes  Cas9 
               
               
                 (with D10A, H849A) 
               
               
                 SEQ ID NO: 21 
                   
               
               
                 MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                   
               
               
                   
               
               
                 RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIY 
               
               
                   
               
               
                 HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
               
               
                   
               
               
                 GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
               
               
                   
               
               
                 DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
               
               
                   
               
               
                 QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
               
               
                   
               
               
                 SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
               
               
                   
               
               
                 NFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQ 
               
               
                   
               
               
                 KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
               
               
                   
               
               
                 EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
               
               
                   
               
               
                 DFLKSDGFANRNEMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
               
               
                   
               
               
                 KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
               
               
                   
               
               
                 QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
               
               
                   
               
               
                 QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
               
               
                   
               
               
                 VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
               
               
                   
               
               
                 MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIBTNGETGEIVWDKGRDFATVRKVLS 
               
               
                   
               
               
                 MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKK 
               
               
                   
               
               
                 LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
               
               
                   
               
               
                 ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLS 
               
               
                   
               
               
                 AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
               
               
                   
               
               
                 DLSQLGGD 
               
               
                   
               
               
                 Polynucleotide sequence of D10A mutant of  S. aureus  Cas9 
               
               
                 SEQ ID NO: 22 
                   
               
               
                 atgaaaagga actacattct ggggctggcc atcgggatta caagcgtggg gtatgggatt 
                   
               
               
                   
               
               
                 attgactatg aaacaaggga cgtgatcgac gcaggcgtca gactgttcaa ggaggccaac 
               
               
                   
               
               
                 gtggaaaaca atgagggacg gagaagcaag aggggagcca ggcgcctgaa acgacggaga 
               
               
                   
               
               
                 aggcacagaa tccagagggt gaagaaactg ctgttcgatt acaacctgct gaccgaccat 
               
               
                   
               
               
                 tctgagctga gtggaattaa tccttatgaa gccagggtga aaggcctgag tcagaagctg 
               
               
                   
               
               
                 tcagaggaag agttttccgc agctctgctg cacctggcta agcgccgagg agtgcataac 
               
               
                   
               
               
                 gtcaatgagg tggaagagga caccggcaac gagctgtcta caaaggaaca gatctcacgc 
               
               
                   
               
               
                 aatagcaaag ctctggaaga gaagtatgtc gcagagctgc agctggaacg gctgaagaaa 
               
               
                   
               
               
                 gatggcgagg tgagagggtc aactaatagg ttcaagacaa gcgactacgt caaagaagcc 
               
               
                   
               
               
                 aagcagctgc tgaaagtgca gaaggcttac caccagctgg atcagagctt catcgatact 
               
               
                   
               
               
                 tatatcgacc tgctggagac tcggagaacc tactatgagg gaccaggaga agggagcccc 
               
               
                   
               
               
                 ttcggatgga aagacatcaa ggaatggtac gagatgctga tgggacattg cacctatttt 
               
               
                   
               
               
                 ccagaagagc tgagaagcgt caagtacgct tataacgcag atctgtacaa cgccctgaat 
               
               
                   
               
               
                 gacctgaaca acctggtcat caccagggat gaaaacgaga aactggaata ctatgagaag 
               
               
                   
               
               
                 ttccagatca tcgaaaacgt gtttaagcag aagaaaaagc ctacactgaa acagattgct 
               
               
                   
               
               
                 aaggagatcc tggtcaacga agaggacatc aagggctacc gggtgacaag cactggaaaa 
               
               
                   
               
               
                 ccagagttca ccaatctgaa agtgtatcac gatattaagg acatcacagc acggaaagaa 
               
               
                   
               
               
                 atcattgaga acgccgaact gctggatcag attgctaaga tcctgactat ctaccagagc 
               
               
                   
               
               
                 tccgaggaca tccaggaaga gctgactaac ctgaacagcg agctgaccca ggaagagatc 
               
               
                   
               
               
                 gaacagatta gtaatctgaa ggggtacacc ggaacacaca acctgtccct gaaagctatc 
               
               
                   
               
               
                 aatctgattc tggatgagct gtggcataca aacgacaatc agattgcaat ctttaaccgg 
               
               
                   
               
               
                 ctgaagctgg tcccaaaaaa ggtggacctg agtcagcaga aagagatccc aaccacactg 
               
               
                   
               
               
                 gtggacgatt tcattctgtc acccgtggtc aagcggagct tcatccagag catcaaagtg 
               
               
                   
               
               
                 atcaacgcca tcatcaagaa gtacggcctg cccaatgata tcattatcga gctggctagg 
               
               
                   
               
               
                 gagaagaaca gcaaggacgc acagaagatg atcaatgaga tgcagaaacg aaaccggcag 
               
               
                   
               
               
                 accaatgaac gcattgaaga gattatccga actaccggga aagagaacgc aaagtacctg 
               
               
                   
               
               
                 attgaaaaaa tcaagctgca cgatatgcag gagggaaagt gtctgtattc tctggaggcc 
               
               
                   
               
               
                 atccccctgg aggacctgct gaacaatcca ttcaactacg aggtcgatca tattatcccc 
               
               
                   
               
               
                 agaagcgtgt ccttcgacaa ttcctttaac aacaaggtgc tggtcaagca ggaagagaac 
               
               
                   
               
               
                 tctaaaaagg gcaataggac tcctttccag tacctgtcta gttcagattc caagatctct 
               
               
                   
               
               
                 tacgaaacct ttaaaaagca cattctgaat ctggccaaag gaaagggccg catcagcaag 
               
               
                   
               
               
                 accaaaaagg agtacctgct ggaagagcgg gacatcaaca gattctccgt ccagaaggat 
               
               
                   
               
               
                 tttattaacc ggaatctggt ggacacaaga tacgctactc gcggcctgat gaatctgctg 
               
               
                   
               
               
                 cgatcctatt tccgggtgaa caatctggat gtgaaagtca agtccatcaa cggcgggttc 
               
               
                   
               
               
                 acatcttttc tgaggcgcaa atggaagttt aaaaaggagc gcaacaaagg gtacaagcac 
               
               
                   
               
               
                 catgccgaag atgctctgat tatcgcaaat gccgacttca tctttaagga gtggaaaaag 
               
               
                   
               
               
                 ctggacaaag ccaagaaagt gatggagaac cagatgttcg aagagaagca ggccgaatct 
               
               
                   
               
               
                 atgcccgaaa tcgagacaga acaggagtac aaggagattt tcatcactcc tcaccagatc 
               
               
                   
               
               
                 aagcatatca aggatttcaa ggactacaag tactctcacc gggtggataa aaagcccaac 
               
               
                   
               
               
                 agagagctga tcaatgacac cctgtatagt acaagaaaag acgataaggg gaataccctg 
               
               
                   
               
               
                 attgtgaaca atctgaacgg actgtacgac aaagataatg acaagctgaa aaagctgatc 
               
               
                   
               
               
                 aacaaaagtc ccgagaagct gctgatgtac caccatgatc ctcagacata tcagaaactg 
               
               
                   
               
               
                 aagctgatta tggagcagta cggcgacgag aagaacccac tgtataagta ctatgaagag 
               
               
                   
               
               
                 actgggaact acctgaccaa gtatagcaaa aaggataatg gccccgtgat caagaagatc 
               
               
                   
               
               
                 aagtactatg ggaacaagct gaatgcccat ctggacatca cagacgatta ccctaacagt 
               
               
                   
               
               
                 cgcaacaagg tggtcaagct gtcactgaag ccatacagat tcgatgtcta tctggacaac 
               
               
                   
               
               
                 ggcgtgtata aatttgtgac tgtcaagaat ctggatgtca tcaaaaagga gaactactat 
               
               
                   
               
               
                 gaagtgaata gcaagtgcta cgaagaggct aaaaagctga aaaagattag caaccaggca 
               
               
                   
               
               
                 gagttcatcg cctcctttta caacaacgac ctgattaaga tcaatggcga actgtatagg 
               
               
                   
               
               
                 gtcatcgggg tgaacaatga tctgctgaac cgcattgaag tgaatatgat tgacatcact 
               
               
                   
               
               
                 taccgagagt atctggaaaa catgaatgat aagcgccccc ctcgaattat caaaacaatt 
               
               
                   
               
               
                 gcctctaaga ctcagagtat caaaaagtac tcaaccgaca ttctgggaaa cctgtatgag 
               
               
                   
               
               
                 gtgaagagca aaaagcaccc tcagattatc aaaaagggc 
               
               
                   
               
               
                 Polynucleotide sequence of N580A mutant of  S. aureus  Cas9 
               
               
                 SEQ ID NO: 23 
                   
               
               
                 atgaaaagga actacattct ggggctggac atcgggatta caagcgtggg gtatgggatt 
                   
               
               
                   
               
               
                 attgactatg aaacaaggga cgtgatcgac gcaggcgtca gactgttcaa ggaggccaac 
               
               
                   
               
               
                 gtggaaaaca atgagggacg gagaagcaag aggggagcca ggcgcctgaa acgacggaga 
               
               
                   
               
               
                 aggcacagaa tccagagggt gaagaaactg ctgttcgatt acaacctgct gaccgaccat 
               
               
                   
               
               
                 tctgagctga gtggaattaa tccttatgaa gccagggtga aaggcctgag tcagaagctg 
               
               
                   
               
               
                 tcagaggaag agttttccgc agctctgctg cacctggcta agcgccgagg agtgcataac 
               
               
                   
               
               
                 gtcaatgagg tggaagagga caccggcaac gagctgtcta caaaggaaca gatctcacgc 
               
               
                   
               
               
                 aatagcaaag ctetggaaga gaagtatgtc gcagagctgc agctggaacg gctgaagaaa 
               
               
                   
               
               
                 gatggcgagg tgagagggtc aattaatagg ttcaagacaa gcgactacgt caaagaagcc 
               
               
                   
               
               
                 aagcagctgc tgaaagtgca gaaggcttac caccagctgg atcagagctt catcgatact 
               
               
                   
               
               
                 tatatcgacc tgctggagac tcggagaacc tactatgagg gaccaggaga agggagcccc 
               
               
                   
               
               
                 ttcggatgga aagacatcaa ggaatggtac gagatgctga tgggacattg cacctatttt 
               
               
                   
               
               
                 ccagaagagc tgagaagcgt caagtacgct tataacgcag atctgtacaa cgccctgaat 
               
               
                   
               
               
                 gacctgaaca acctggtcat caccagggat gaaaacgaga aactggaata ctatgagaag 
               
               
                   
               
               
                 ttccagatca tcgaaaacgt gtttaageag aagaaaaagc ctacactgaa acagattgct 
               
               
                   
               
               
                 aaggagatcc tggtcaacga agaggacatc aagggctacc gggtgacaag cactggaaaa 
               
               
                   
               
               
                 ccagagttca ccaatctgaa agtgtatcac gatattaagg acatcacagc acggaaagaa 
               
               
                   
               
               
                 atcattgaga acgccgaact gctggatcag attgctaaga tcctgactat ctaccagagc 
               
               
                   
               
               
                 tccgaggaca tccaggaaga gctgactaac ctgaacagcg agctgaccca ggaagagatc 
               
               
                   
               
               
                 gaacagatta gtaatctgaa ggggtacacc ggaacacaca acctgtccct gaaagctatc 
               
               
                   
               
               
                 aatctgattc tggatgagct gtggcataca aacgacaatc agattgcaat ctttaaccgg 
               
               
                   
               
               
                 ctgaagctgg tcccaaaaaa ggtggacctg agtcagcaga aagagatccc aaccacactg 
               
               
                   
               
               
                 gtggacgatt tcattctgtc acccgtggtc aagcggagct tcatccagag catcaaagtg 
               
               
                   
               
               
                 atcaacgcca tcatcaagaa gtacggcctg cccaatcata tcattatcga gctggctagg 
               
               
                   
               
               
                 gagaagaaca gcaaggacgc acagaagatg atcaatgaga tgcagaaacg aaaccggcag 
               
               
                   
               
               
                 accaatgaac gcattgaaga gattatccga actaccggga aagagaacgc aaagtacctg 
               
               
                   
               
               
                 attgaaaaaa tcaagctgca cgatatgcag gagggaaagt gtctgtattc tctggaggcc 
               
               
                   
               
               
                 atccccctgg aggacctgct gaacaatcca ttcaactacg aggtcgatca tattatcccc 
               
               
                   
               
               
                 agaagcgtgt ccttcgacaa ttcctttaac aacaaggtgc tggtcaagca ggaagaggcc 
               
               
                   
               
               
                 tctaaaaagg gcaataggac tcctttccag tacctgtcta gttcagattc caagatctct 
               
               
                   
               
               
                 tacgaaacct ttaaaaagca cattctgaat ctggccaaag gaaagggccg catcagcaag 
               
               
                   
               
               
                 accaaaaagg agtacctgct ggaagagcgg gacatcaaca gattctccgt ccagaaggat 
               
               
                   
               
               
                 tttattaacc ggaatctggt ggacacaaga tacgctactc gcggcctgat gaatctgctg 
               
               
                   
               
               
                 cgatcctatt tccgggtgaa caatctggat gtgaaagtca agtccatcaa cggcgggttc 
               
               
                   
               
               
                 acatcttttc tgaggcgcaa atggaagttt aaaaaggagc gcaacaaagg gtacaagcac 
               
               
                   
               
               
                 catgccgaag atgctctgat tatcgcaaat gccgacttca tctttaagga gtggaaaaag 
               
               
                   
               
               
                 ctggacaaag ccaagaaagt gatggagaac cagatgttcg aagagaagca ggccgaatct 
               
               
                   
               
               
                 atgcccgaaa tcgagacaga acaggagtac aaggagattt tcatcactcc tcaccagatc 
               
               
                   
               
               
                 aagcatatca aggatttcaa ggactacaag tactctcacc gggtggataa aaagcccaac 
               
               
                   
               
               
                 agagagctga tcaatgacac cctgtatagt acaagaaaag acgataaggg gaataccctg 
               
               
                   
               
               
                 attgtgaaca atctgaacgg actgtacgac aaagataatg acaagctgaa aaagctgatc 
               
               
                   
               
               
                 aacaaaagtc ccgagaagct gctgatgtac caccatgatc ctcagacata tcagaaactg 
               
               
                   
               
               
                 aagctgatta tggagcagta cggcgacgag aagaacccac tgtataagta ctatgaagag 
               
               
                   
               
               
                 actgggaact acctgaccaa gtatagcaaa aaggataatg gccccgtgat caagaagatc 
               
               
                   
               
               
                 aagtactatg ggaacaagct gaatgcccat ctggacatca cagacgatta ccctaacagt 
               
               
                   
               
               
                 cgcaacaagg tggtcaagct gtcactgaag ccatacagat tcgatgtcta tctggacaac 
               
               
                   
               
               
                 ggcgtgtata aatttgtgac tgtcaagaat ctggatgtca tcaaaaagga gaactactat 
               
               
                   
               
               
                 gaagtgaata gcaagtgcta cgaagaggct aaaaagctga aaaagattag caaccaggca 
               
               
                   
               
               
                 gagttcatcg cctcctttta caacaacgac ctgattaaga tcaatggcga actgtatagg 
               
               
                   
               
               
                 gtcatcgggg tgaacaatga tctgctgaac cgcattgaag tgaatatgat tgacatcact 
               
               
                   
               
               
                 taccgagagt atctggaaaa catgaatgat aagcgccccc ctcgaattat caaaacaatt 
               
               
                   
               
               
                 gcctctaaga ctcagagtat caaaaagtac tcaaccgaca ttctgggaaa cctgtatgag 
               
               
                   
               
               
                 gtgaagagca aaaagcaccc tcagattatc aaaaagggc 
               
               
                   
               
               
                 codon optimized polynucleotide encoding S. pyogenes Cas9 
               
               
                 SEQ ID NO: 24 
                   
               
               
                 atggataaaa agtacagcat cgggctggac atcggtacaa actcagtggg gtgggccgtg 
                   
               
               
                   
               
               
                 attacggacg agtacaaggt accctccaaa aaatttaaag tgctgggtaa cacggacaga 
               
               
                   
               
               
                 cactctataa agaaaaatct tattggagcc ttgctgttcg actcaggcga gacagccgaa 
               
               
                   
               
               
                 gccacaaggt tgaagcggac cgccaggagg cggtatacca ggagaaagaa ccgcatatgc 
               
               
                   
               
               
                 tacctgcaag aaatcttcag taacgagatg gcaaaggttg acgatagctt tttccatcgc 
               
               
                   
               
               
                 ctggaagaat cctttcttgt tgaggaagac aagaagcacg aacggcaccc catctttggc 
               
               
                   
               
               
                 aatattgtcg acgaagtggc atatcacgaa aagtacccga ctatctacca cctcaggaag 
               
               
                   
               
               
                 aagctggtgg actctaccga taaggcggac ctcagactta tttatttggc actcgcccac 
               
               
                   
               
               
                 atgattaaat ttagaggaca tttcttgatc gagggcgacc tgaacccgga caacagtgac 
               
               
                   
               
               
                 gtcgataagc tgttcatcca acttgtgcag acctacaatc aactgttcga agaaaaccct 
               
               
                   
               
               
                 ataaatgctt caggagtcga cgctaaagca atcctgtccg cgcgcctctc aaaatctaga 
               
               
                   
               
               
                 agacttgaga atctgattgc tcagttgccc ggggaaaaga aaaatggatt gtttggcaac 
               
               
                   
               
               
                 ctgatcgccc tcagtctcgg actgacccca aatttcaaaa gtaacttcga cctggccgaa 
               
               
                   
               
               
                 gacgctaagc tccagctgtc caaggacaca tacgatgacg acctcgacaa tctgctggcc 
               
               
                   
               
               
                 cagattgggg atcagtacgc cgatctcttt ttggcagcaa agaacctgtc cgacgccatc 
               
               
                   
               
               
                 ctgttgagcg atatcttgag agtgaacacc gaaattacta aagcacccct tagcgcatct 
               
               
                   
               
               
                 atgatcaagc ggtacgacga gcatcatcag gatctgaccc tgctgaaggc tcttgtgagg 
               
               
                   
               
               
                 caacagctcc ccgaaaaata caaggaaatc ttctttgacc agagcaaaaa cggctacgct 
               
               
                   
               
               
                 ggctatatag atggtggggc cagtcaggag gaattctata aattcatcaa gcccattctc 
               
               
                   
               
               
                 gagaaaatgg acggcacaga ggagttgctg gtcaaactta acagggagga cctgctgcgg 
               
               
                   
               
               
                 aagcagcgga cctttgacaa cgggtctatc ccccaccaga ttcatctggg cgaactgcac 
               
               
                   
               
               
                 gcaatcctga ggaggcagga ggatttttat ccttttctta aagataaccg cgagaaaata 
               
               
                   
               
               
                 gaaaagattc ttacattcag gatcccgtac tacgtgggac ctctcgcccg gggcaattca 
               
               
                   
               
               
                 cggtttgcct ggatgacaag gaagtcagag gagactatta caccttggaa cttcgaagaa 
               
               
                   
               
               
                 gtggtggaca agggtgcatc tgcccagtct ttcatcgagc ggatgacaaa ttttgacaag 
               
               
                   
               
               
                 aacctcccta atgagaaggt gctgcccaaa cactctctgc tctacgagta ctttaccgtc 
               
               
                   
               
               
                 tacaatgaac tgactaaagt caagtacgtc accgagggaa tgaggaagcc ggcattcctt 
               
               
                   
               
               
                 agtggagaac agaagaaggc gattgtagac ctgttgttca agaccaacag gaaggtgact 
               
               
                   
               
               
                 gtgaagcaac ttaaagaaga ctactttaag aagatcgaat gttttgacag tgtggaaatt 
               
               
                   
               
               
                 tcaggggttg aagaccgctt caatgcgtca ttggggactt accatgatct tctcaagatc 
               
               
                   
               
               
                 ataaaggaca aagacttcct ggacaacgaa gaaaatgagg atattctcga agacatcgtc 
               
               
                   
               
               
                 ctcaccctga ccctgttcga agacagggaa atgatagaag agcgcttgaa aacctatgcc 
               
               
                   
               
               
                 cacctcttcg acgataaagt tatgaagcag ctgaagcgca ggagatacac aggatgggga 
               
               
                   
               
               
                 agattgtcaa ggaagctgat caatggaatt agggataaac agagtggcaa gaccatactg 
               
               
                   
               
               
                 gatttcctca aatctgatgg cttcgccaat aggaacttca tgcaactgat tcacgatgac 
               
               
                   
               
               
                 tctcttacct tcaaggagga cattcaaaag gctcaggtga gcgggcaggg agactccctt 
               
               
                   
               
               
                 catgaacaca tcgcgaattt ggcaggttcc cccgctatta aaaagggcat ccttcaaact 
               
               
                   
               
               
                 gtcaaggtgg tggatgaatt ggtcaaggta atgggcagac ataagccaga aaatattgtg 
               
               
                   
               
               
                 atcgagatgg cccgcgaaaa ccagaccaca cagaagggcc agaaaaatag tagagagcgg 
               
               
                   
               
               
                 atgaagagga tcgaggaggg catcaaagag ctgggatctc agattctcaa agaacacccc 
               
               
                   
               
               
                 gtagaaaaca cacagctgca gaacgaaaaa ttgtacttgt actatctgca gaacggcaga 
               
               
                   
               
               
                 gacatgtacg tcgaccaaga acttgatatt aatagactgt ccgactatga cgtagaccat 
               
               
                   
               
               
                 atcgtgcccc agtccttcct gaaggacgac tccattgata acaaagtctt gacaagaagc 
               
               
                   
               
               
                 gacaagaaca ggggtaaaag tgataatgtg cctagcgagg aggtggtgaa aaaaatgaag 
               
               
                   
               
               
                 aactactggc gacagctgct taatgcaaag ctcattacac aacggaagtt cgataatctg 
               
               
                   
               
               
                 acgaaagcag agagaggtgg cttgtctgag ttggacaagg cagggtttat taagcggcag 
               
               
                   
               
               
                 ctggtggaaa ctaggcagat cacaaagcac gtggcgcaga ttttggacag ccggatgaac 
               
               
                   
               
               
                 acaaaatacg acgaaaatga taaactgata cgagaggtca aagttatcac gctgaaaagc 
               
               
                   
               
               
                 aagctggtgt ccgattttcg gaaagacttc cagttctaca aagttcgcga gattaataac 
               
               
                   
               
               
                 taccatcatg ctcacgatgc gtacctgaac gctgttgtcg ggaccgcctt gataaagaag 
               
               
                   
               
               
                 tacccaaagc tggaatccga gttcgtatac ggggattaca aagtgtacga tgtgaggaaa 
               
               
                   
               
               
                 atgatagcca agtccgagca ggagattgga aaggccacag ctaagtactt cttttattct 
               
               
                   
               
               
                 aacatcatga atttttttaa gacggaaatt accctggcca acggagagat cagaaagcgg 
               
               
                   
               
               
                 ccccttatag agacaaatgg tgaaacaggt gaaatcgtct gggataaggg cagggatttc 
               
               
                   
               
               
                 gctactgtga ggaaggtgct gagtatgcca caggtaaata tcgtgaaaaa aaccgaagta 
               
               
                   
               
               
                 cagaccggag gattttccaa ggaaagcatt ttgcctaaaa gaaactcaga caagctcatc 
               
               
                   
               
               
                 gcccgcaaga aagattggga ccctaagaaa tacgggggat ttgactcacc caccgtagcc 
               
               
                   
               
               
                 tattctgtgc tggtggtagc taaggtggaa aaaggaaagt ctaagaagct gaagtccgtg 
               
               
                   
               
               
                 aaggaactct tgggaatcac tatcatggaa agatcatcct ttgaaaagaa ccctatcgat 
               
               
                   
               
               
                 ttcctggagg ctaagggtta caaggaggtc aagaaagacc tcatcattaa actgccaaaa 
               
               
                   
               
               
                 tactctctct tcgagctgga aaatggcagg aagagaatgt tggccagcgc cggagagctg 
               
               
                   
               
               
                 caaaagggaa acgagcttgc tctgccctcc aaatatgtta attttctcta tctcgcttcc 
               
               
                   
               
               
                 cactatgaaa agctgaaagg gtctcccgaa gataacgagc agaagcagct gttcgtcgaa 
               
               
                   
               
               
                 cagcacaagc actatctgga tgaaataatc gaacaaataa gcgagttcag caaaagggtt 
               
               
                   
               
               
                 atcctggcgg atgctaattt ggacaaagta ctgtctgctt ataacaagca ccgggataag 
               
               
                   
               
               
                 cctattaggg aacaagccga gaatataatt cacctcttta cactcacgaa tctcggagcc 
               
               
                   
               
               
                 cccgccgcct tcaaatactt tgatacgact atccaccgga aacggtatac cagtaccaaa 
               
               
                   
               
               
                 gaggtcctcg atgccaccct catccaccag tcaattactg gcctgtacga aacacggatc 
               
               
                   
               
               
                 gacctctctc aactgggcgg cgactag 
               
               
                   
               
               
                 codon optimized nucleic acid sequences encoding  S. aureus  Cas9 
               
               
                 SEQ ID NO: 25 
                   
               
               
                 atgaaaagga actacattct ggggctggac atcgggatta caagcgtggg gtatgggatt 
                   
               
               
                   
               
               
                 attgactatg aaacaaggga cgtgatcgac gcaggcgtca gactgttcaa ggaggccaac 
               
               
                   
               
               
                 gtggaaaaca atgagggacg gagaagcaag aggggagcca ggcgcctgaa acgacggaga 
               
               
                   
               
               
                 aggcacagaa tccagagggt gaagaaactg ctgttcgatt acaacctgct gaccgaccat 
               
               
                   
               
               
                 tctgagctga gtggaattaa tccttatgaa gccagggtga aaggcctgag tcagaagctg 
               
               
                   
               
               
                 tcagaggaag agttttccgc agctctgctg cacctggcta agcgccgagg agtgcataac 
               
               
                   
               
               
                 gtcaatgagg tggaagagga caccggcaac gagctgtcta caaaggaaca gatctcacgc 
               
               
                   
               
               
                 aatagcaaag ctctggaaga gaagtatgtc gcagagctgc agctggaacg gctgaagaaa 
               
               
                   
               
               
                 gatggcgagg tgagagggtc aattaatagg ttcaagacaa gcgactacgt caaagaagcc 
               
               
                   
               
               
                 aagcagctgc tgaaagtgca gaaggcttac caccagctgg atcagagctt catcgatact 
               
               
                   
               
               
                 tatatcgacc tgctggagac tcggagaacc tactatgagg gaccaggaga agggagcccc 
               
               
                   
               
               
                 ttcggatgga aagacatcaa ggaatggtac gagatgctga tgggacattg cacctatttt 
               
               
                   
               
               
                 ccagaagagc tgagaagcgt caagtacgct tataacgcag atctgtacaa cgccctgaat 
               
               
                   
               
               
                 gacctgaaca acctggtcat caccagggat gaaaacgaga aactggaata ctatgagaag 
               
               
                   
               
               
                 ttccagacca tcgaaaacgt gtttaagcag aagaaaaagc ctacactgaa acagattgct 
               
               
                   
               
               
                 aaggagatcc tggtcaacga agaggacatc aagggctacc gggtgacaag cactggaaaa 
               
               
                   
               
               
                 ccagagttca ccaatctgaa agtgtatcac gatattaagg acatcacagc acggaaagaa 
               
               
                   
               
               
                 atcattgaga acgccgaact gctggatcag attgctaaga tcctgactat ctaccagagc 
               
               
                   
               
               
                 tccgaggaca tccacgaaga gctgactaac ctgaacagcg agctgaccca ggaagagatc 
               
               
                   
               
               
                 gaacagatta gtaatctgaa ggggtacacc ggaacacaca acctgtccct gaaagctatc 
               
               
                   
               
               
                 aatctgattc tggatgagct gtggcataca aacgacaatc agattgcaat ctttaaccgg 
               
               
                   
               
               
                 ctgaagctgg tcccaaaaaa ggtggacctg agtcagcaga aagagatccc aaccacactg 
               
               
                   
               
               
                 gtggacgatt tcattctgtc acccgtggtc aagcggagct tcatccagag catcaaagtg 
               
               
                   
               
               
                 atcaacgcca tcatcaagaa gtacggcctg cccaatgata tcattatcga gctggctagg 
               
               
                   
               
               
                 gagaagaaca gcaaggacgc acagaagatg atcaatgaga tgcagaaacg aaaccggcag 
               
               
                   
               
               
                 accaatgaac gcattgaaga gattatccga actaccggga aagagaacgc aaagtacctg 
               
               
                   
               
               
                 attgaaaaaa tcaagctgca cgatatgcag gagggaaagt gtctgtattc tctggaggcc 
               
               
                   
               
               
                 tccccctgg aggacctgct gaacaatcca ttcaactacg aggtcgatca tattatcccc 
               
               
                   
               
               
                 agaagcgtgt ccttcgacaa ttcctttaac aacaaggtgc tggtcaagca ggaagagaac 
               
               
                   
               
               
                 tctaaaaagg gcaataggac tcctttccag tacctgtcta gttcagattc caagatctct 
               
               
                   
               
               
                 tacgaaacct ttaaaaagca cattctgaat ctggccaaag gaaagggccg catcagcaag 
               
               
                   
               
               
                 accaaaaagg agtacctgct ggaagagcgg gacatcaaca gattctccgt ccagaaggat 
               
               
                   
               
               
                 tttattaacc ggaatctggt ggacacaaga tacgctactc gcggcctgat gaatctgctg 
               
               
                   
               
               
                 cgatcctatt tccgcgtgaa caatctggat gtgaaagtca agtccatcaa cggcgggttc 
               
               
                   
               
               
                 acatcttttc tgaggcgcaa atggaagttt aaaaaggagc gcaacaaagg gtacaagcac 
               
               
                   
               
               
                 catgccgaag atgctctgat tatcgcaaat gccgacttca tctttaagga gtggaaaaag 
               
               
                   
               
               
                 ctggacaaag ccaagaaagt gatggagaac cagatgttcg aagagaagca ggccgaatct 
               
               
                   
               
               
                 atgcccgaaa tcgagacaga acaggagtac aaggagattt tcatcactcc tcaccagatc 
               
               
                   
               
               
                 aagcatatca aggatttcaa ggactacaag tactctcacc gggtggataa aaagcccaac 
               
               
                   
               
               
                 agagagctga tcaatgacac cctgtatagt acaagaaaag acgataaggg gaataccctg 
               
               
                   
               
               
                 attgtgaaca atctgaacgg actgtacgac aaagataatg acaagctgaa aaagctgatc 
               
               
                   
               
               
                 aacaaaagtc ccgagaagct gctgatgtac caccatgatc ctcagacata tcagaaactg 
               
               
                   
               
               
                 aagctgatta tggagcagta cggcgacgag aagaacccac tgtataagta ctatgaagag 
               
               
                   
               
               
                 actgggaact acctgaccaa gtatagcaaa aaggataatg gccccgtgat caagaagatc 
               
               
                   
               
               
                 aagtactatg ggaacaagct gaatgcccat ctggacatca cagacgatta ccctaacagt 
               
               
                   
               
               
                 cgcaacaagg tggtcaagct gtcactgaag ccatacagat tcgatgtcta tctggacaac 
               
               
                   
               
               
                 ggcgtgtata aatttgtgac tgtcaagaat ctggatgtca tcaaaaagga gaactactat 
               
               
                   
               
               
                 gaagtgaata gcaagtgcta cgaagaggct aaaaagctga aaaagattag caaccaggca 
               
               
                   
               
               
                 gagttcatcg cctcctttta caacaacgac ctgattaaga tcaatggcga actgtatagg 
               
               
                   
               
               
                 gtcatcgggg tgaacaatga tctgctgaac cgcattgaag tgaatatgat tgacatcact 
               
               
                   
               
               
                 taccgagagt atctggaaaa catgaatgat aagcgccccc ctcgaattat caaaacaatt 
               
               
                   
               
               
                 gcctctaaga ctcagagtat caaaaagtac tcaaccgaca ttctgggaaa cctgtatgag 
               
               
                   
               
               
                 gtgaagagca aaaagcac.cc tcagattatc aaaaagggc 
               
               
                   
               
               
                 codon optimized nucleic acid sequences encoding  S. aureus  Cas9 
               
               
                 SEQ ID NO: 26 
                   
               
               
                 atgaagcgga actacatcct gggcctggac atcggcatca ccagcgtggg ctacggcatc 
                   
               
               
                   
               
               
                 atcgactacg agacacggga cgtgatcgat gccggcgtgc ggctgttcaa agaggccaac 
               
               
                   
               
               
                 gtggaaaaca acgagggcag gcggagcaag agaggcgcca gaaggctgaa gcggcggagg 
               
               
                   
               
               
                 cggcatagaa tccagagagt gaagaagctg ctgttcgact acaacctgct gaccgaccac 
               
               
                   
               
               
                 agcgagctga gcggcatcaa cccctacgag gccagagtga agggcctgag ccagaagctg 
               
               
                   
               
               
                 agcgaggaag agttctctgc cgccctgctg cacctggcca agagaagagg cgtgcacaac 
               
               
                   
               
               
                 gtgaacgagg tggaagagga caccggcaac gagctgtcca ccaaagagca gatcagccgg 
               
               
                   
               
               
                 aacagcaagg ccctggaaga gaaatacgtg gccgaactgc agctggaacg gctgaagaaa 
               
               
                   
               
               
                 gacggcgaag tgcggggcag catcaacaga ttcaagacca gcgactacgt gaaagaagcc 
               
               
                   
               
               
                 aaacagctgc tgaaggtgca gaaggcctac caccagctgg accagagctt catcgacacc 
               
               
                   
               
               
                 tacatcgacc tgctggaaac ccggcggacc tactatgagg gacctggcga gggcagcccc 
               
               
                   
               
               
                 ttcggctgga aggacatcaa agaatggtac gagatgctga tgggccactg cacctacttc 
               
               
                   
               
               
                 cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg acctgtacaa cgccctgaac 
               
               
                   
               
               
                 gacctgaaca atctcgtgat caccagggac gagaacgaga agctggaata ttacgagaag 
               
               
                   
               
               
                 ttccagatca tcgagaacgt gttcaagcag aagaagaagc ccaccctgaa gcagatcgcc 
               
               
                   
               
               
                 aaagaaatcc tcgtgaacga agaggatatt aagggctaca gagtgaccag caccggcaag 
               
               
                   
               
               
                 ccagagttca ccaacctgaa ggtgtaccac gacatcaagg acattaccgc ccggaaagag 
               
               
                   
               
               
                 attattgaga acgccgagct gctggatcag attgccaaga tcctgaccat ctaccagagc 
               
               
                   
               
               
                 agcgaggaca tccaggaaga actgaccaat ctgaactccg agctgaccca ggaagagatc 
               
               
                   
               
               
                 gagcagatct ctaatctgaa gggctatacc ggcacccaca acctgagcct gaaggccatc 
               
               
                   
               
               
                 aacctgatcc tggacgagct gtggcacacc aacgacaacc agatcgctat cttcaaccgg 
               
               
                   
               
               
                 ctgaagctgg tgcccaagaa ggtggacctg tcccagcaga aagagatccc caccaccctg 
               
               
                   
               
               
                 gtggacgact tcatcctgag ccccgtcgtg aagagaagct tcatccagag catcaaagtg 
               
               
                   
               
               
                 atcaacgcca tcatcaagaa gtacggcctg cccaacgaca tcattatcga gctggcccgc 
               
               
                   
               
               
                 gagaagaact ccaaggacgc ccagaaaatg atcaacgaga tgcagaagcg gaaccggcag 
               
               
                   
               
               
                 accaacgagc ggatcgagga aatcatccgg accaccggca aagagaacgc caagtacctg 
               
               
                   
               
               
                 atcgagaaga tcaagctgca cgacatgcag gaaggcaagt gcctgtacag cctggaagcc 
               
               
                   
               
               
                 atccctctgg aagatctgct gaacaacccc ttcaactatg aggtggacca catcatcccc 
               
               
                   
               
               
                 agaagcgtgt ccttcgacaa cagcttcaac aacaaggtgc tcgtgaagca ggaagaaaac 
               
               
                   
               
               
                 agcaagaagg gcaaccggac cccattccag tacctgagca gcagcgacag caagatcagc 
               
               
                   
               
               
                 tacgaaacct tcaagaagca catcctgaat ctggccaagg gcaagggcag aatcagcaag 
               
               
                   
               
               
                 accaagaaag agtatctgct ggaagaacgg gacatcaaca ggttctccgt gcagaaagac 
               
               
                   
               
               
                 ttcatcaacc ggaacctggt ggataccaga tacgccacca gaggcctgat gaacctgctg 
               
               
                   
               
               
                 cggagctact tcagagtgaa caacctggac gtgaaagtga agtccatcaa tggcggcttc 
               
               
                   
               
               
                 accagctttc tgcggcggaa atggaagttt aagaaagagc ggaacaaggg gtacaagcac 
               
               
                   
               
               
                 cacgccgagg acgccctgat cattgccaac gccgatttca tcttcaaaga gtggaagaaa 
               
               
                   
               
               
                 ctggacaagg ccaaaaaagt gatggaaaac cagatgttcg aggaaaagca ggccgagagc 
               
               
                   
               
               
                 atgcccgaga tcgaaaccga gcaggagtac aaagagatct tcatcacccc ccaccagatc 
               
               
                   
               
               
                 aagcacatta aggacttcaa ggactacaag tacagccacc gggtggacaa gaagcctaat 
               
               
                   
               
               
                 agagagctga ttaacgacac cctgtactcc acccggaagg acgacaaggg caacaccctg 
               
               
                   
               
               
                 atcgtgaaca atctgaacgg cctgtacgac aaggacaatg acaagctgaa aaagctgatc 
               
               
                   
               
               
                 aacaagagcc ccgaaaagct gctgatgtac caccacgacc cccagaccta ccagaaactg 
               
               
                   
               
               
                 aagctgatta tggaacagta cggcgacgag aagaatcccc tgtacaagta ctacgaggaa 
               
               
                   
               
               
                 accgggaact acctgaccaa gtactccaaa aaggacaacg gccccgtgat caagaagatt 
               
               
                   
               
               
                 aagtattacg gcaacaaact gaacgcccat ctggacatca ccgacgacta ccccaacagc 
               
               
                   
               
               
                 agaaacaagg tcgtgaagct gtccctgaag ccctacagat tcgacgtgta cctggacaat 
               
               
                   
               
               
                 ggcgtgtaca agttcgtgac cgtgaagaat ctggatgtga tcaaaaaaga aaactactac 
               
               
                   
               
               
                 gaagtgaata gcaagtgcta tgaggaagct aagaagctga agaagatcag caaccaggcc 
               
               
                   
               
               
                 gagtttatcg cctccttcta caacaacgat ctgatcaaga tcaacggcga gctgtataga 
               
               
                   
               
               
                 gtgatcggcg tgaacaacga cctgctgaac cggatcgaag tgaacatgat cgacatcacc 
               
               
                   
               
               
                 taccgcgagt acctggaaaa catgaacgac aagaggcccc ccaggatcat taagacaatc 
               
               
                   
               
               
                 gcctccaaga cccagagcat taagaagtac agcacagaca ttctgggcaa cctgtatgaa 
               
               
                   
               
               
                 gtgaaatcta agaagcaccc tcagatcatc aaaaagggc 
               
               
                   
               
               
                 codon optimized nucleic acid sequence encoding  S. aureus  Cas9 
               
               
                 SEQ ID NO: 27 
                   
               
               
                 atgaagcgca actacatcct cggactggac atcggcatta cctccgtggg atacggcatc 
                   
               
               
                   
               
               
                 atcgattacg aaactaggga tgtgatcgac gctggagtca ggctgttcaa agaggcgaac 
               
               
                   
               
               
                 gtggagaaca acgaggggcg gcgctcaaag aggggggccc gccggctgaa gcgccgccgc 
               
               
                   
               
               
                 agacatagaa tccagcgcgt gaagaagctg ctgttcgact acaaccttct gaccgaccac 
               
               
                   
               
               
                 tccgaacttt ccggcatcaa cccatatgag gctagagtga agggattgtc ccaaaagctg 
               
               
                   
               
               
                 tccgaggaag agttctccgc cgcgttgctc cacctcgcca agcgcagggg agtgcacaat 
               
               
                   
               
               
                 gtgaacgaag tggaagaaga taccggaaac gagctgtcca ccaaggagca gatcagccgg 
               
               
                   
               
               
                 aactccaagg ccctggaaga gaaatacgtg gcggaactgc aactggagcg gctgaagaaa 
               
               
                   
               
               
                 gacggagaag tgcgcggctc gatcaaccgc ttcaagacct cggactacgt gaaggaggcc 
               
               
                   
               
               
                 aagcagctcc tgaaagtgca aaaggcctat caccaacttg accagtcctt tatcgatacc 
               
               
                   
               
               
                 tacatcgatc tgctcgagac tcggcggact tactacgagg gtccagggga gggctcccca 
               
               
                   
               
               
                 tttggttgga aggatattaa ggagtggtac gaaatgctga tgggacactg cacatacttc 
               
               
                   
               
               
                 cctgaggagc tgcggagcgc gaaatacgca tacaacgcag acctgtacaa cgcgctgaac 
               
               
                   
               
               
                 gacctgaaca atctcgtgat cacccgggac gagaacgaaa agctcgagta ttacgaaaag 
               
               
                   
               
               
                 ttccagatta ttgagaacgt gttcaaacag aagaagaagc cgacactgaa gcagattgcc 
               
               
                   
               
               
                 aaggaaatcc tcgtgaacga agaggacatc aagggctatc gagtgacctc aacgggaaag 
               
               
                   
               
               
                 ccggagttca ccaatctgaa ggtctaccac gacatcaaag acattaccgc ccggaaggag 
               
               
                   
               
               
                 atcattgaga acgcggagct gttggaccag attgcgaaga ttctgaccat ctaccaatcc 
               
               
                   
               
               
                 tccgaggata ttcaggaaga actcaccaac ctcaacagcg aactgaccca ggaggagata 
               
               
                   
               
               
                 gagcaaatct ccaacctgaa gggctacacc ggaactcata acctgagcct gaaggccatc 
               
               
                   
               
               
                 aacttgatcc tggacgagct gtggcacacc aacgataacc agatcgctat tttcaatcgg 
               
               
                   
               
               
                 ctgaagctgg tccccaagaa agtggacctc tcacaacaaa aggagatccc tactaccctt 
               
               
                   
               
               
                 gtggacgatt tcattctgtc ccccgtggtc aagagaagct tcatacagtc aatcaaagtg 
               
               
                   
               
               
                 atcaatgcca ttatcaagaa atacggtctg cccaacgaca ttatcattga gctcgcccgc 
               
               
                   
               
               
                 gagaagaact cgaaggacgc ccagaagatg attaacgaaa tgcagaagag gaaccgacag 
               
               
                   
               
               
                 actaacgaac ggatcgaaga aatcatccgg accaccggga aggaaaacgc gaagtacctg 
               
               
                   
               
               
                 atcgaaaaga tcaagctcca tgacatgcag gaaggaaagt gtctgtactc gctggaggcc 
               
               
                   
               
               
                 attccgctgg aggacttgct gaacaaccct tttaactacg aagtggatca tatcattccg 
               
               
                   
               
               
                 aggagcgtgt cattcgacaa ttccttcaac aacaaggtcc tcgtgaagca ggaggaaaac 
               
               
                   
               
               
                 tcgaagaagg gaaaccgcac gccgttccag tacctgagca gcagcgactc caagatttcc 
               
               
                   
               
               
                 tacgaaacct tcaagaagca catcctcaac ctggcaaagg ggaagggtcg catctccaag 
               
               
                   
               
               
                 accaagaagg aatatctgct ggaagaaaga gacatcaaca gattctccgt gcaaaaggac 
               
               
                   
               
               
                 ttcatcaacc gcaacctcgt ggatactaga tacgctactc ggggtctgat gaacctcctg 
               
               
                   
               
               
                 agaagctact ttagagtgaa caatctggac gtgaaggtca agtcgattaa cggaggtttc 
               
               
                   
               
               
                 acctccttcc tgcggcgcaa gtggaagttc aagaaggaac ggaacaaggg ctacaagcac 
               
               
                   
               
               
                 cacgccgagg acgccctgat cattgccaac gccgacttca tcttcaaaga atggaagaaa 
               
               
                   
               
               
                 cttgacaagg ctaagaaggt catggaaaac cagatgttcg aagaaaagca ggccgagtct 
               
               
                   
               
               
                 atgcctgaaa tcgagactga acaggagtac aaggaaatct ttattacgcc acaccagatc 
               
               
                   
               
               
                 aaacacatca aggatttcaa ggattacaag tactcacatc gcgtggacaa aaagccgaac 
               
               
                   
               
               
                 agggaactga tcaacgacac cctctactcc acccggaagg atgacaaagg gaataccctc 
               
               
                   
               
               
                 atcgtcaaca accttaacgg cctgtacgac aaggacaacg ataagctgaa gaagctcatt 
               
               
                   
               
               
                 aacaagtcgc ccgaaaagtt gctgatgtac caccacgacc ctcagactta ccagaagctc 
               
               
                   
               
               
                 aagctgacca tggagcagta tggggacgag aaaaacccgt tgtacaagta ctacgaagaa 
               
               
                   
               
               
                 actgggaatt atctgactaa gtactccaag aaagataacg gccccgtgat taagaagatt 
               
               
                   
               
               
                 aagtactacg gcaacaagct gaacgcccat ctggacatca ccgatgacta ccctaattcc 
               
               
                   
               
               
                 cgcaacaagg tcgtcaagct gagcctcaag ccctaccggt ttgatgtgta ccttgacaat 
               
               
                   
               
               
                 ggagtgtaca agttcgtgac tgtgaagaac cttgacgtga tcaagaagga gaactactac 
               
               
                   
               
               
                 gaagtcaact ccaagtgcta cgaggaagca aagaagttga agaagatctc gaaccaggcc 
               
               
                   
               
               
                 gagttcattg cctccttcta taacaacgac ctgattaaga tcaacggcga actgtaccgc 
               
               
                   
               
               
                 gtcattggcg tgaacaacga tctcctgaac cgcatcgaag tgaacatgat cgacatcact 
               
               
                   
               
               
                 taccgggaat acctggagaa tatgaacgac aagcgcccgc cccggatcat taagactatc 
               
               
                   
               
               
                 gcctcaaaga cccagtcgat caagaagtac agcaccgaca tcctgggcaa cctgtacgag 
               
               
                   
               
               
                 gtcaaatcga agaagcaccc ccagatcatc aagaaggga 
               
               
                   
               
               
                 codon optimized nucleic acid sequence encoding  S. aureus  Cas9 
               
               
                 SEQ ID NO: 28 
                   
               
               
                 atggccccaaagaagaagcggaaggtcggtatccacggagtcccagcagccaagcggaactacatcct 
                   
               
               
                   
               
               
                 gggcctggacatcggcatcaccagcgtgggctacggcatcatcgactacgagacacgggacgtgatcg 
               
               
                   
               
               
                 atgccggcgtgcggctgttcaaagaggccaacgtggaaaacaacgagggcaggcggagcaagagaggc 
               
               
                   
               
               
                 gccagaaggctgaagcggcggaggcggcatagaatccagagagtgaagaagctgctgttcgactacaa 
               
               
                   
               
               
                 cctgctgaccgaccacagcgagctgagcggcatcaacccctacgaggccagagtgaagggcctgagcc 
               
               
                   
               
               
                 agaagctgagcgaggaagagttctctgccgccctgctgcacctggccaagagaagaggcgtgcacaac 
               
               
                   
               
               
                 gtgaacgaggtggaagaggacaccggcaacgagctgtccaccagagagcagatcagccggaacagcaa 
               
               
                   
               
               
                 ggccctggaagagaaatacgtggccgaactgcagctggaacggctgaagaaagacggcgaagtgcggg 
               
               
                   
               
               
                 gcagcatcaacagattcaagaccagcgactacgtgaaagaagccaaacagctgctgaaggtgcagaag 
               
               
                   
               
               
                 gcctaccaccagctggaccagagcttcatcgacacctacatcgacctgctggaaacccggcggaccta 
               
               
                   
               
               
                 ctatgagggacctggcgagggcagccccttcggctggaaggacatcaaagaatggtacgagatgctga 
               
               
                   
               
               
                 tgggccactgcacctacttccccgaggaactgcggagcgtgaagtacgcctacaacgccgacctgtac 
               
               
                   
               
               
                 aacgccctgaacgacctgaacaatctcgtgatcaccagggacgagaacgagaagctggaatattacga 
               
               
                   
               
               
                 gaagttccagatcatcgagaacgtgttcaagcagaagaagaagcccaccctgaagcagatcgccaaag 
               
               
                   
               
               
                 aaatcctcgtgaacgaagaggatattaagggctacagagtgaccagcaccggcaagcccgagttcacc 
               
               
                   
               
               
                 aacctgaaggtgtaccacgacatcaaggacattaccgcccggaaagagattattgagaacgccgagct 
               
               
                   
               
               
                 gctggatcagattgccaagatcctgaccatctaccagagcagcgaggacatccaggaagaactgacca 
               
               
                   
               
               
                 atctgaactccgagctgacccaggaagagatcgagcagatctctaatctgaagggctataccggcacc 
               
               
                   
               
               
                 cacaacctgagcctgaaggccatcaacctgatcctggacgagctgtggcacaccaacgacaaccagat 
               
               
                   
               
               
                 cgctatcttcaaccggctgaagctggtgcccaagaaggtggacctgtcccagcagaaagagatcccca 
               
               
                   
               
               
                 ccaccctggtggacgacttcatcctgagccccgtcgtgaagagaagcttcatccagagcatcaaagtg 
               
               
                   
               
               
                 atcaacgccatcatcaagaagtacggcctgcccaacgacatcattatcgagctggcccgcgagaagaa 
               
               
                   
               
               
                 ctccaaggacgcccagaaaatgatcaacgagatgcagaagcggaaccggcagaccaacgagcggatcg 
               
               
                   
               
               
                 aggaaatcatccggaccaccggcaaagagaacgccaagtacctgatcgagaagatcaagctgcacgac 
               
               
                   
               
               
                 atgcaggaaggcaagtgcctgtacagcctggaagccatccctctggaagatctgctgaacaacccctt 
               
               
                   
               
               
                 caactatgaggtggaccacatcatccccagaagcgtgtccttcgacaacagcttcaacaacaaggtgc 
               
               
                   
               
               
                 tcgtgaagcaggaagaaaacagcaagaagggcaaccggaccccattccagtacctgagcagcagcgac 
               
               
                   
               
               
                 agcaagatcagctacgaaaccttcaagaagcacatcctgaatctggccaagggcaagggcagaatcag 
               
               
                   
               
               
                 caagaccaagaaagagtatctgctggaagaacgggacatcaacaggttctccgtgcagaaagacttca 
               
               
                   
               
               
                 tcaaccggaacctggtggataccagatacgccaccagaggcctgatgaacctgctgcggagctacttc 
               
               
                   
               
               
                 agagtgaacaacctggacgtgaaagtgaagtccatcaatggcggcttcaccagctttctgcggcggaa 
               
               
                   
               
               
                 gtggaagtttaagaaagagcggaacaaggggtacaagcaccacgccgaggacgccctgatcattgcca 
               
               
                   
               
               
                 acgccgatttcatcttcaaagactggaagaaactggacaaggccaaaaaagtgatggaaaaccagatg 
               
               
                   
               
               
                 ttcgaggaaaggcaggccgagagcatgcccgagatcgaaaccgagcaggagtacaaagagatcttcat 
               
               
                   
               
               
                 caccccccaccagatcaagcacattaaggacttcaaggactacaagtacagccaccgggtggacaaga 
               
               
                   
               
               
                 agcctaatagagagctgattaacgacaccctgtactccacccggaaggacgacaagggcaacaccctg 
               
               
                   
               
               
                 atcgtgaacaatctgaacggcctgtacgacaaggacaatgacaagctgaaaaagctgatcaacaagag 
               
               
                   
               
               
                 ccccgaaaagctgctgatgtaccaccacgacccccagacctaccagaaactgaagctgattatggaac 
               
               
                   
               
               
                 agtacggcgacgagaagaatcccctgtacaagtactacgaggaaaccgggaactacctgaccaagtac 
               
               
                   
               
               
                 tccaaaaaggacaacggccccgtgatcaagaagattaagtattacggcaacaaactgaacgcccatct 
               
               
                   
               
               
                 ggacatcaccgacgactaccccaacagcagaaacaaggtcgtgaagctgtccctgaagccctacagat 
               
               
                   
               
               
                 tcgacgtgtacctggacaatggcgtgtacaagttcgtgaccgtgaagaatctggatgtgatcaaaaaa 
               
               
                   
               
               
                 gaaaactactacgaagtgaatagcaagtgctatgaggaagctaagaagctgaagaagatcagcaacca 
               
               
                   
               
               
                 ggccgagtttatcgcctccttctacaacaacgatctgatcaagatcaacggcgagctgtatagagtga 
               
               
                   
               
               
                 tcggcgtgaacaacgacctgctgaaccggatcgaagtgaacatgatcgacatcacctaccgcgagtac 
               
               
                   
               
               
                 ctggaaaacatgaacgacaagaggccccccaggatcattaagacaatcgcctccaagacccagagcat 
               
               
                   
               
               
                 taagaagtacagcacagacattctgggcaacctgtatgaagtgaaatctaagaagcaccctcagatca 
               
               
                   
               
               
                 tcaaaaagggcaaaaggccggcggccacgaaaaaggccggccaggcaaaaaagaaaaag 
               
               
                   
               
               
                 codon optimized nucleic acid sequence encoding  S. aureus  Cas9 
               
               
                 SEQ ID NO: 29 
                   
               
               
                 accggtgcca ccatgtaccc atacgatgtt ccagattacg cttcgccgaa gaaaaagcgc 
                   
               
               
                   
               
               
                 aaggtcgaag cgtccatgaa aaggaactac attctggggc tggacatcgg gattacaagc 
               
               
                   
               
               
                 gtggggtatg ggattattga ctatgaaaca agggacgtga tcgacgcagg cgtcagactg 
               
               
                   
               
               
                 ttcaaggagg ccaacgtgga aaacaatgag ggacggagaa gcaagagggg agccaggcgc 
               
               
                   
               
               
                 ctgaaacgac ggagaaggca cagaatccag agggtgaaga aactgctgtt cgattacaac 
               
               
                   
               
               
                 ctgctgaccg accattctga gctgagtgga attaatcctt atgaagccag ggtgaaaggc 
               
               
                   
               
               
                 ctgagtcaga agctgtcaga ggaagagttt tccgcagctc tgctgcacct ggctaagcgc 
               
               
                   
               
               
                 cgaggagtgc ataacgtcaa tgaggtggaa gaggacaccg gcaacgagct gtctacaaag 
               
               
                   
               
               
                 gaacagatct cacgcaatag caaagctctg gaagagaagt atgtcgcaga gctgcagctg 
               
               
                   
               
               
                 gaacggctga agaaagatgg cgaggtgaga gggtcaatta ataggttcaa gacaagcgac 
               
               
                   
               
               
                 tacgtcaaag aagccaagca gctgctgaaa gtgcagaagg cttaccacca gctggatcag 
               
               
                   
               
               
                 agcttcatcg atacttatat cgacctgctg gagactcgga gaacctacta tgaggcacca 
               
               
                   
               
               
                 ggagaaggga gccccttcgg atggaaagac atcaaggaat ggtacgagat gctgatggga 
               
               
                   
               
               
                 cattgcacct attttccaga agagctgaga agcgtcaagt acgcttataa cgcagatct 
               
               
                   
               
               
                 tacaacgccc tgaatgacct gaacaacctg gtcatcacca gggatgaaaa cgagaaactg 
               
               
                   
               
               
                 gaatactatg agaagttcca gatcatcgaa aacgtgttta agcagaagaa aaagcctaca 
               
               
                   
               
               
                 ctgaaacaga ttgctaagga gatcctggtc aacgaagagg acatcaaggg ctaccgggtg 
               
               
                   
               
               
                 acaagcactg gaaaaccaga gttcaccaat ctgaaagtgt atcacgatat taaggacatc 
               
               
                   
               
               
                 acagcacgga aagaaatcat tgagaacgcc gaactgctgg atcagattgc taagatcctg 
               
               
                   
               
               
                 actatctacc agagctccga ggacatccag gaagagctga ctaacctgaa cagcgagctg 
               
               
                   
               
               
                 acccaggaag agatcgaaca gattagtaat ctgaaggggc acaccggaac acacaacctg 
               
               
                   
               
               
                 tccctgaaag ctatcaatct gattctggat gagctgtggc atacaaacga caatcagatt 
               
               
                   
               
               
                 gcaatcttta accggctgaa gctggtccca aaaaaggtgg acctgagtca gcagaaagag 
               
               
                   
               
               
                 atcccaacca cactggtgga cgatttcatt ctgtcacccg tggtcaagcg gagcttcatc 
               
               
                   
               
               
                 cagagcatca aagtgatcaa cgccatcatc aagaagtacg gcctgcccaa tgatatcatt 
               
               
                   
               
               
                 atcgagctgg ctagggagaa gaacagcaag gacgcacaga agatgatcaa tgagatgcag 
               
               
                   
               
               
                 aaacgaaacc ggcagaccaa tgaacgcatt gaagagatta tccgaactac cgggaaagag 
               
               
                   
               
               
                 aacgcaaagt acctgattga aaaaatcaag ctgcacgata tgcaggaggg aaagtgtctg 
               
               
                   
               
               
                 tattctctgg aggccatccc cctggaggac ctgctgaaca atccattcaa ctacgaggtc 
               
               
                   
               
               
                 gatcatatta tccccagaag cgtgtccttc gacaattcct ttaacaacaa ggtgctggtc 
               
               
                   
               
               
                 aagcaggaag agaactctaa aaagggcaat aggactcctt tccagtacct gtctagttca 
               
               
                   
               
               
                 gattccaaga tctcttacga aacctttaaa aagcacattc tgaatctggc caaaggaaag 
               
               
                   
               
               
                 ggccgcatca gcaagaccaa aaaggagtac ctgctggaag agcgggacat caacagattc 
               
               
                   
               
               
                 tccgtccaga aggattttat taaccggaat ctggtggaca caagatacgc tactcgcggc 
               
               
                   
               
               
                 ctgatgaatc tgctgcgatc ctatttccgg gtgaacaatc tggatgtgaa agtcaagtcc 
               
               
                   
               
               
                 atcaacggcg ggttcacatc ttttctgagg cgcaaatgga agtttaaaaa ggagcgcaac 
               
               
                   
               
               
                 aaagggtaca agcaccatgc cgaagatgct ctgattatcg caaatgccga cttcatcttt 
               
               
                   
               
               
                 aaggagtgga aaaagctgga caaagccaag aaagtgatgg agaaccagat gttcgaagag 
               
               
                   
               
               
                 aagcaggccg aatctatgcc cgaaatcgag acagaacagg agtacaagga gattttcatc 
               
               
                   
               
               
                 actcctcacc agatcaagca tatcaaggat ttcaaggact acaagtactc tcaccgggtg 
               
               
                   
               
               
                 gataaaaagc ccaacagaga gctgatcaat gacaccctgt atagtacaag aaaagacgat 
               
               
                   
               
               
                 aaggggaata ccctgattgt gaacaatctg aacggactgt acgacaaaga taatgacaag 
               
               
                   
               
               
                 ctgaaaaagc tgatcaacaa aagtcccgag aagctgctga tgtaccacca tgatcctcag 
               
               
                   
               
               
                 acatatcaga aactgaagct gattatggag cagtacggcg acgagaagaa cccactgtat 
               
               
                   
               
               
                 aagtactatg aagagactgg gaactacctg accaagtata gcaaaaagga taatggcccc 
               
               
                   
               
               
                 gtgatcaaga agaccaagta ctatgggaac aagctgaatg cccatctgga catcacagac 
               
               
                   
               
               
                 gattacccta acagtcgcaa caaggtggtc aagctgtcac tgaagccata cagattcgat 
               
               
                   
               
               
                 gtctatctgg acaacggcgt gtataaattt gtgactgtca agaatctgga tgtcatcaaa 
               
               
                   
               
               
                 aaggagaact actatgaagt gaatagcaag tgctacgaag aggctaaaaa gctgaaaaag 
               
               
                   
               
               
                 attagcaacc aggcagagtt catcgcctcc ttttacaaca acgacctgat taagatcaat 
               
               
                   
               
               
                 ggcgaactgt atagggtcat cggggtgaac aatgatctgc tgaaccgcat tgaagtgaat 
               
               
                   
               
               
                 atgattgaca tcacttaccg agagtatctg gaaaacatga atgataagcg cccccctcga 
               
               
                   
               
               
                 attatcaaaa caattgcctc taagactcag agtatcaaaa agtactcaac cgacattctg 
               
               
                   
               
               
                 ggaaacctgt atgaggtgaa gagcaaaaag caccctcaga ttatcaaaaa gggctaagaa 
               
               
                   
               
               
                 ttc 
               
               
                   
               
               
                 codon optimized nucleic acid sequences encoding  S. aureus  Cas9 
               
               
                 SEQ ID NO: 30 
                   
               
               
                   atggccccaaagaagaagcggaaggtcggtatccacggagtcccagcagcc aagcggaactacatcct 
                   
               
               
                   
               
               
                 gggcctggacatcggcatcaccagcgtgggctacggcatcatcgactacgagacacgggacgtgatcg 
               
               
                   
               
               
                 atgccggcgtgcggctgttcaaagaggccaacgtggaaaacaacgagggcaggcggagcaagagaggc 
               
               
                   
               
               
                 gccagaaggctgaagcggcggaggcggcatagaatccagagagtgaagaagctgctgttcgactacaa 
               
               
                   
               
               
                 cctgctgaccgaccacagcgagctgagcggcatcaacccctacgaggccagagtgaagggcctgagcc 
               
               
                   
               
               
                 agaagctgagcgaggaagagttctctgccgccctgctgcacctggccaagagaagaggcgtgcacaac 
               
               
                   
               
               
                 gtgaacgaggtggaagaggacaccggcaacgagctgtccaccaaagagcagatcagccggaacagcaa 
               
               
                   
               
               
                 ggccctggaagagaaatacgtggccgaactgcagcrggaacggctgaagaaagacggcgaagtgcggg 
               
               
                   
               
               
                 gcagcatcaacagattcaagaccagcgactacgtgaaagaagccaaacagctgctgaaggtgcagaag 
               
               
                   
               
               
                 gcctaccaccagctggaccagagcttcatcgacacctacatcgacctgctggaaacccggcggaccta 
               
               
                   
               
               
                 ctatgagggacctggcgagggcagccccttcggctggaaggacatcaaagaatggtacgagatgctga 
               
               
                   
               
               
                 tgggccactgcacctacttccccgaggaactgcggagcgtgaagtacgcctacaacgccgacctgtac 
               
               
                   
               
               
                 aacgccctgaacgacctgaacaatctcgtgatcaccagggacgagaacgagaagctggaatattacga 
               
               
                   
               
               
                 gaaattccagatcatcgagaacgtgttcaagcagaagaagaagcccaccctgaagcagatcgccaaag 
               
               
                   
               
               
                 aaatcctcgtgaacgaagaggatattaagggctacagagtgaccagcaccggcaagcccgagttcacc 
               
               
                   
               
               
                 aacctgaaggtgtaccacgacatcaaggacattaccgcccggaaagagattattgagaacgccgagct 
               
               
                   
               
               
                 gctggatcagattgccaagatcctgaccatctaccagagcagcgaggacatccaggaagaactgacca 
               
               
                   
               
               
                 atctgaactccgagctgacccaggaagagatcgagcagatctctaatctgaagggctataccggcacc 
               
               
                   
               
               
                 cacaacctgagcctgaaggccatcaacctgatcctggacgagctgtggcacaccaacgacaaccagat 
               
               
                   
               
               
                 cgctatcttcaaccggctgaagctggtgcccaagaaggtggacctgtcccagcagaaagagatcccca 
               
               
                   
               
               
                 ccaccctggtggacgacttcatcctgagccccgtcgtgaagagaagcttcatccagagcatcaaagtg 
               
               
                   
               
               
                 atcaacgccatcatcaagaagtacggcctgcccaacgacatcattatcgagctggcccgcgagaagaa 
               
               
                   
               
               
                 ctccaaggacgcccagaaaatgatcaacgagatgcagaagcggaaccggcagaccaacgagcggatcg 
               
               
                   
               
               
                 aggaaatcatccggaccaccggcaaagagaacgccaagtacctgatcgagaagatcaagctgcacgac 
               
               
                   
               
               
                 atgcaggaaggcaagtgcctgtacagcctggaagccatccctctggaagatctgctgaacaacccctt 
               
               
                   
               
               
                 caactatgaggtggaccacatcatccccagaagcgtgtccttcgacaacagcttcaacaacaaggtgc 
               
               
                   
               
               
                 tcgtgaagcaggaagaaaacagcaagaagggcaaccggaccccattccagtacctgagcagcagcgac 
               
               
                   
               
               
                 agcaagatcagctacgaaaccctcaagaagcacatcctgaatctggccaagggcaagggcagaatcag 
               
               
                   
               
               
                 caagaccaagaaagagtatctgctggaagaacgggacatcaacaggttctccgtgcagaaagacttca 
               
               
                   
               
               
                 tcaaccggaacctggtggataccagatacgccaccagaggcctgatgaacctgctgcggagctacttc 
               
               
                   
               
               
                 agagtgaacaacctggacgtgaaagtgaagtccatcaatggcggcttcaccagctttctgcggcggaa 
               
               
                   
               
               
                 gtggaagtttaagaaagagcggaacaaggggtacaagcaccacgccgaggacgccctgatcattgcca 
               
               
                   
               
               
                 acgccgatttcatcttcaaagagtggaagaaactggacaaggccaaaaaagtgatggaaaaccagatg 
               
               
                   
               
               
                 ttcgaggaaaagcaggccgagagcatgcccgagatcgaaaccgagcaggagtacaaagagatcttcat 
               
               
                   
               
               
                 caccccccaccagatcaagcacattaaggacttcaaggactacaagtacagccaccgggtggacaaga 
               
               
                   
               
               
                 agcctaatagagagctgattaacgacaccctgtactccacccggaaggacgacaagggcaacaccctg 
               
               
                   
               
               
                 atcgtgaacaatctgaacggcctgtacgacaaggacaatgacaagctgaaaaagctgatcaacaagag 
               
               
                   
               
               
                 coccgaaaagctgctgacgtaccaccacgacccccagacctacoagaaactgaagctgattatggaac 
               
               
                   
               
               
                 agtacggcgacgagaagaatcccctgtacaagtactacgaggaaaccgggaactacctgaccaagtac 
               
               
                   
               
               
                 tccaaaaaggacaacggccccgtgatcaagaagattaagtattacggcaacaaactgaacgcccatct 
               
               
                   
               
               
                 ggacatcaccgacgactaccccaacagcagaaacaaggtcgtgaagctgtccctgaagccctacagat 
               
               
                   
               
               
                 tcgacgtgtacctggacaatggcgtgtacaagttcgtgaccgtgaagaatctggatgtgatcaaaaaa 
               
               
                   
               
               
                 gaaaactactacgaagtgaatagcaagtgctatgaggaagctaagaagctgaagaagatcagcaacca 
               
               
                   
               
               
                 ggccgagtttatcgcctccttctacaacaacgatctgatcaagatcaacggcgagctgtatagagtga 
               
               
                   
               
               
                 tcggcgtgaacaacgacctgctgaaccggatcgaagtgaacatgatcgacatcacctaccgcgagtac 
               
               
                   
               
               
                 ctggaaaacatgaacgacaagaggccccccaggatcattaagacaatcgcctccaagacccagagcat 
               
               
                   
               
               
                 taagaagtacagcacagacattctgggcaacctgtatgaagtgaaatctaagaagcaccctcagatca 
               
               
                   
               
               
                 tcaaaaagggcaaaaggccggcggccacgaaaaaggccggccaggcaaaaaagaaaaag 
               
               
                   
               
               
                 codon optimized nucleic acid sequences encoding  S. aureus  Cas9 
               
               
                 SEQ ID NO: 31 
                   
               
               
                 aagcggaactacatcctgggcctggacatcggcatcaccagcgtgggctacggcatcatcgactacga 
                   
               
               
                   
               
               
                 gacacgggacgtgatcgatgccggcgtgcggctgttcaaagaggccaacgtggaaaacaacgagggca 
               
               
                   
               
               
                 ggcggagcaagagaggcgccagaaggctgaagcggcggaggcggcatagaatccagagagtgaagaag 
               
               
                   
               
               
                 ctgctgttcgactacaacctgctgaccgaccacagcgagctgagcggcatcaacccctacgaggccag 
               
               
                   
               
               
                 agtgaagggcctgagccagaagctgagcgaggaagagttctctgccgccctgctgcacctggccaaga 
               
               
                   
               
               
                 gaagaggcgtgcacaacgtgaacgaggtggaagaggacaccggcaacgagctgtccaccaaagagcag 
               
               
                   
               
               
                 atcagccggaacagcaaggccctggaagagaaatacgtggccgaactgcagctggaacggctgaagaa 
               
               
                   
               
               
                 agacggcgaagtgcggggcagcatcaacagattcaagaccagcgactacgtgaaagaagccaaacagc 
               
               
                   
               
               
                 tgctgaaggtgcagaaggcctaccaccagctggaccagagcttcatcgacacctacatcgacctgctg 
               
               
                   
               
               
                 gaaacccggcggacctactatgagggacctggcgagggcagccccttcggctggaaggacatcaaaga 
               
               
                   
               
               
                 atggtacgagatgctgatgggccactgcacctacttccccgaggaactgcggagcgtgaagtacgcct 
               
               
                   
               
               
                 acaacgccgacctgtacaacgccctgaacgacctgaacaatctcgtgatcaccagggacgagaacgag 
               
               
                   
               
               
                 aagctggaatattacgagaagttccagatcatcgagaacgtgttcaagcagaagaagaagcccaccct 
               
               
                   
               
               
                 gaagcagatcgccaaagaaatcctcgtgaacgaagaggatattaagggctacagagtgaccagcaccg 
               
               
                   
               
               
                 gcaagcccgagttcaccaacctgaaggtgtaccacgacatcaaggacattaccgcccggaaagagatt 
               
               
                   
               
               
                 attgagaacgccgagctgctggatcagattgccaagatcctgaccatctaccagagcagcgaggacat 
               
               
                   
               
               
                 ccaggaagaactgaccaatctgaactccgagctgacccaggaagagatcgagcagatctctaatctga 
               
               
                   
               
               
                 agggctataccggcacccacaacctgagcctgaaggccatcaacctgatcctggacgagctgtggcac 
               
               
                   
               
               
                 accaacgacaaccagatcgctatcttcaaccggctgaagctggtgcccaagaaggtggacctgtccca 
               
               
                   
               
               
                 gcagaaagagatccccaccaccctggtggacgacttcatcctgagccccgtcgtgaagagaagcttca 
               
               
                   
               
               
                 tccagagcatcaaagtgatcaacgccatcatcaagaagtacggcctgcccaacgacatcattatcgag 
               
               
                   
               
               
                 ctggcccgcgagaagaactccaaggacgcccagaaaatgatcaacgagatgcagaagcggaaccggca 
               
               
                   
               
               
                 gaccaacgagcggatcgaggaaatcatccggaccaccggcaaagagaacgccaagtacctgatcgaga 
               
               
                   
               
               
                 agatcaagctgcacgacatgcaggaaggcaagtgcctgtacagcctggaagccatccctctggaagat 
               
               
                   
               
               
                 ctgctgaacaaccccttcaactatgaggtggaccacatcatccccagaagcgtgtccttcgacaacag 
               
               
                   
               
               
                 cttcaacaacaaggtgctcgtgaagcaggaagaaaacagcaagaagggcaaccggaccccattccagt 
               
               
                   
               
               
                 acctgagcagcagcgacagcaagatcagctacgaaaccttcaagaagcacatcctgaatctggccaag 
               
               
                   
               
               
                 ggcaagggcagaatcagcaagaccaagaaagagtatctgctggaagaacgggacatcaacaggttctc 
               
               
                   
               
               
                 cgtgcagaaagacttcatcaaccggaacctggtggataccagatacgccaccagaggcctgatgaacc 
               
               
                   
               
               
                 tgctgcggagctacttcagagtgaacaacctggacgtgaaagtgaagtccatcaatggcggcttcacc 
               
               
                   
               
               
                 agctttctgcggcggaagtggaagtttaagaaagagcggaacaaggggtacaagcaccacgccgagga 
               
               
                   
               
               
                 cgccctgatcattgccaacgccgatttcatcttcaaagagtggaagaaactggacaaggccaaaaaag 
               
               
                   
               
               
                 tgatggaaaaccagatgttcgaggaaaagcaggccgagagcatgcccgagatcgaaaccgagcaggag 
               
               
                   
               
               
                 tacaaagagatcttcatcaccccccaccagatcaagcacattaaggacttcaaggactacaagtacag 
               
               
                   
               
               
                 ccaccgggtggacaagaagcctaatagagagctgattaacgacaccctgtactccacccggaaggacg 
               
               
                   
               
               
                 acaagggcaacaccctgatcgtgaacaatctgaacggcctgtacgacaaggacaatgacaagctgaaa 
               
               
                   
               
               
                 aagctgatcaacaagagccccgaaaagctgctgatgtaccaccacgacccccagacctaccagaaact 
               
               
                   
               
               
                 gaagctgattatggaacagtacggcgacgagaagaatcccctgtacaagtactacgaggaaaccggga 
               
               
                   
               
               
                 actacctgaccaagtactccaaaaaggacaacggccccgtgatcaagaagattaagtattacggcaac 
               
               
                   
               
               
                 aaactgaacgcccatctggacatcaccgacgactaccccaacagcagaaacaaggtcgtgaagctgtc 
               
               
                   
               
               
                 cctgaagccctacagattcgacgtgtacctggacaatggcgtgtacaagttcgtgaccgtgaagaatc 
               
               
                   
               
               
                 tggatgtaatcaaaaaagaaaactactacgaagtgaatagcaagtgctatgaggaagctaagaagctg 
               
               
                   
               
               
                 aagaagatcagcaaccaggccgagtttatcgcctccttctacaacaacgatctgatcaagatcaacgg 
               
               
                   
               
               
                 cgagctgtatagagtgatcggcgtgaacaacgacctgctgaaccggatcgaagtgaacatgatcgaca 
               
               
                   
               
               
                 tcacctaccgcgagtacctggaaaacatgaacgacaagaggccccccaggatcattaagacaatcgcc 
               
               
                   
               
               
                 tccaagacccagagcattaagaagtacagcacagacattctgggcaacctgtatgaagtgaaatctaa 
               
               
                   
               
               
                 gaagcaccctcagatcatcaaaaagggc 
               
               
                   
               
               
                 Vect or (pD0242) encoding codon optimized nucleic acid sequence 
               
               
                 encoding  S. aureus  Cas9 
               
               
                 SEQ ID NO: 32 
                   
               
               
                 ctaaattgtaagcgttaatattttgttaaaattcgcgttaaatttttgttaaatcagctcatttttta 
                   
               
               
                   
               
               
                 accaataggccgaaatcggcaaaatcccttataaatcaaaagaatagaccgagatagggttgagtgtt 
               
               
                   
               
               
                 gttccagtttggaacaagagtccactattaaagaacgtggactccaacgtcaaagggcgaaaaaccgt 
               
               
                   
               
               
                 ctatcagggcgatggcccactacgtgaaccatcaccctaatcaagttttttggggtcgaggtgccgta 
               
               
                   
               
               
                 aagcactaaatcggaaccctaaagggagcccccgatttagagcttgacggggaaagccggcgaacgtg 
               
               
                   
               
               
                 gcgagaaaggaagggaagaaagcgaaaggagcgggcgctagggcgctggcaagtgtagcggtcacgct 
               
               
                   
               
               
                 gcgcgtaaccaccacacccgccgcgcttaatgcgccgctacagggcgcgtcccattcgccattcaggc 
               
               
                   
               
               
                 tgcgcaactgttgggaagggcgatcggtgcgggcctcttcgctattacgccagctggcgaaaggggga 
               
               
                   
               
               
                 tgtgctgcaaggcgattaagttgggtaacgccagggttttcccagtcacgacgttgtaaaacgacggc 
               
               
                   
               
               
                 cagtgagcgcgcgtaatacgactcactatagggcgaattgggtacCtttaattctagtactatgcaTg 
               
               
                   
               
               
                 cgttgacattgattattgactagttattaatagtaatcaattacggggtcattagttcatagcccata 
               
               
                   
               
               
                 tatggagttccgcgttacataacttacggtaaatggcccgcctggctgaccgcccaacgacccccgcc 
               
               
                   
               
               
                 cattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgg 
               
               
                   
               
               
                 gtggagtatttacggtaaactgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccc 
               
               
                   
               
               
                 tattgacgtcaatgacggtaaatggcccgcctggcattatgcccagtacatgaccttatgggactttc 
               
               
                   
               
               
                 ctacttggcagtacatctacgtattagtcatcgctattaccatggtgatgcggttttggcagtacatc 
               
               
                   
               
               
                 aatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtcaatgggag 
               
               
                   
               
               
                 tttgttttggcaccaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaa 
               
               
                   
               
               
                 tgggcggtaggcgtgtacggtgggaggtetatataagcagagctctctggctaactaccggtgccacc 
               
               
                   
               
               
                 ATGAAAAGGAACTACATTCTGGGGCTGGACATCGGGATTACAAGCGTGGCTTATGGGATTATTGACTA 
               
               
                   
               
               
                 TGAAACAAGGGACGTGATCGACGCAGGCGTCAGACTGTTCAAGGAGGCCAACGTGGAAAACAATGAGG 
               
               
                   
               
               
                 GACGGAGAAGCAAGAGGGGAGCCAGGCGCCTGAAACGACGGAGAAGGCACAGAATCCAGAGGGTGAAG 
               
               
                   
               
               
                 AAACTGCTGTTCGATTACAACCTGCTGACCGACCATTCTGAGCTGAGTGGAATTAATCCTTATGAAGC 
               
               
                   
               
               
                 CAGGGTGAAAGGCCTGAGTCAGAAGCTGTCAGAGGAAGAGTTTTCCGCAGCTCTGCTGCACCTGGCTA 
               
               
                   
               
               
                 AGCGCCGAGGAGTGCATAACGTCAATGAGGTGGAAGAGGACACCGGCAACGAGCTGTCTACAAAGGAA 
               
               
                   
               
               
                 CAGATCTCACGCAATAGCAAAGCTCTGGAAGAGAAGTATGTCGCAGAGCTGCAGCTGGAACGGCTGAA 
               
               
                   
               
               
                 GAAAGATGGCGAGGTGAGAGGGTCAATTAATAGGTTCAAGACAAGCGACTACGTCAAAGAAGCCAAGC 
               
               
                   
               
               
                 AGCTGCTGAAAGTGCAGAAGGCTTACCACCAGCTGGATCAGAGCTTCATCGATACTTATATCGACCTG 
               
               
                   
               
               
                 CTGGAGACTCGGAGAACCTACTATGAGGGACCAGGAGAAGGGAGCCCCTTCGGATGGAAAGACATCAA 
               
               
                   
               
               
                 GGAATGGTACGAGATGCTGATGGGACATTGCACCTATTTTCCAGAAGAGCTGAGAAGCGTCAAGTACG 
               
               
                   
               
               
                 CTTATAACGCAGATCTGTACAACGCCCTGAATGACCTGAACAACCTGGTCATCACCAGGGATGAAAAC 
               
               
                   
               
               
                 GAGAAACTGGAATACTATGAGAAGTTCCAGATCATCGAAAACGTGTTTAAGCAGAAGAAAAAGCCTAC 
               
               
                   
               
               
                 ACTGAAACAGATTGCTAAGGAGATCCTGGTCAACGAAGAGGACATCAAGGGCTACCGGGTGACAAGCA 
               
               
                   
               
               
                 CTGGAAAACCAGAGTTCACCAATCTGAAAGTGTATCACGATATTAAGGACATCACAGCACGGAAAGAA 
               
               
                   
               
               
                 ATCATTGAGAACGCCGAACTGCTGGATCAGATTGCTAAGATCCTGACTATCTACCAGAGCTCCGAGGA 
               
               
                   
               
               
                 CATCCAGGAAGAGCTGACTAACCTGAACAGCGAGCTGACCCAGGAAGAGATCGAACAGATTAGTAATC 
               
               
                   
               
               
                 TGAAGGGGTACACCGGAACACACAACCTGTCCCTGAAAGCTATCAATCTGATTCTGGATGAGCTGTGG 
               
               
                   
               
               
                 CATACAAACGACAATCAGATTGCAATCTTTAACCGGCTGAAGCTGGTCCGAAAAAAGGTGGACCTGAG 
               
               
                   
               
               
                 TCAGCAGAAAGAGATCCCAACCACACTGGTGGACGATTTCATTCTGTCACCCGTGGTCAAGCGGAGCT 
               
               
                   
               
               
                 TCATCCAGAGCATCAAAGTGATCAACGCCATCATCAAGAAGTACGGCCTGCCCAATGATATCATTATC 
               
               
                   
               
               
                 GAGCTGGCTAGGGAGAAGAAGAGCAAGGACGCACAGAAGATGATCAATGAGATGCAGAAACGAAACCG 
               
               
                   
               
               
                 GCAGACGAATGAACGCATTGAAGAGATTATCCGAACTACCGGGAAAGAGAACGCAAAGTACCTGATTG 
               
               
                   
               
               
                 AAAAAATCAAGCTGCACGATATGCAGGAGGGAAAGTGTCTGTATTCTCTGGAGGCCATCCCCCTGGAG 
               
               
                   
               
               
                 GACCTGCTGAACAATCCATTCAACTACGAGGTCGATCATATTATCCCCAGAAGCGTGTCCTTCGACAA 
               
               
                   
               
               
                 TTCCTTTAACAACAAGGTGCTGGTCAAGCAGGAAGAGAACTCTAAAAAGGGCAATAGGACTCCTTTCC 
               
               
                   
               
               
                 AGTACCTGTCTAGTTCAGATTCCAAGATCTCTTACGAAACCTTTAAAAAGCACATTCTGAATCTGGCC 
               
               
                   
               
               
                 AAAGGAAAGGGCCGCATCAGCAAGACCAAAAAGGAGTACCTGCTGGAAGAGCGGGACATCAACAGATT 
               
               
                   
               
               
                 CTCCGTCCAGAAGGATTTTATTAACCGGAATCTGGTGGACACAAGATACGCTACTCGCGGCCTGATGA 
               
               
                   
               
               
                 ATCTGCTGCGATCCTATTTCCGGGTGAACAATCTGGATGTGAAAGTCAAGTCCATCAACGGCGGGTTC 
               
               
                   
               
               
                 ACATCTTTTCTGAGGCGCAAATGGAAGTTTAAAAAGGAGCGCAACAAAGGGTACAAGCACCATGCCGA 
               
               
                   
               
               
                 AGATGCTCTGATTATCGCAAATGCCGACTTCATCTTTAAGGAGTGGAAAAAGCTGGACAAAGCCAAGA 
               
               
                   
               
               
                 AAGTGATGGAGAACCAGATGTTCGAAGAGAAGCAGGCCGAATCTATGCCCGAAATCGAGACAGAACAG 
               
               
                   
               
               
                 GAGTACAAGGAGATTTTCATCACTCCTCACCAGATCAAGCATATCAAGGATTTCAAGGACTACAAGTA 
               
               
                   
               
               
                 CTCTCACCGGGTGGATAAAAAGCCCAACAGAGAGCTGATCAATGACACCCTGTATAGTACAAGAAAAG 
               
               
                   
               
               
                 ACGATAAGGGGAATACCCTGATTGTGAACAATCTGAACGGACTGTACGACAAAGATAATGACAAGCTG 
               
               
                   
               
               
                 AAAAAGCTGATCAACAAAAGTCCCGAGAAGCTGOTGATGTACCACCATGATCCTCAGACATATCAGAA 
               
               
                   
               
               
                 ACTGAAGCTGATTATGGAGCAGTACGGCGACGAGAAGAACCCACTGTATAAGTACTATGAAGAGACTG 
               
               
                   
               
               
                 GGAACTACCTGACCAAGTATAGCAAAAAGGATAATGGCCCCGTGATCAAGAAGATCAAGTACTATGGG 
               
               
                   
               
               
                 AACAAGCTGAATGCCCATCTGGACATCACAGACGATTACCCTAACAGTCGCAACAAGGTGGTCAAGCT 
               
               
                   
               
               
                 GTCACTGAAGCCATACAGATTCGATGTCTATCTGGACAACGGCGTGTATAAATTTGTGACTGTCAAGA 
               
               
                   
               
               
                 ATCTGGATGTCATCAAAAAGGAGAACTACTATGAAGTGAATAGCAAGTGCTACGAAGAGGCTAAAAAG 
               
               
                   
               
               
                 CTGAAAAAGATTAGCAACCAGGCAGAGTTCATCGCCTCCTTTTACAACAACGACCTGATTAAGATCAA 
               
               
                   
               
               
                 TGGCGAACTGTATAGGGTCATCGGGGTGAACAATGATCTGCTGAACCGCATTGAAGTGAATATGATTG 
               
               
                   
               
               
                 ACATCACTTACCGAGAGTATCTGGAAAACATGAATGATAAGCGCCCCCCTCGAATTATCAAAACAATT 
               
               
                   
               
               
                 GCCTCTAAGACTCAGAGTATCAAAAAGTACTCAACCGACATTCTGGGAAACCTGTATGAGGTGAAGAG 
               
               
                   
               
               
                 CAAAAAGCACCCTCAGATTATCAAAAAGGGCagcggaggcaagcgtcctgctgctactaagaaagctg 
               
               
                   
               
               
                 gtcaagctaagaaaaagaaaggatcctacccatacgatgttccagattacgcttaagaattcctagag 
               
               
                   
               
               
                 ctcgctgatcagcctcgactgtgccttctagttgccagccatctgttgtttgcccctccccegtgcct 
               
               
                   
               
               
                 tccttgaccctggaaggtgccactcccactgtcctttcctaataaaatgaggaaattgcatcgcattg 
               
               
                   
               
               
                 tctgagtaggtctcattctattctggggggtggggtggggcaggacagcaagggggaggattgggaag 
               
               
                   
               
               
                 agaatagcaggcatgctggggaggtagcggccgcCCgcggtggagctccagcttttgttccctttagt 
               
               
                   
               
               
                 gagggttaattgcgcgcttggcgtaatcatggtcatagctgtttcctgtgtgaaattgttatccgctc 
               
               
                   
               
               
                 acaattccacacaacatacgagccggaagcataaagtgtaaagcctggggtgcctaatgagtgagcta 
               
               
                   
               
               
                 actcacattaattgcgttgcgctcactgcccgctttccagtcgggaaacctgtcgtgccagctgcatt 
               
               
                   
               
               
                 aatgaatcggccaacgcgcggggagaggcggtttgcgtattgggcgctcttccgcttcctcgctcact 
               
               
                   
               
               
                 gactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggtt 
               
               
                   
               
               
                 atccacagaatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaacc 
               
               
                   
               
               
                 gtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacgagcatcacaaaaatcga 
               
               
                   
               
               
                 cgctcaagtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagctc 
               
               
                   
               
               
                 cctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaa 
               
               
                   
               
               
                 gcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctg 
               
               
                   
               
               
                 ggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtc 
               
               
                   
               
               
                 caacccggtaagacacgacttatcgccactggcagcagccactggtaacaggattagcagagcgaggt 
               
               
                   
               
               
                 atgtaggcggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtattt 
               
               
                   
               
               
                 ggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaaaca 
               
               
                   
               
               
                 aaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctc 
               
               
                   
               
               
                 aagaagatcctttgatcctttctacggggtctgacgctcagtggaacgaaaactcacgttaagggatt 
               
               
                   
               
               
                 ttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaatgaagttttaaatc 
               
               
                   
               
               
                 aatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatct 
               
               
                   
               
               
                 cagcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgg 
               
               
                   
               
               
                 gagggcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccagattt 
               
               
                   
               
               
                 atcagcaataaaccagccagccggaagggccgagcgcagaagtggtcctgcaactttatccgcctcca 
               
               
                   
               
               
                 tccagtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgcaacgtt 
               
               
                   
               
               
                 gttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatggcttcattcagctccggttc 
               
               
                   
               
               
                 ccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctc 
               
               
                   
               
               
                 cgatcgttgtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattct 
               
               
                   
               
               
                 cttactgtcatgccatccgtaagatgcttttctgtgactggtgagtactcaaccaagtcattctgaga 
               
               
                   
               
               
                 atagtgtatgcggcgaccgagttgctcttgcccggcgtcaatacgggataataccgcgccacatagca 
               
               
                   
               
               
                 gaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaaggatcttaccgctg 
               
               
                   
               
               
                 ttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccag 
               
               
                   
               
               
                 cgtttctgggtgagcaaaaacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaat 
               
               
                   
               
               
                 gttgaatactcatactcttcctttttcaatattattgaagcatttatcagggttattgtotcatgagc 
               
               
                   
               
               
                 ggatacatatttgaatgtatttagaaaaataaacaaataggggttccgcgcacatttccccgaaaagt 
               
               
                   
               
               
                 gccac 
               
               
                   
               
               
                 Human p300 
               
               
                 (with L553M mutation) protein 
               
               
                 SEQ ID NO: 33 
                   
               
               
                 MAENVVEPGPPSAKRPKLSSPALSASASDGTDFGSLFDLEHDLPDELINSTELGLTNGGDINQLQTSL 
                   
               
               
                   
               
               
                 GMVQDAASKHKQLSELLRSGSSPNLNMGVGGPGQVMASQAQQSSPGLGLINSMVKSPMTQAGLTSPNM 
               
               
                   
               
               
                 GMGTSGPNQGPTQSTGMMN8PVNQPAMGMNTGMNAGMNPGMLAAGNGQGIMPNQVMNGSIGAGRGRQK 
               
               
                   
               
               
                 MQYPNPGMGSAGNLLTEPLQQGSPQMGGQTGLRGPQPLKMGMMNNPNPYGSPYTQNPGQQIGASGLGL 
               
               
                   
               
               
                 QTQTKTVLSNNLSPFAMDKKAVPGGGMPNMGQQPAPQVQQPGLVTPVAQGMGSGAHTADPEKRKLIQQ 
               
               
                   
               
               
                 QLVLLLHAHKCQRREQANGEVRQCNLPHCRTMKNVLNHMTHCQSGKSCQVAHCASSRQIISHWKNCTR 
               
               
                   
               
               
                 KDCPVCLPLKNAGDKRNQQPILTGAPVGLGNPSSLGVGQQSAPKLSTVSQIDPSSIERAYAALGLPYQ 
               
               
                   
               
               
                 VNQMPTQPQVOAKNQQNQQPGQSPQGMRPMSNMSASPMGVNGGVGVQTPSLLSDSMLHSAINSQNPMM 
               
               
                   
               
               
                 SENASVPSMGPMPTAAQPSTTGIRKQWHEDITQDLRNHLVHKLVQAIFPTPDPAALKDRRMENLVAYA 
               
               
                   
               
               
                 RKVEGDMYESANNRAEYYHLLAEKIYKIQKELEEKRRTRLQKQNMLPNAAGMVPVSMNPGPNMGQPQP 
               
               
                   
               
               
                 GMTSNGPLPDPSMIRGSVPNQMMPRITPQSGLNQFGQMSMAQPPIVPRQTPPLQHHGQLAQPGALNPP 
               
               
                   
               
               
                 MGYGPRMQQPSNQGQFLPQTQFPSQGMNVTNIPLAPSSGQAPVSGAQMSSSSCPVNSPIMPPGSQGSH 
               
               
                   
               
               
                 IHCPQLPQPALHQNSPSPVPSRTPTPHHTPPSIGAQQPPATTIPAPVPTPPAMPPGPQSOALHPPPRQ 
               
               
                   
               
               
                 TPTPPTTQLPQQVQPSLPAAPSADQPQQQPRSQQSTAASVPTPTAPLLPPQPATPLSQPAVSIEGQVS 
               
               
                   
               
               
                 NPPSTSSTEVNSQAIAEKQPSQEVKMEAKMEVDQPEPADTQPEDISESKVEDCKMESTETEERSTELK 
               
               
                   
               
               
                 TEIKEEEDQPSTSATQSSPAPGQSKKKIFKPEELRQALMPTLEALYRQDPSSLPFRQPVDPQLLGIPD 
               
               
                   
               
               
                 YEDIVKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPV 
               
               
                   
               
               
                 MQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQKRYHFCEKCFNEIQGESVSLGDDPSQPQT 
               
               
                   
               
               
                 TINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAGFVCDGCLKKSARTRKENKFSAKR 
               
               
                   
               
               
                 LPSTRLGTFLENRVNDFLRRQNKPESGEVTVRVVHASDKTVEVKPGMKARFVDSGEMAESFPYRTKAL 
               
               
                   
               
               
                 FAFEEIDGVDLCFFGMHVQEYGSDCPPPNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKL 
               
               
                   
               
               
                 GYTTGHIWACPPSEGDDYIFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLT 
               
               
                   
               
               
                 SAKELPYFEGDFWPNVLEESIKELEOEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS 
               
               
                   
               
               
                 RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDLMDGRDAFLT 
               
               
                   
               
               
                 LARDKHLEFSSLRRAQWSTMCMLVELHTQSQDRFVYTCNECKHHVETRWHCTVCEDYDLCITCYNTKN 
               
               
                   
               
               
                 HDHKMEKLGLGLDDESNNQQAAATQSPGDSRRLSIQRCIQSLVHACQCRNANCSLPSCQKMKRVVQHT 
               
               
                   
               
               
                 KGCKRKTNGGCPICKQLIALCCYHAKHCQENKCPVPFCLNIKQKLRQQQLQHRLQQAQMLRRRMASMQ 
               
               
                   
               
               
                 RTGVVGQQQGLPSPTPATPTTPTGQQPTTPQTPQPTSQPQPTPPNSMPPYLPRTQAAGPVSQGKAAGQ 
               
               
                   
               
               
                 VTPPTPPQTAQPPLPGPPPAAVEMAMQIQRAAETQRQMAHVQIFQRPIQHQMPPMTPMAPMGMNPPPM 
               
               
                   
               
               
                 TRGPSGHLEPGMGPTGMQQQPPWSQGGLPQPQQLQSGMPRPAMMSVAQHGQPLNMAPQPGLGQVGISP 
               
               
                   
               
               
                 LKPGTVSQQALQNLLRTLRSPSSPLQQQQVLSILHANPQLLAAFTKQRAAKYANSNPQPIPGQPGMPQ 
               
               
                   
               
               
                 GQPGLQPPTMPGQQGVHSNPAMQNMNPMQAGVQRAGLPQQQPQQQLQPPMGGMSPQAQQMNMNHNTMP 
               
               
                   
               
               
                 SQFRDILRRQQMMQQQQQQGAGPGIGPGMANHNQFQQPQGVGYPPQQQQRMQHHMQQMQQGNMGQIGQ 
               
               
                   
               
               
                 LPQALGAEAGASLQAYQQRLLQQQMGSPVQPNPMSPQQHMLPNQAQSPHLQGQQIPNSLSNQVRSPQP 
               
               
                   
               
               
                 VPSPRPQSQPPHSSPSPRMQPQPSPHHVSPQTSSPHPGLVAAQANPMEQGHFASPDQNSMLSQLASNP 
               
               
                   
               
               
                 GMANLHGASATDLGLSTDNSDLNSNLSQSTLDIH 
               
               
                   
               
               
                 Human p300 Core Effect or protein 
               
               
                 (aa 1048-1664 of SEQ ID NO: 33) 
               
               
                 SEQ ID NO: 34 
                   
               
               
                 IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDIVKSPMDLSTIKRKLDTGQYQEPW 
                   
               
               
                   
               
               
                 QYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQEIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLC 
               
               
                   
               
               
                 TIPRDATYYSYONRYHFCEKCFNEIOGESVSLGDDPSQPOTTINKEOFSKRKNDTLDPELFVECTECG 
               
               
                   
               
               
                 RKMHOICVLHHEITWPAGFVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRONHPESG 
               
               
                   
               
               
                 EVTVRVVHASDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP 
               
               
                   
               
               
                 PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDYIFHCHPPDQ 
               
               
                   
               
               
                 KIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFEGDFWPNVLEESIKELEQE 
               
               
                   
               
               
                 EEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLSRGNKKKPGMPNVSNDLSQKLYATMEKH 
               
               
                   
               
               
                 KEVFFVTRLIAGPAANSLPPIVDPDPLIPCDLMDGRDAFLTIARDKHLEFSSLRRAQWSTMCMLVELH 
               
               
                   
               
               
                 TQSQF 
               
               
                   
               
               
                 VP64-dCas9-VP64 protein 
               
               
                 (with dCas9 underlined) 
               
               
                 SEQ ID NO: 35 
                   
               
               
                 RADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMVNPKKKRKVGRG MDKKY   
                   
               
               
                   
               
               
                 
                   SIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT 
                 
               
               
                   
               
               
                 
                   RRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKK 
                 
               
               
                   
               
               
                 
                   LVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAK 
                 
               
               
                   
               
               
                 
                   AILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDN 
                 
               
               
                   
               
               
                 
                   LLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPE 
                 
               
               
                   
               
               
                 
                   KYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQ 
                 
               
               
                   
               
               
                 
                   IHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEV 
                 
               
               
                   
               
               
                 
                   VDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIV 
                 
               
               
                   
               
               
                 
                   DLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILE 
                 
               
               
                   
               
               
                 
                   DIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKS 
                 
               
               
                   
               
               
                 
                   DGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR 
                 
               
               
                   
               
               
                 
                   HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRD 
                 
               
               
                   
               
               
                 
                   MYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNA 
                 
               
               
                   
               
               
                 
                   KLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVIT 
                 
               
               
                   
               
               
                 
                   LKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKS 
                 
               
               
                   
               
               
                 
                   EQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVN 
                 
               
               
                   
               
               
                 
                   IVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVK 
                 
               
               
                   
               
               
                 
                   ELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP 
                 
               
               
                   
               
               
                 
                   SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKH 
                 
               
               
                   
               
               
                 
                   RDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQL 
                 
               
               
                   
               
               
                   GGD SRADPKKKRKVASRADALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDML 
               
               
                   
               
               
                 I 
               
               
                   
               
               
                 VP64-dCas9-VP64 DNA 
               
               
                 SEQ ID NO: 36 
                   
               
               
                 cgggctgacgcattggacgattttgatctggatatgctgggaagtgacgccctcgatgattttgacct 
                   
               
               
                   
               
               
                 tgacatgcttggttcggatgcccttgatgactttgacctcgacatgctcggcagtgacgcccttgatg 
               
               
                   
               
               
                 atttcgacctggacatggttaaccccaagaagaagaggaaggtgggccgcggaatggacaagaagtac 
               
               
                   
               
               
                 tccattgggctcgccatcggcacaaacagcgtcggctgggccgtcattacggacgagtacaaggtgcc 
               
               
                   
               
               
                 gagcaaaaaattcaaagttctgggcaataccgatcgccacagcataaagaagaacctcattggcgccc 
               
               
                   
               
               
                 tcctgttcgactccggggaaaccgccgaagccacgcggctcaaaagaacagcacggcgcagatatacc 
               
               
                   
               
               
                 cgcagaaagaatcggatctga;acctgcaggagatctttagtaatgagatggctaaggtggatgactc 
               
               
                   
               
               
                 tttcttccataggctggaggagtcctttttggtggaggaggataaaaagcacgagcgccacccaatct 
               
               
                   
               
               
                 ttggcaatatcgtggacgaggtggcgtaccatgaaaagtacccaaccatatatcatctgaggaagaag 
               
               
                   
               
               
                 cttgtagacagtactgataaggctgacttgcggttgatctatctcgcgctggcgcatatgatcaaatt 
               
               
                   
               
               
                 tcggggacacttcctcatcgagggggacctgaacccagacaacagcgatgtcgacaaactctttatcc 
               
               
                   
               
               
                 aactggttcagacttacaatcagcttttcgaagagaacccgatcaacgcatccggagttgacgccaaa 
               
               
                   
               
               
                 gcaatcctgagcgctaggctgtccaaatcccggcggctcgaaaacctcatcgcacagctccctgggga 
               
               
                   
               
               
                 gaagaagaacggcctgtttggtaatcttatcgccctgtcactcgggctgacccccaactttaaatcta 
               
               
                   
               
               
                 acttcgacctggccgaagatgccaagcttcaactgagcaaagacacctacgatgatgatctcgacaat 
               
               
                   
               
               
                 ctgctggcccagatcggcgaccagtacgcagacctttttttggcggcaaagaacctgtcagacgccat 
               
               
                   
               
               
                 tctgctgagtgatattctgcgagtgaacacggagatcaccaaagctccgctgagcgctagtatgatca 
               
               
                   
               
               
                 agcgctatgatgagcaccaccaagacttgactttgctgaaggcccttgtcagacagcaactgcctgag 
               
               
                   
               
               
                 aagtacaaggaaattttcttcgatcagtctaaaaatggctacgccggatacattgacggcggagcaag 
               
               
                   
               
               
                 ccaggaggaattttacaaatttattaagcccatcttggaaaaaatggacggcaccgaggagctgctgg 
               
               
                   
               
               
                 taaagcttaacagagaagatctgttgcgcaaacagcgcactttcgacaatggaagcatcccccaccag 
               
               
                   
               
               
                 attcacctgggcgaactgcacgctatcctcaggcggcaagaggatttctacccctttttgaaagataa 
               
               
                   
               
               
                 cagggaaaagattgagaaaatcctcacatttcggataccctactatgtaggccccctcgcccggggaa 
               
               
                   
               
               
                 attccagattcgcgtggatgactcgcaaatcagaagagaccatcactccctggaacttcgaggaagtc 
               
               
                   
               
               
                 gtggataagggggcctctgcccagtccttcatcgaaaggatgactaactttgataaaaatctgcctaa 
               
               
                   
               
               
                 cgaaaaggtgcttcctaaacactctctgctgtacgagtacttcacagtttataacgagctcaccaagg 
               
               
                   
               
               
                 tcaaatacgtcacagaagggatgagaaagccagcattcctgtctggagagcagaagaaagctatcgtg 
               
               
                   
               
               
                 gacctcctcttcaagacgaaccggaaagttaccgtgaaacagctcaaagaagactatttcaaaaagat 
               
               
                   
               
               
                 tgaatgtttcgactctgttgaaatcagcggagtggaggatcgcttcaacgcatccctgggaacgtatc 
               
               
                   
               
               
                 acgatctcctgaaaatcattaaagacaaggacttcctggacaatgaggagaacgaggacattcttgag 
               
               
                   
               
               
                 gacattgtcctcacccttacgttgtttgaagatagggagatgattgaagaacgcttgaaaacttacgc 
               
               
                   
               
               
                 tcatctcttcgacgacaaagtcatgaaacagctcaagaggcgccgatatacaggatgggggcggctgt 
               
               
                   
               
               
                 caagaaaactgatcaatgggatccgagacaagcagagtggaaagacaatcctggattttcttaagtcc 
               
               
                   
               
               
                 gatggatttgccaaccggaacttcatgcagttgatccatgatgactctctcacctttaaggaggacat 
               
               
                   
               
               
                 ccagaaagcacaagtttctggccagggggacagtcttcacgagcacatcgctaatcttgcaggtagcc 
               
               
                   
               
               
                 cagctatcaaaaagggaatactgcagaccgttaaggtcgtggatgaactcgtcaaagtaatgggaagg 
               
               
                   
               
               
                 cataagcccgagaatatcgttatcgagatggcccgagagaaccaaactacccagaagggacagaagaa 
               
               
                   
               
               
                 cagtagggaaaggatgaagaggattgaagagggtataaaagaactggggtcccaaatccttaaggaac 
               
               
                   
               
               
                 acccagttgaaaacacccagcttcagaatgagaagctctacctgtactacctgcagaacggcagggac 
               
               
                   
               
               
                 atgtacgtggatcaggaactggacatcaatcggctctccgactacgacgtggatgccatcgtgcccca 
               
               
                   
               
               
                 gtcttttctcaaagatgattctattgataataaagtgttgacaagatccgataaaaatagagggaaga 
               
               
                   
               
               
                 gtgataacgtcccctcagaagaagttgtcaagaaaatgaaaaattattggcggcagctgctgaacgcc 
               
               
                   
               
               
                 aaactgatcacacaacggaagttcgataatctgactaaggctgaacgaggtggcctgtctgagttgga 
               
               
                   
               
               
                 taaagccggcctcatcaaaaggcagcttgttgagacacgccagatcaccaagcacgtggcccaaattc 
               
               
                   
               
               
                 tcgattcacgcatgaacaccaagtacgatgaaaatgacaaactgattcgagaggtgaaagttattact 
               
               
                   
               
               
                 ctgaagtctaagctggtctcagatttcagaaaggactttcagttttataaggtgagagagatcaacaa 
               
               
                   
               
               
                 ttaccaccatgcgcatgatgcctacctgaatgcagtggtaggcactgcacttatcaaaaaatatccca 
               
               
                   
               
               
                 agcttgaatctgaatttgtttacggagactataaagtgtacgatgttaggaaaatgatcgcaaagtct 
               
               
                   
               
               
                 gagcaggaaataggcaaggccaccgctaagtacttcttttacagcaatattatgaattttttcaagac 
               
               
                   
               
               
                 cgagattacactggccaatggagagattcggaagcgaccacttatcgaaacaaacggagaaacaggag 
               
               
                   
               
               
                 aaatcgtgtgggacaagggtagggatttcgcgacagtccggaaggtcctgtccatgccgcaggtgaac 
               
               
                   
               
               
                 atcgttaaaaagaccgaagtacagaccggaggcttctccaaggaaagtatcctcccgaaaaggaacag 
               
               
                   
               
               
                 cgacaagctgatcgcacgcaaaaaagattgggaccccaagaaatacggcggattcgattctcctacag 
               
               
                   
               
               
                 tcgcttacagtgtactggttgtggccaaagtggagaaagggaagtctaaaaaactcaaaagcgtcaag 
               
               
                   
               
               
                 gaactgctgggcatcacaatcatggagcgatcaagcttcgaaaaaaaccccatcgactttctcgaggc 
               
               
                   
               
               
                 gaaaggatataaagaggtcaaaaaagacctcatcattaagcttcccaagtactctctctttgagcttg 
               
               
                   
               
               
                 aaaacggccggaaacgaatgctcgctsgtgcgggcgagctgcagaaaggtaacgsgctggcactgccc 
               
               
                   
               
               
                 tctaaatacgttaatttcttgtatctggccagccactatgaaaagctcaaagggtctcccgaagataa 
               
               
                   
               
               
                 tgagcagaagcagctgttcgtggaacaacacaaacactaccttgatgagatcatcgagcaaataagcg 
               
               
                   
               
               
                 aattctccaaaagagtgatcctcgccgacgctaacctcgataaggtgctttctgcttacaataagcac 
               
               
                   
               
               
                 agggataagcccatcagggagcaggcagaaaacattatccacttgtttactctgaccaacttgggcgc 
               
               
                   
               
               
                 gcctgcagccttcaagtacttcgacaccaccatagacagaaagcggtacacctctacaaaggaggtcc 
               
               
                   
               
               
                 tggacgccacactgattcatcagtcaattacggggctctatgaaacaagaatcgacctctctcagctc 
               
               
                   
               
               
                 ggtggagacagcagggctgaccccaagaagaagaggaaggtggctagccgcgccgacgcgctggacga 
               
               
                   
               
               
                 tttcgatctcgacatgctgggttctgatgccctcgatgactttgacctggatatgttgggaagcgacg 
               
               
                   
               
               
                 cattggatgactttgatctggacatgctcggctccgatgctctggacgatttcgatctcgatatgtta 
               
               
                   
               
               
                 atc 
               
               
                   
               
               
                 Protein sequence f or SS18 
               
               
                 SEQ ID NO: 37 
                   
               
               
                 MSVAFAAPRQRGKGEITPAAIQKMLDDNNHLIQCIMDSQNKGKTSECSQYQQMLHTNLVYLATIADSN 
                   
               
               
                   
               
               
                 QNMQSLLPAPPTQNMPMGPGGMNQSGPPPPPRSHNMPSDGMVGGGPPAPHMQNQMNGQMPGPNHMPMQ 
               
               
                   
               
               
                 GPGPNOLNMTNSSMNMPSSSHGSMGGYNHSVPSSOSMPVONOMTMSQGOPMGNYGPRPNMSMOPNOGP 
               
               
                   
               
               
                 MMHQQPPSQQYNMPQGGGQHYQGQQPPMGMMGQVNQGNHMMGQRQIPPYRPPQQGPPQQYSGQEDYYG 
               
               
                   
               
               
                 DQYSHGGQGPPEGMNQQYYPDGNSQYGQQQDAYQGPPPQQGYPPQQQQYPGQQGYPGQQQGYGPSQGG 
               
               
                   
               
               
                 PGPQYPNYPQGQGQQYGGYRPTQPGPPQPPQQRPYGYDQGQYGKYQQ 
               
               
                   
               
               
                 DNA sequence f or SS18 
               
               
                 SEQ ID NO: 38 
                   
               
               
                 atgtctgtggctttcgcggccccgaggcagcgaggcaagggggagatcactcccgctgcgattcagaa 
                   
               
               
                   
               
               
                 gatgttggatgacaataaccatcttattcagtgtataatggactctcagaataaaggaaagacctcag 
               
               
                   
               
               
                 agtgttctcagtatcagcagatgttgcacacaaacttggtataccttgctacaatagcagattctaat 
               
               
                   
               
               
                 caaaatatgcagtctcttttaccagcaccacccacacagaatatgcctatgggtcctggagggatgaa 
               
               
                   
               
               
                 tcagagcggccctcccccacctccacgctctcacaacatgccttcagatggaatggtaggtgggggtc 
               
               
                   
               
               
                 ctcctgcaccgcacatgcagaaccagatgaacggccagatgcctgggcctaaccatatgcctatgcag 
               
               
                   
               
               
                 ggacctggacccaatcaactcaatatgacaaacagttccatgaatatgccttcaagtagccatggatc 
               
               
                   
               
               
                 catgggaggttacaaccattctgtgccatcatcacagagcatgccagtacagaatcagatgacaatga 
               
               
                   
               
               
                 gtcagggacaaccaatgggaaaetatggtcccagaccaaatataagtatgcagccaaaccaaggtcca 
               
               
                   
               
               
                 atgatgcatcagcagcctccttctcagcaatacaatatgccacagggaggcggacagcattaccaagg 
               
               
                   
               
               
                 acagcagccacctatgggaatgatgggtcaagttaaccaaggcaatcatatgatgggtcagagacaga 
               
               
                   
               
               
                 ttcctccctatagacctcctcaacagggcccaccacagcagtactcaggccaggaagactattacggg 
               
               
                   
               
               
                 gaccaatacagtcatggtggacaaggtcctccagaaggcatgaaccagcaatattaccctgatggaaa 
               
               
                   
               
               
                 ttcacagtatggccaacagcaagatgcataccagggaccacctccacaacagggatatccaccccagc 
               
               
                   
               
               
                 agcagcagtacccagggcagcaaggttacecaggacagcagcagggctacggtccttcacagggtggt 
               
               
                   
               
               
                 ccaggtcctcagtatcctaactacccacagggacaaggtcagcagtatggaggatatagaccaacaca 
               
               
                   
               
               
                 gcctggaccaccacagccaccccagcagaggccttatggatatgaccagggacagtatggaaattacc 
               
               
                   
               
               
                 agcag 
               
               
                   
               
               
                 Protein sequence f or VPH 
               
               
                 SEQ ID NO: 39 
                   
               
               
                 DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSLPSASVEFEGSGGPSG 
                   
               
               
                   
               
               
                 QISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTPGPPQSLSAPVPKSTQAGEGTLSEALL 
               
               
                   
               
               
                 HLQFDADEDLGALLGNSTDPGVFTDLASVDNSEFQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQ 
               
               
                   
               
               
                 RPPDpAPTPLGTSGLPNGLSGDEDFSSIADMDFSALLSQISSSGQGGGGSGFSVDTSALLDLFSPSVT 
               
               
                   
               
               
                 VPDNSLPDLDSSLASIQELLSPQEPPRPPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVL 
               
               
                   
               
               
                 FELGESSYFSEGDGFAEDPTISLLTGSEPPKAKDPTVS 
               
               
                   
               
               
                 DNA sequence f or VPH 
               
               
                 SEQ ID NO: 40 
                   
               
               
                 Gatgctttagacgattttgacttagatatgcttggttcagacgcgttagacgacttcgacctagacat 
                   
               
               
                   
               
               
                 gttaggctcagatgcattggacgacttcgatttagatatgttgggctccgatgccctagatgactttg 
               
               
                   
               
               
                 atctagatatgctagggtcactacccagcgccagcgtcgagttcgaaggcagcggcgggccttcaggg 
               
               
                   
               
               
                 cagatcagcaaccaggccctggctctggcccctagctccgctccagtgctggcccagactatggtgcc 
               
               
                   
               
               
                 ctctagtgctatggtgcctctggcccagccacctgctccagcccctgtgctgaccccaggaccacccc 
               
               
                   
               
               
                 agtcactgagcgccccagtgcccaagtctacacaggccggcgaggggactctgagtgaagctctgctg 
               
               
                   
               
               
                 cacctgcagttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccggagtgtt 
               
               
                   
               
               
                 cacagatctggcctccgtggacaactctgagtttcagcagctgctgaatcagagcgtgtccatgtctc 
               
               
                   
               
               
                 atagtacagccgaaccaatgctgatggagtaccccgaagccattacccggctggtgaccggcagccag 
               
               
                   
               
               
                 cggccccccgaccccgctccaactcccctgggaaccagcggcctgcctaatgggctgtccggagatga 
               
               
                   
               
               
                 agacttctcaagcatcgctgatatggactttagtgccctgctgtcacagatttcctctagtgggcagg 
               
               
                   
               
               
                 gaggaggtggaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcccctcggtgacc 
               
               
                   
               
               
                 gtgcccgacatgagcctgcctgaccttgacagcagcctggccagtatccaagagctcctgtctcccca 
               
               
                   
               
               
                 ggagccccccaggcctcccgaggcagagaacagcagcccggattcagggaagcagctggtgcactaca 
               
               
                   
               
               
                 cagcgcagccgctgttcctgctggaccccggctccgtggacaccgggagcaacgacctgccggtgctg 
               
               
                   
               
               
                 tttgagctgggagagggctcctacttctccgaaggggacggcttcgccgaggaccccaccatctccct 
               
               
                   
               
               
                 gctgacaggctcggagcctcccaaagccaaggaccccactgtctcc 
               
               
                   
               
               
                 Protein sequence f or VPR 
               
               
                 SEQ ID NO: 41 
                   
               
               
                 DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSPKKKRKVGSQYLPDTD 
                   
               
               
                   
               
               
                 DRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPOPYPFTSSLSTINYD 
               
               
                   
               
               
                 EFPTMVFPSGOISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPT 
               
               
                   
               
               
                 QAGEGTLSEALLOLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQOLLNQGIPVAPHTTEPMLMEYP 
               
               
                   
               
               
                 EAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISSG5GSGSRD5REGMF 
               
               
                   
               
               
                 LPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPL 
               
               
                   
               
               
                 DPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLES 
               
               
                   
               
               
                 MTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF 
               
               
                   
               
               
                 DNA sequence f or VPR 
               
               
                 SEQ ID NO: 42 
                   
               
               
                 gatgctttagacgattttgacttagatatgcttggttcagacgcgttagacgacttcgacctagacat 
                   
               
               
                   
               
               
                 gttaggctcagatgcattggacgacttcgatttagatatgttgggctccgatgccctagatgactttg 
               
               
                   
               
               
                 atctagatatgctaggtagtcccaaaaagaagaggaaagtgggatcccagtatctgcccgacacagat 
               
               
                   
               
               
                 gatagacaccgaatcgaagagaaacgcaagcgaacgtatgaaaccttcaaatcgatcatgaagaaatc 
               
               
                   
               
               
                 gcccttctcgggtccgaccgatcccaggcccccaccgagaaggattgcggtcccgtcccgctcgtcgg 
               
               
                   
               
               
                 ccagcgtgccgaagcctgcgccgcagccctaccccttcacgtcgagcctgagcacaatcaattatgac 
               
               
                   
               
               
                 gagttcccgacgatggtgttcccctcgggacaaatctcacaagcctcggcgctcgcaccagcgcctcc 
               
               
                   
               
               
                 ccaagtccttccgcaagcgcctgccccagcgcctgcaccggcaatggtgtccgccctcgcacaggccc 
               
               
                   
               
               
                 ctgcgcccgtccccgtgctcgcgcctggaccgccccaggcggtcgctccaccggctccgaagccgacg 
               
               
                   
               
               
                 caggccggagagggaacactctccgaagcacttcttcaactccagtttgatgacgaggatcttggagc 
               
               
                   
               
               
                 actccttggaaactcgacagaccctgcggtgtttaccgacctcgcgtcagtagataactccgaatttc 
               
               
                   
               
               
                 agcagcttttgaaccagggtatcccggtcgcgccacatacaacggagcccatgttgatggaatacccc 
               
               
                   
               
               
                 gaagcaatcacgagacttgtgacgggagcgcagcggcctcccgatcccgcacccgcacctttgggggc 
               
               
                   
               
               
                 acctggcctccctaacggacttttgagcggcgacgaggatttctcctccatcgccgatatggatttct 
               
               
                   
               
               
                 cagccttgctgtcacagatttccagcggctctggcagcggcagccgggattccagggaagggatgttt 
               
               
                   
               
               
                 ttgccgaagcctgaggccggctccgctattagtgacgtgtttgagggccgcgaggtgtgccagccaaa 
               
               
                   
               
               
                 acgaatccggccatttcatcctccaggaagtccatgggccaaccgcccactccccgccagcctcgcac 
               
               
                   
               
               
                 caacaccaaccggtccagtacatgagccagtcgggtcactgaccccggcaccagtccctcagccactg 
               
               
                   
               
               
                 gatccagcgcccgcagtgactcccgaggccagtcacctgttggaggatcccgatgaagagacgagcca 
               
               
                   
               
               
                 ggctgtcaaagcccttcgggagatggccgatactgtgattccccagaaggaagaggctgcaatctgtg 
               
               
                   
               
               
                 gccaaatggacctttcccatccgcccccaaggggccatctggatgagctgacaaccacacttgagtcc 
               
               
                   
               
               
                 atgaccgaggatctgaacctggactcacccctgaccccggaattgaacgagattctggataccttcct 
               
            
           
         
       
     
     
       
         
           
               
               
               
             
               
                   
               
               
                 Name 
                 gRNA target/sequence 
                 gRNA 
               
               
                   
               
             
            
               
                 gRNA1 
                 tagtcttagagtatccagtg 
                 uagucuuagaguauccagug 
               
               
                 promoter 
                 (SEQ ID NO: 43) 
                 (SEQ ID NO: 49) 
               
               
                 HBG1/2 
                   
                   
               
               
                   
               
               
                 gRNA2 
                 ggctagggatgaagaataaa 
                 ggcuagggaugaagaauaaa 
               
               
                 promoter 
                 (SEQ ID NO: 44) 
                 (SEQ ID NO: 50) 
               
               
                 HBG1/2 
                   
                   
               
               
                   
               
               
                 gRNA1 HS2 
                 aatatgtcacattctgtctc 
                 aauaugucacauucugucuc 
               
               
                 enhancer 
                 (SEQ ID NO: 45) 
                 {SEQ ID NO: 51) 
               
               
                 HBG1/2 
                   
                   
               
               
                   
               
               
                 gRNA2 HS2 
                 ggactatgggaggtcactaa 
                 ggacuaugggaggucacuaa 
               
               
                 enhancer 
                 (SEQ ID NO: 46) 
                 (SEQ ID NO: 52) 
               
               
                 HBG1/2 
                   
                   
               
               
                   
               
               
                 gRNA3HS2 
                 gaaggttacacagaaccaga 
                 gaagguuacacagaaccaga 
               
               
                 enhancer 
                 (SEQ ID NO: 47) 
                 (SEQ ID NO: 53) 
               
               
                 HBG1/2 
                   
                   
               
               
                   
               
               
                 gRNA4 HS2 
                 gccctgtaagcatcctgctg 
                 gcCcuguaagcauccugcug 
               
               
                 enhancer 
                 (SEQ ID NO: 48) 
                 (SEQ ID NO: 54) 
               
               
                 HBG1/2 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
               
            
               
                 GS linker 
                   
               
               
                 SEQ ID NO: 55 
                   
               
               
                 (Gly-Gly-Gly-Gly-Ser) n , 
                   
               
               
                 wherein n is an integer between 0 and 10 
               
               
                   
               
               
                 Linker 
               
               
                 SEQ ID NO: 56 
                   
               
               
                 Gly-Gly-Gly-Gly-Gly 
                   
               
               
                   
               
               
                 Linker 
               
               
                 SEQ ID NO: 57 
                   
               
               
                 G1y-GIy-Ala-Gly Gly 
                   
               
               
                   
               
               
                 Linker 
               
               
                 SEQ ID NO: 58 
                   
               
               
                 Gly-G1y-Gly-Gly-Ser-Ser-Ser 
                   
               
               
                   
               
               
                 Linker 
               
               
                 SEQ ID NO: 59 
                   
               
               
                 Gly-Gly-Gly-Gly-Ala-Ala-Ala 
                   
               
               
                   
               
               
                 SV40 NLS 
               
               
                 SEQ ID NO: 60 
                   
               
               
                 Pro-Lys-Lys-Lys-Arg-Lys-Val 
                   
               
               
                   
               
               
                 DNA sequence of the gRNA constant region 
               
               
                 SEQ ID NO: 61 
                   
               
               
                 gtttaagagctatgctggaaacagcatagcaagtttaaataaggctagtccgttatcaactt 
                   
               
               
                   
               
               
                 gaaaaagtggcaccgagtcggtgc 
               
               
                   
               
               
                 RNA sequence of the gRNA constant region 
               
               
                 SEQ ID NO: 62 
                   
               
               
                 guuuaagagcuaugcuggaaacagcauagcaaguuuaaaaaaggcuaguccgauaacaacaa 
                   
               
               
                   
               
               
                 gaaaaagaggcaccgagacggagc 
               
               
                   
               
               
                 DNA sequence for VPH-dCas9-SS18 (In backbone pNi36): pNi95; lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9; capital no underline = SS18. 
               
               
                 SEQ ID NO: 63 
                   
               
               
                 
                   atggatgctttagacgattttgacttagatatgcttggttcagacgcgttagacgacttcgacctaga 
                 
                   
               
               
                   
               
               
                 
                   catgttaggctcagatgcattggacgacttcgatttagatatgttgggctccgatgccctagatgact 
                 
               
               
                   
               
               
                 
                   ttgatctagatatgctagggtcactacccagcgctagcgtcgagttcgaaggcagcggcgggccttca 
                 
               
               
                   
               
               
                 
                   gagcagatcagcaaccaggccctagctctggcccctagctcogctccagtgctggcccagactatggt 
                 
               
               
                   
               
               
                 
                   gccctctagtgctatggtgcctctggcccagccacctgctccagcccctgtgctgaccccaggaccac 
                 
               
               
                   
               
               
                 
                   cccagtcactgagcgctccagtgcccaagtctacacaggccggcgaggggactctgagtgaagctctg 
                 
               
               
                   
               
               
                 
                   ctgcacctgcagttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccggagt 
                 
               
               
                   
               
               
                 
                   gttcacagatctggcctccgtggacaactctgagtttcagcagctgctgaatcagggcgtgtccatgt 
                 
               
               
                   
               
               
                 
                   ctcatagtacagccgaaccaatgctgatggagtaccccgaagccattacccggctggtgaccggcagc 
                 
               
               
                   
               
               
                 
                   cagcggccccccgaccccgctccaactcccctgggaaccagcggcctgcctaatgggctgtccggaga 
                 
               
               
                   
               
               
                 
                   tgaagacttctcaagcatcgctgatatggactttagtgccctgctgtcacagatttcctctagtgggc 
                 
               
               
                   
               
               
                 
                   agggaggaggtggaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcccctcggtg 
                 
               
               
                   
               
               
                 
                   accgtgcccgacatgagcctgcctgaccttgacagcagcctggccagtatccaagagctcctatctcc 
                 
               
               
                   
               
               
                 
                   ccaggagccccccaggcctcccgaggcagagaacagcagcccggattcagggaagcagctggtgcact 
                 
               
               
                   
               
               
                 
                   acacagcgcagccgctgttcctgctggaccccggctccgtggacaccgggagcaacgacctgccggtg 
                 
               
               
                   
               
               
                 
                   ctgtttgagctgggagagggctcctacttctccgaaggggacggcttcgccgaggaccccaccatctc 
                 
               
               
                   
               
               
                   cctgctgacaggctcggagcctcccaaagccaaggaccccactgtctcc ggctctggaggatctggcg 
               
               
                   
               
               
                 gctctagcgccacc ATGGACAAGAAGTACAGCATCGGCCTGGCCATCGGCACCAACTCTGTGGGCTGG   
               
               
                   
               
               
                 
                   GCCGTGATCACCGACGAGTACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCA 
                 
               
               
                   
               
               
                 
                   CAGCATCAAGAAGAAGAACCTCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGC 
                 
               
               
                   
               
               
                 
                   TGAAGAGAACCGCCAGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTC 
                 
               
               
                   
               
               
                 
                   AGCAACGAGATGGCCAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGA 
                 
               
               
                   
               
               
                 
                   GGATAAGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGT 
                 
               
               
                   
               
               
                 
                   ACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATC 
                 
               
               
                   
               
               
                 
                   TATCTGGCCCTGGCCCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGA 
                 
               
               
                   
               
               
                 
                   CAACAGCGACGTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACC 
                 
               
               
                   
               
               
                 
                   CCATCAACGCCAGCGGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTG 
                 
               
               
                   
               
               
                 
                   GAAAATCTGATCGCCCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAG 
                 
               
               
                   
               
               
                 
                   CCTGGGCCTGACCCCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCA 
                 
               
               
                   
               
               
                 
                   AGGACACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTT 
                 
               
               
                   
               
               
                 
                   CTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCAC 
                 
               
               
                   
               
               
                 
                   CAAGGCCCCCCTGAGCGCCTCTATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGA 
                 
               
               
                   
               
               
                 
                   AAGCTCTCGTGCGGCAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGC 
                 
               
               
                   
               
               
                 
                   TACGCCGGCTACATTGACGGCGGAGCCAGCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGA 
                 
               
               
                   
               
               
                 
                   AAAGATGGACGGCACCGAGGAACTGCTCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGA 
                 
               
               
                   
               
               
                 
                   CCTTCGACAACGGLAGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAG 
                 
               
               
                   
               
               
                 
                   GAAGATTTTTACCCATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCC 
                 
               
               
                   
               
               
                 CTACTACGTGGGCCCTCTGGCCAGGGGAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAA 
               
               
                   
               
               
                 
                   CCATCACCCCCTGGAACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGG 
                 
               
               
                   
               
               
                 
                   ATGACCAACTTCGATAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTA 
                 
               
               
                   
               
               
                 
                   CTTCACCGTGTATAACGAGCTGACCAAAGTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCC 
                 
               
               
                   
               
               
                 
                   TGAGCGGCGAGCAGAAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAG 
                 
               
               
                   
               
               
                 
                   CAGCTGAAAGAGGACTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCCGuCGTGGAAGA 
                 
               
               
                   
               
               
                 
                   TCGGTTCAACGCCTCCCTGGGCACATACCACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGG 
                 
               
               
                   
               
               
                 
                   ACAATGAGGAAAACGAGGACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAG 
                 
               
               
                   
               
               
                 
                   ATGATCGAGGAACGGCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCG 
                 
               
               
                   
               
               
                 
                   GCGGAGATACACCGGCTGGGGCAGGCTGAGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCG 
                 
               
               
                   
               
               
                 
                   GCAAGACAATCCTGGATTTCCTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCAC 
                 
               
               
                   
               
               
                 
                   GACGACAGCCTGACCTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCA 
                 
               
               
                   
               
               
                 
                   CGAGCACATTGCCAATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGG 
                 
               
               
                   
               
               
                 
                   TGGACGAGCTCGTGAAAGTGATGGGCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAG 
                 
               
               
                   
               
               
                 
                   AACCAGACCACCCAGAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAA 
                 
               
               
                   
               
               
                 
                   AGAGCTGGGCAGCCAGATCCTGAAAGAACACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGT 
                 
               
               
                   
               
               
                 
                   ACCTGTACTACCTGCAGAATGGGCGGGATATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCC 
                 
               
               
                   
               
               
                 
                   GACTACGATGTGGACGCCATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCT 
                 
               
               
                   
               
               
                 
                   GACCAGAAGCGACAAGAACCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGA 
                 
               
               
                   
               
               
                 
                   AGAACTACTGGCGGCAGCTGCTGAACGCCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAG 
                 
               
               
                   
               
               
                 
                   GCCGAGAGAGGCGGCCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCG 
                 
               
               
                   
               
               
                 
                   GCAGATCACAAAGCACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACA 
                 
               
               
                   
               
               
                 
                   AGCTGATCCGGGAAGTGAAAGTGATCACCCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTC 
                 
               
               
                   
               
               
                 
                   CAGTTTTACAAAGTGCGCGAGATCAACAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGT 
                 
               
               
                   
               
               
                 
                   GGGAACCGCCCTGATCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGT 
                 
               
               
                   
               
               
                 
                   ACGACGTGCGGAAGATGATCGCCAAGAGCGAGCAGGAAATCGGCAAGGCTACCGCCAAGTACTTCTTC 
                 
               
               
                   
               
               
                 
                   TACAGCAACATCATGAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCC 
                 
               
               
                   
               
               
                 
                   TCTGATCGAGACAAACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGC 
                 
               
               
                   
               
               
                 
                   GGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGC 
                 
               
               
                   
               
               
                 
                   AAAGAGTCTATCCTGCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAA 
                 
               
               
                   
               
               
                 
                   GAAGTACGGCGGCTTCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGG 
                 
               
               
                   
               
               
                 
                   GCAAGTCCAAGAAACTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTC 
                 
               
               
                   
               
               
                 
                   GAGAAGAATCCCATCGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAA 
                 
               
               
                   
               
               
                 
                   GCTGCCTAAGTACTCCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAAC 
                 
               
               
                   
               
               
                 
                   TGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTAT 
                 
               
               
                   
               
               
                 
                   GAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTA 
                 
               
               
                   
               
               
                 
                   CCTGGACGAGATCATCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGG 
                 
               
               
                   
               
               
                 
                   ACAAAGTGCTGTCCGCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATC 
                 
               
               
                   
               
               
                 
                   CACCTGTTTACCCTGACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCG 
                 
               
               
                   
               
               
                 
                   GAAGAGGTACACCAGCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGT 
                 
               
               
                   
               
               
                   ACGAGACACGGATCGACCTGTCTCAGCTGGGAGGCGAC aagcgacctgccgccacaaagaaggctgga 
               
               
                   
               
               
                 caggctaagaagaagaaactggactctggaggatccgactacaaagaccatgacggtgattataaaga 
               
               
                   
               
               
                 tcatgacatcgattacaaggatgacgatgacaagggaggatccaaggagaagagtgcttgtcctaaag 
               
               
                   
               
               
                 atccagccaaacctccggccaaggcacaagttgtgggatggccaccggtgagatcataccggaagaac 
               
               
                   
               
               
                 gtgatggtttcctgccaaaaaccaagcggtggcccggaggcggcggcgctcgtgaaggtatcaatgga 
               
               
                   
               
               
                 cggagcaccgtacttgaggaaaatcgatttgaggatgtataaaggcggarctggcggctctggaggat 
               
               
                   
               
               
                 ccagcATGTCTGTGGCTTTCGCGGCCCCGAGGCAGCGAGGCAAGGGGGAGATCACTCCCGCTGCGATT 
               
               
                   
               
               
                 CAGAAGATGTTGGATGACAATAACCATCTTATTCAGTGTATAATGGACTCTCAGAATAAAGGAAAGAC 
               
               
                   
               
               
                 CTCAGAGTGTTCTCAGTATCAGCAGATGTTGCACACAAACTTGGTATACCTTGCTACAATAGCAGATT 
               
               
                   
               
               
                 CTAATCAAAATATGCAGTCTCTTTTACCAGCACCACCCACACAGAATATGCCTATGGGTCCTGGAGGG 
               
               
                   
               
               
                 ATGAATCAGAGCGGCCCTCCCCCACCTCCACGCTCTCACAACATGCCTTCAGATGGAATGGTAGGTGG 
               
               
                   
               
               
                 GGGTCCTCCTGCACCGCACATGCAGAACCAGATGAACGGCCAGATGCCTGGGCCTAACCATATGCCTA 
               
               
                   
               
               
                 TGCAGGGACCTGGACCCAATCAACTCAATATGACAAACAGTTCCATGAATATGCCTTCAAGTAGCCAT 
               
               
                   
               
               
                 GGATCCATGGGAGGTTACTACCATTCTGTGCCATCATCACAGAGCATGCCAGTACAGAATCAGATGAC 
               
               
                   
               
               
                 AATGAGTCAGGGACAACCAATGGGAAACTATGGTCCCAGACCAAATATGAGTATGCAGCCAAACCAAG 
               
               
                   
               
               
                 GTCCAATGATGCATCAGCAGCCTCCTTCTCAGCAATACAATATGCCACAGGGAGGCGGACAGCATTAC 
               
               
                   
               
               
                 CAAGGACAGCAGCCACCTATGGGAATGATGGGTCAAGTTAACCAAGGCAATCATATGATGGGTCAGAG 
               
               
                   
               
               
                 ACAGATTCCTCCCTATAGACCTCCTCAACAGGGCCCACCACAGCAGTACTCAGGCCAGGAAGACTATT 
               
               
                   
               
               
                 ACGGGGACCAATACAGTCATGGTGGACAAGGTCCTCCAGAACGCATGAACCAGCAATATTACCCTGAT 
               
               
                   
               
               
                 GGAAATTCACAGTATGGCCAACAGCAACATGCATACCAGGCACCACCTCCACAACAGGGATATCCACC 
               
               
                   
               
               
                 CCAGCACCAGCAGTACCCAGCGCAGCAAGGTTACCCAGGACAGCAGCAGGGCTACCGTCCTTCACAGG 
               
               
                   
               
               
                 GTGGTCCAGGTCCTCAGTATCCTAACTACCCACAGGGACAAGGTCAGCAGTATGGAGGATATAGACCA 
               
               
                   
               
               
                 ACACAGCCTGGACCACCACAGCCACCCCAGCAGAGGCCTTATGGATATGACCAGGGACAGTATGGAAA 
               
               
                   
               
               
                 TTACCAGCAGTGA 
               
               
                   
               
               
                 Amino acid sequence for VPH-dCas9-SS18 
               
               
                 (corresponding to SEQ ID NO: 63); lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9; 
               
               
                 capital no underline = SS18. 
               
               
                 SEQ ID NO: 64 
                   
               
               
                 
                   dalddfdldmlgsdalddfdldmlgsdalddfdldmlgsdalddfdldmlqslpsasvefegsgqpsg  
                 
                   
               
               
                   
               
               
                 
                   qisngalalapssapvlagtmvpssamvplagppapapvltpgppqslsapvpkstaagegllseall 
                 
               
               
                   
               
               
                 
                   hlqfdadedlgallgnstdpgvftdlasvdnsefqqllnqgvsmshstaepmlmeypeaitrlvtgsq 
                 
               
               
                   
               
               
                 
                   rppdpaptplgtsglpnglsgdedfssiadmdfsailsqisssgqqgggsgfsvdtsalldlfspsvt 
                 
               
               
                   
               
               
                 
                   vpdmslpdldsslasiqellspqepprppeaensspdsgkqlvhytaqplflldpgsvdtgsndlpvl 
                 
               
               
                   
               
               
                   felqegsyfseqdgfaedptislllqseppkakdptvs qsggsggssat MDKKYSIGLAIGTNSVGWA   
               
               
                   
               
               
                 
                   VITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFS 
                 
               
               
                   
               
               
                 
                   NEMAKVDDSFFHRLEESFLVEEDKKHEKHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIY 
                 
               
               
                   
               
               
                 
                   LALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE 
                 
               
               
                   
               
               
                 
                   NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFL 
                 
               
               
                   
               
               
                 
                   AAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQRLTLLKALVRQQLPEKYKEIFFDQSKNGY 
                 
               
               
                   
               
               
                 
                   AGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQE 
                 
               
               
                   
               
               
                 
                   DFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERM 
                 
               
               
                   
               
               
                 
                   TNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQ 
                 
               
               
                   
               
               
                 
                   LKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREM 
                 
               
               
                   
               
               
                 
                   IEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD 
                 
               
               
                   
               
               
                 
                   DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMAREN 
                 
               
               
                   
               
               
                 
                   QTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSD 
                 
               
               
                   
               
               
                 
                   YDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKA 
                 
               
               
                   
               
               
                 
                   ERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQ 
                 
               
               
                   
               
               
                 
                   FYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY 
                 
               
               
                   
               
               
                 
                   SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK 
                 
               
               
                   
               
               
                 
                   ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFE 
                 
               
               
                   
               
               
                 
                   KNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYE 
                 
               
               
                   
               
               
                 
                   KLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIH 
                 
               
               
                   
               
               
                   LFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD krpaatkkagq 
               
               
                   
               
               
                 akkkkldsggsdykdhdgdykdhdidykddddkggskeksacpkdpakppakaqvvgwppvrsyrknv 
               
               
                   
               
               
                 mvscqkssggpeaaafvkvsmdgapylrkidlrmykggsggsggssMSVAFAAPRQRGKGEITPAAIQ 
               
               
                   
               
               
                 KMLDDNNHLIQCIMDSQNKGKTSECSQYQQMLHTNLVYLATIADSNQNMQSLLPAPPTQNMPMGPGGM 
               
               
                   
               
               
                 NQSGPPPPPRSHNMPSDGMVGGGPPAPHMQNQMNGQMPGPNHMPMQGPGPNQLNMTNSSMNMPSSSHG 
               
               
                   
               
               
                 SMGGYNHSVPSSQSMPVQNQMTMSQGQPMGNYGPRPNMSMQPNQGPMMHQQPPSQQYNMPQGGGQHYQ 
               
               
                   
               
               
                 GQQPPMGMMGQVNQGNHMMGQRQIPPYRPPQQGPPQQYSGQEDYYGDQYSHGGQGPPEGMNQQYYPDG 
               
               
                   
               
               
                 NSQYGQQQDAYQGPPPQQGYPPQQQQYPGQQGYPGQQQGYGPSQGGPGPQYPNYPQGQGQQYGGYRPT 
               
               
                   
               
               
                 QPGPPQPPQQRPYGYDQGQYGNYQQ* 
               
               
                   
               
               
                 DNA sequence for VPH-dCas9-S818 (in backbone pNI144): pNH65; lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9; 
               
               
                 capital no underline = SS18. 
               
               
                 SEQ ID NO: 65 
                   
               
               
                 atggactacaaagaccatgacggtgattataaagatcatgacatcgattacaaggatgacgatgacaa 
                   
               
               
                   
               
               
                 gcacgtt gatgctttaggcgattttgacttagatatgcttggttcagacgcgttagacgacttcgacc   
               
               
                   
               
               
                 
                   tagacatgttaggctcagatgcattggacgacttcgatttagatatgttgggctccgatgccctagat 
                 
               
               
                   
               
               
                 
                   gactttgatctagatatgctagggtcactacccagcgccagcgtcgagttcgaaggcagcggcgggcc 
                 
               
               
                   
               
               
                 
                   ttcagggcagatcagcaaccaggccctggctctggcccctagctccgccccagtgctggcccagacta 
                 
               
               
                   
               
               
                 
                   tggtaccctctagtgctatggtgcctctggcccagccacctgctccagcccctgtgctgaccccagga 
                 
               
               
                   
               
               
                 
                   ccaccccagtcactgagcgccccagtgcccaagtctacacaggccggcgacgggactctgagtgaagc 
                 
               
               
                   
               
               
                 
                   tctgctgcacctgcagttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccg 
                 
               
               
                   
               
               
                 
                   gagtgttcacagatctggcctccgtggacaactctgagtttcagcagctgctgaatcagggcgtgtcc 
                 
               
               
                   
               
               
                 
                   atgtctcatagtacagccgaaccaatgctgatggagtaccccgaagccattacccggctggtgaccgg 
                 
               
               
                   
               
               
                 
                   cagccagcggccccccgaccccgctccaactcccctgggaaccagcggcctgcctaatgggctgtccg 
                 
               
               
                   
               
               
                 
                   gagatgaagacttctcaagcatcgctgatatggactttagtgccctgctgtcacagatttoctctagt 
                 
               
               
                   
               
               
                 
                   gggcagggaggaggtggaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcccctc 
                 
               
               
                   
               
               
                 
                   ggtgaccgtgcccgacatgagcctgcctgaccttgacagcagcctggccagtatccaagagctcctgt 
                 
               
               
                   
               
               
                 
                   ctccccaggagccccccaggcctcccgaggcagagaacagcagcccggattcagggaagcagctggtg 
                 
               
               
                   
               
               
                 
                   cactacacagcgcagccgctgttcctgctggaccccggctccgtggacaccgggagcaacgacctgcc 
                 
               
               
                   
               
               
                 
                   ggtgctgtttgagctgggagagggctcctacttctccgaaggggacggcttcgccgaggaccccacca 
                 
               
               
                   
               
               
                   tctccctgctgacaggctcggagcctcccaaagccaaggaccccactgtctcc aaccccaagaagaag 
               
               
                   
               
               
                 aggaaggtgggccgcgga ATGGACAAGAAGTAGTCCATTGGGCTCGCCATCGGCACAAACAGCGTCGG   
               
               
                   
               
               
                 
                   CTGGGCCGTCATTACGGACGAGTACAAGGTGCCGAGCAAAAAATTCAAAGTTCTGGGCAATACCGATC 
                 
               
               
                   
               
               
                 
                   GCCACAGCATAAAGAAGAACCTCATTGGCGCCCTCCTGTTCGACTCCGGGGAAACCGCCGAAGCCACG 
                 
               
               
                   
               
               
                 
                   CGGCTCAAAAGAACAGCACGGCGCAGATATACCCGCAGAAAGAATCGGATCTGCTACCTGCAGGAGAT 
                 
               
               
                   
               
               
                 
                   CTTTAGTAATGAGATGGCTAAGGTGGATGACTCTTTCTTCCATAGGCTGGAGGAGTCCTTTTTGGTGG 
                 
               
               
                   
               
               
                 
                   AGGAGGATAAAAAGCACGAGCGCCACCCAATCTTTGGCAATATCGTGGACGAGGTGGCGTACCATGAA 
                 
               
               
                   
               
               
                 
                   AAGTACCCAACCATATATCATCTGAGGAAGAAGCTTGTAGACAGTACTGATAAGGCTGACTTGCGGTT 
                 
               
               
                   
               
               
                 
                   GATCTATCTcGCGCTGGCGCATATGATCAAATTTCGGGGACACTTCCTCATCGAGGGGGACCTGAACC 
                 
               
               
                   
               
               
                 
                   CAGACAACAGCGATGTCGACAAACTCTTTATCCAACTGGTTCAGACTTACAATCAGCTTTTCGAAGAG 
                 
               
               
                   
               
               
                 
                   AACCCGATCAACGCATCCGGAGTTGACGCCAAAGCAATCCTGAGCGCTAGGCTGTCCAAATCCCGGCG 
                 
               
               
                   
               
               
                 
                   GCTCGAAAACCTCATCGCACAGCTCCCTGGGGAGAAGAAGAACGGCCTGTTTGGTAATCTTATCGCCC 
                 
               
               
                   
               
               
                 
                   TGTCACTCGGGCTGACCCCCAACTTTAAATCTAACTTCGACCTGGCCGAAGATGCCAAGCTTCAACTG 
                 
               
               
                   
               
               
                 
                   AGCAAAGACACCTACGATGATGATCTCGACAATCTGCTGGCCCAGATCGGCGACCAGTACGCAGACCT 
                 
               
               
                   
               
               
                 
                   TTTTTTGGCGGCAAAGAACCTGTCAGACGCCATTCTGCTGAGTGATATTCTGCGAGTGAACACGGAGA 
                 
               
               
                   
               
               
                 
                   TCACCAAAGCTCCGCTGAGCGCTAGTATGATCAAGCGCTATGATGAGCACCACCAAGACTTGACTTTG 
                 
               
               
                   
               
               
                 
                   CTGAAGGCCCTTGTCAGACAGCAACTGCCTGAGAAGTACAAGGAAATTTTCTTCGATCAGTCTAAAAA 
                 
               
               
                   
               
               
                 
                   TGGCTACGCCGGATACATTGACGGCGGAGCAAGCCAGGAGGAATTTTACAAATTTATTAAGCCCATCT 
                 
               
               
                   
               
               
                 
                   TGGAAAAAATGGACGGCACCGAGGAGCTGCTGGTAAAGCTTAACAGAGAAGATCTGTTGCGCAAACAG 
                 
               
               
                   
               
               
                 
                   CGCACTTTCGACAATGGAAGCATCCCCCACCAGATTCACCTGGGCGAACTGCACGCTATCCTCAGGCG 
                 
               
               
                   
               
               
                 
                   GCAAGAGGATTTCTACCCCTTTTTGAAAGATAACAGGGAAAAGATTGAGAAAATCCTCACATTTCGGA 
                 
               
               
                   
               
               
                 
                   TACCCTACTATGTAGGCCCCCTCGCCCGGGGAAATTCCAGATTCGCGTGGATGACTCGCAAATCAGAA 
                 
               
               
                   
               
               
                 
                   GAGACCATCACTCCCTGGAACTTCGAGGAAGTCGTGGATAAGGGGGCCTCTGCCCAGTCCTTCATCGA 
                 
               
               
                   
               
               
                 
                   AAGGATGACTAACTTTGATAAAAATCTGCCTAACGAAAAGGTGCTTCCTAAACACTCTCTGCTGTACG 
                 
               
               
                   
               
               
                 
                   AGTACTTCACAGTTTATAACGAGCTCACCAAGGTCAAATACGTCACAGAAGGGATGAGAAAGCCAGCA 
                 
               
               
                   
               
               
                 
                   TTCCTGTCTGGAGAGCAGAAGAAAGCTATCGTGGACCTCCTCTTCAAGACGAACCGGAAAGTTACCGT 
                 
               
               
                   
               
               
                 
                   GAAACAGCTCAAAGAAGACTATTTCAAAAAGATTGAATGTTTCGACTCTGTTGAAATCAGCGGAGTGG 
                 
               
               
                   
               
               
                 
                   AGGATCGCTTCAACGCATCCCTGGGAACGTATCACGATCTCCTGAAAATCATTAAAGACAAGGACTTC 
                 
               
               
                   
               
               
                 
                   CTGGACAATGAGGAGAACGAGGACATTCTTGAGGACATTGTCCTCACCCTTACGTTGTTTGAAGATAG 
                 
               
               
                   
               
               
                 
                   GGAGATGATTGAAGAACGCTTGAAAACTTACGCTCATCTCTTCGACGACAAAGTCATGAAACAGCTCA 
                 
               
               
                   
               
               
                 
                   AGAGGCGCCGATATACAGGATGGGGGCGGCTGTCAAGAAAACTGATCAATGGGATCCGAGACAAGCAG 
                 
               
               
                   
               
               
                 
                   AGTGGAAAGACAATCCTGGATTTTCTTAAGTCCGATGGATTTGCCAACCGGAACTTCATGCAGTTGAT 
                 
               
               
                   
               
               
                 
                   CCATGATGACTCTCTCACCTTTAAGGAGGACATCCAGAAAGCACAAGTTTCTGGCCAGGGGGACAGTC 
                 
               
               
                   
               
               
                 
                   TTCACGAGCACATCGCTAATCTTGCAGGTAGCCCAGCTATCAAAAAGGGAATACTGCAGACCGTTAAG 
                 
               
               
                   
               
               
                 
                   GTCGTGGATGAACTCGTCAAAGTAATGGGAAGGCATAAGCCCGAGAATATCGTTATCGAGATGGCCCG 
                 
               
               
                   
               
               
                 
                   AGAGAACCAAACTACCCAGAAGGGACAGAAGAACAGTAGGGAAAGGATGAAGAGGATTGAAGAGGGTA 
                 
               
               
                   
               
               
                 
                   TAAAAGAACTGGGGTCCCAAATCCTTAAGGAACACCCAGTTGAAAACACCCAGCTTCAGAATGAGAAG 
                 
               
               
                   
               
               
                 
                   CTCTACCTGTACTACCTGCAGAACGGCAGGGACATGTACGTGGATCAGGAACTGGACATCAATCGGCT 
                 
               
               
                   
               
               
                 
                   CTCCGACTACGACGTGGATGCCATCGTGCCCCAGTCTTTTCTCAAAGATGATTCTATTGATAATAAAG 
                 
               
               
                   
               
               
                 
                   TGTTGACAAGATCCGATAAAAATAGAGGGAAGAGTGATAACGTCCCCTCAGAAGAAGTTGTCAAGAAA 
                 
               
               
                   
               
               
                 
                   ATGAAAAATTATTGGCGGCAGCTGCTGAACGCCAAACTGATCACACAACGGAAGTTCGATAATCTGAC 
                 
               
               
                   
               
               
                 
                   TAAGGCTGAACGAGGTGGCCTGTCTGAGTTGGATAAAGCCGGCTTCATCAAAAGGCAGCTTGTTGAGA 
                 
               
               
                   
               
               
                 
                   CACGCCAGATCACCAAGCACGTGGCCCAAATTCTCGATTCACGCATGAACACCAAGTACGATGAAAAT 
                 
               
               
                   
               
               
                 
                   GACAAACTGATTCGAGAGGTGAAAGTTATTACTCTGAAGTCTAAGCTGGTCTCAGATTTCAGAAAGGA 
                 
               
               
                   
               
               
                 
                   CTTTCAGTTTTATAAGGTGAGAGAGATCAACAATTACCACCATGCGCATGATGCCTACCTGAATGCAG 
                 
               
               
                   
               
               
                 
                   TGGTAGGCACTGCACTTATCAAAAAATATCCCAAGCTTGAATCTGAATTTGTTTACGGAGACTATAAA 
                 
               
               
                   
               
               
                 
                   GTGTACGATGTTAGGAAAATGATCGCAAAGTCTGAGCAGGAAATAGGCAAGGCCACCGCTAAGTACTT 
                 
               
               
                   
               
               
                 
                   CTTTTACAGCAATATTATGAATTTTTTCAAGACCGAGATTACACTGGCCAATGGAGAGATTCGGAAGC 
                 
               
               
                   
               
               
                 
                   GACCACTTATCGAAACAAACGGAGAAACAGGAGAAATCGTGTGGGACAAGGGTAGGGATTTCGCGACA 
                 
               
               
                   
               
               
                 
                   GTCCGGAAGGTCCTGTCCATGCCGCAGGTGAACATCGTTAAAAAGACCGAAGTACAGACCGGAGGCTT 
                 
               
               
                   
               
               
                 
                   CTCCAAGGAAAGTATCCTCCCGAAAAGGAACAGCGACAAGCTGATCGCACGCAAAAAAGATTGGGACC 
                 
               
               
                   
               
               
                 
                   CCAAGAAATACGGCGGATTCGATTCTCCTACAGTCGCTTACAGTGTACTGGTTGTGGCCAAAGTGGAG 
                 
               
               
                   
               
               
                 
                   AAAGGGAAGTCTAAAAAACTCAAAAGCGTCAAGGAACTGCTGGGCATCACAATCATGGAGCGATCAAG 
                 
               
               
                   
               
               
                 
                   CTTCGAAAAAAACCCCATCGACTTTCTCGAGGCGAAAGGATATAAAGAGGTCAAAAAAGACCTCATCA 
                 
               
               
                   
               
               
                 
                   TTAAGCTTCCCAAGTACTCTCTCTTTGAGCTTGAAAACGGCCGGAAACGAATGCTCGCTAGTGCGGGC 
                 
               
               
                   
               
               
                 
                   GAGCTGCAGAAAGGTAACGAGCTGGCACTGCCCTCTAAATACGTTAATTTCTTGTATCTGGCCAGCCA 
                 
               
               
                   
               
               
                 
                   CTATGAAAAGCTCAAAGGGTCTCCCGAAGATAATGAGCAGAAGCAGCTGTTCGTGGAACAACACAAAC 
                 
               
               
                   
               
               
                 
                   ACTACCTTGATGAGATCATCGAGCAAATAAGCGAATTCTCCAAAAGAGTGATCCTCGCCGACGCTAAC 
                 
               
               
                   
               
               
                 
                   CTCGATAAGGTGCTTTCTGCTTACAATAAGCACAGGGATAAGCCCATCAGGGAGCAGGCAGAAAACAT 
                 
               
               
                   
               
               
                 
                   TATCCACTTGTTTACTCTGACCAACTTGGGCGCGCCTGCAGCCTTCAAGTACTTCGACACCACCATAG 
                 
               
               
                   
               
               
                 
                   ACAGAAAGCGGTACACCTCTACAAAGGAGGTCCTGGACGCCACACTGATTCATCAGTCAATTACGGGG 
                 
               
               
                   
               
               
                   CTCTATGAAACAAGAATCGACCTCTCTCAGCTCGGTGGAGACAGCAGGGCTGAC cccaagaagaagag 
               
               
                   
               
               
                 gaaggtggctagcATGTCTGTGGCTTTCGCGGCCCCGAGGCAGCGAGGCAAGGGGGAGATCACTCCCG 
               
               
                   
               
               
                 CTGCGATTCAGAAGATGTTGGATGACAATAACCATCTTATTCAGTGTATAATGGACTCTCAGAATAAA 
               
               
                   
               
               
                 GGAAAGACCTCAGAGTGTTCTCAGTATCAGCAGATGTTGCACACAAACTTGGTATACCTTGCTACAAT 
               
               
                   
               
               
                 AGCAGATTCTAATCAAAATATGCAGTCTCTTTTACCAGCACCACCCACACAGAATATGCCTATGGGTC 
               
               
                   
               
               
                 CTGGAGGGATGAATCAGAGCGGCCCTCCCCCACCTCCACGCTCTCACAACATGCCTTCAGATGGAATG 
               
               
                   
               
               
                 GTAGGTGGGGGTCCTCCTGCACCGCACATGCAGAACCAGATGAACGGCCAGATGCCTGGGCCTAACCA 
               
               
                   
               
               
                 TATGCCTATGCAGGGACCTGGACCCAATCAACTCAATATGACAAACAGTTCCATGAATATGCCTTCAA 
               
               
                   
               
               
                 GTAGCCATGGATCCATGGGAGGTTACAACCATTCTGTGCCATCATCACAGAGCATGCCAGTACAGAAT 
               
               
                   
               
               
                 CAGATGACAATGAGTCAGGGACAACCAATGGGAAACTATGGTCCCAGACCAAATATGAGTATGCAGCC 
               
               
                   
               
               
                 AAACCAAGGTCCAATGATGCATCAGCAGCCTCCTTCTCAGCAATACAATATGCCACAGGGAGGCGGAC 
               
               
                   
               
               
                 AGCATTACCAAGGACAGCAGCCACCTATGGGAATGATGGGTCAAGTTAACCAAGGCAATCATATGATG 
               
               
                   
               
               
                 GGTCAGAGACAGATTCCTCCCTATAGACCTCCTCAACAGGGCCCACCACAGCAGTACTCAGGCCAGGA 
               
               
                   
               
               
                 AGACTATTACGGGGACCAATACAGTCATGGTGGACAAGGTCCTCCAGAAGGCATGAACCAGCAATATT 
               
               
                   
               
               
                 ACCCTGATGGAAATTCACAGTATGGCCAACAGCAAGATGCATACCAGGGACCACCTCCACAACAGGGA 
               
               
                   
               
               
                 TATCCACCCCAGCAGCAGCAGTACCCAGGGCAGCAAGGTTACCCAGGACAGCAGCAGGGCTACGGTCC 
               
               
                   
               
               
                 TTCACAGGGTGGTCCAGGTCCTCAGTATCCTAACTACCCACAGGGACAAGGTCAGCAGTATGGAGGAT 
               
               
                   
               
               
                 ATAGACCAACACAGCCTGGACCACCACAGCCACCCCAGCAGAGGCCTTATGGATATGACCAGGGACAG 
               
               
                   
               
               
                 TATGGAAATTACCAGCAGTGA 
               
               
                   
               
               
                 Amino acid sequence for VPH-dCas9-SS18 
               
               
                 (corresponding to SEQ ID NO: 65); lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9; 
               
               
                 capital no underline = SS18. 
               
               
                 SEQ ID NO: 66 
                   
               
               
                 mdykdhdqdykdhdidykddddkhv dalddfdldmlqsdalddfdldmlqsdalddfdldmlqsdald   
                   
               
               
                   
               
               
                 
                   dfdldmlqslpsasvefegsqqpsgqisnqalalapssapvlaqtmvpssamvplaqppapapvltpg 
                 
               
               
                   
               
               
                 
                   ppgslsapvpkstqagegtlseallhlqfdadedlgallqnstdpgvftdlasvdnsefqqllnqgvs 
                 
               
               
                   
               
               
                 
                   mshstaepmlmeypeaitrlvtqsqrppdpaptplqtsglpnglsgdedfssiadmdfsallsqisss 
                 
               
               
                   
               
               
                 
                   qqggggsgfsvdtsalldlfspsvtvpdmslpdldsslasiqellspqepprppeaensspdsgkqlv 
                 
               
               
                   
               
               
                   hytaqplflldpgsvdtgsndlpvlfelgegsyfsegdgfaedptislltgseppkakdptvs npkkk 
               
               
                   
               
               
                 rkvgrg MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT   
               
               
                   
               
               
                 
                   RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHE 
                 
               
               
                   
               
               
                 
                   KYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEE 
                 
               
               
                   
               
               
                 
                   NPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQL 
                 
               
               
                   
               
               
                 
                   SKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTL 
                 
               
               
                   
               
               
                 
                   LKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQ 
                 
               
               
                   
               
               
                 
                   RTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE 
                 
               
               
                   
               
               
                 
                   ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPA 
                 
               
               
                   
               
               
                 
                   FLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDF 
                 
               
               
                   
               
               
                 
                   LDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQ 
                 
               
               
                   
               
               
                 
                   SGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVK 
                 
               
               
                   
               
               
                 
                   VVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEK 
                 
               
               
                   
               
               
                 
                   LYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKK 
                 
               
               
                   
               
               
                 
                   MKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDEN 
                 
               
               
                   
               
               
                 
                   DKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYK 
                 
               
               
                   
               
               
                 
                   VYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFAT 
                 
               
               
                   
               
               
                 
                   VRKVLSMPQVNTVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVE 
                 
               
               
                   
               
               
                 
                   KGKSKKLKSVKELLGITIMERSSFEKNPIRFLEAKGYKEVKKDLlIKLPKYSLFELENGRKRMLASAG 
                 
               
               
                   
               
               
                 
                   ELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADAN 
                 
               
               
                   
               
               
                 
                   LDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITG 
                 
               
               
                   
               
               
                   LYETRIDLSQLGGDSRAD pkkkrkvasMSVAFAAPRQRGKGEITPAAIQKMLDDNNHLIQCIMDSQNK 
               
               
                   
               
               
                 GKTSECSQYQQMLHTNLVYLATIADSNQNMQSLLPAPPTONMPMGPGGMNQSGPPPPPRSHNMPSDGM 
               
               
                   
               
               
                 VGGGPPAPHMQNQMNGQMPGPNHMPMQGPGPNQLNMTNSSMNMPSSSHGSMGGYNHSVPSSQSMPVQN 
               
               
                   
               
               
                 QMTMSQGQPMGNYGPRPNMSMQPNQGPMMHQQPPSQQYNMPQGGGQHYQGQQPPMGMMGQVNQGNHMM 
               
               
                   
               
               
                 GQRQIPPYRPPQQGPPQQYSGQEDYYGDQYSHGGQGPPEGMNQQYYPDGNSQYGQQQDAYQGPPPQQG 
               
               
                   
               
               
                 YPPQQQQYPGQQGYPGQQQGYGPSQGGPGPQYPNYPQGQGQQYGGYRPTQPGPPQPPQQRPYGYDQGQ 
               
               
                   
               
               
                 YGNYQQ 
               
               
                   
               
               
                 DNA sequence for VPH-dCas9 (in backbone pNI36): 
               
               
                 pNI114; lowercase underlined = VPH; 
               
               
                 capital underlined = dCas9. 
               
               
                 SEQ ID NO: 67 
                   
               
               
                 
                   atggatgctttagacgattttgacttagatatgcttggttcagacgcgttagacgacttcgacctaga 
                 
                   
               
               
                   
               
               
                 
                   catgttaggctcagatgcattggacgacttcgatttagatatgttgggctccgatgccctagatgact 
                 
               
               
                   
               
               
                 
                   ttgatctagatatgctagggtcactacccagcgccagcgtcgagttcgaaggcagcggcgggccttca 
                 
               
               
                   
               
               
                 
                   gggcagatcagcaaccaggccctggctctggcccctagctccgctccagtgctggcccagactatggt 
                 
               
               
                   
               
               
                 
                   gccctctagtgctatggtgcctctggcccagccacctgctccagcccctgtgctgaccccaggaccac 
                 
               
               
                   
               
               
                 
                   cccagtcactgagcgccccagtgcccaagtctacacaggccggcgaggggactctgagtgaagctctg 
                 
               
               
                   
               
               
                 
                   ctgcacctgcagttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccggagt 
                 
               
               
                   
               
               
                 
                   gttcacagatctggcctccgtggacaactctgagtttcagcagctgctgaatcagggcgtgtccatgt 
                 
               
               
                   
               
               
                 
                   ctcatagtacagccgaaccaatgctgatggagtaccccgaagccattacccggctggtgaccggcagc 
                 
               
               
                   
               
               
                 
                   cagcggccccccgaccccgctccaactcccctgggaaccagcggcctgcctaatgggctgtccggaga 
                 
               
               
                   
               
               
                 
                   tgaagacttctcaagcatogctgatatggactttagtgccctgctgtcacagatttcctctagtgggc 
                 
               
               
                   
               
               
                 
                   agggaggaggtggaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcccctcggtg 
                 
               
               
                   
               
               
                 
                   accgtgcccgacatgagcctgcctgaccttgacagcagcctggccagtatccaagagctcctgtctcc 
                 
               
               
                   
               
               
                 
                   ccaggagccccccaggcctcccgaggcagagaacagcagcccggattcagggaagcagctggtgcact 
                 
               
               
                   
               
               
                 
                   acacagcgcagccgctgttcctgctggaccccggctccgtggacaccgggagcaacgacctgccggtg 
                 
               
               
                   
               
               
                 
                   ctgtttgagctgggagagggctcctacttctccgaaggggacggcttcgccgaggaccccaccatctc 
                 
               
               
                   
               
               
                   cctgctgacaggctcggagcctcccaaagccaaggaccccactgtctcc ggctctggaggatctggcg 
               
               
                   
               
               
                 gctctagcgccacc ATGGACAAGAAGTACAGCATCGGCCTGGCCATCGGCACCAACTCTGTGGGCTGG   
               
               
                   
               
               
                 
                   GCCGTGATCACCGACGAGTACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCA 
                 
               
               
                   
               
               
                 
                   CAGCATCAAGAAGAACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGC 
                 
               
               
                   
               
               
                 
                   TGAAGAGAACCGCCAGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTC 
                 
               
               
                   
               
               
                 AGCAACGAGATGGCCAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGA 
               
               
                   
               
               
                 
                   GGATAAGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGT 
                 
               
               
                   
               
               
                 
                   ACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATC 
                 
               
               
                   
               
               
                 
                   TATCTGGCCCTGGCCCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGA 
                 
               
               
                   
               
               
                 
                   CAACAGCGACGTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACC 
                 
               
               
                   
               
               
                 
                   CCATCAACGCCAGCGGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTG 
                 
               
               
                   
               
               
                 
                   GAAAATCTGATCGCCCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAG 
                 
               
               
                   
               
               
                 
                   CCTGGGCCTGACCCCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCA 
                 
               
               
                   
               
               
                 
                   AGGACACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTT 
                 
               
               
                   
               
               
                 
                   CTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCAC 
                 
               
               
                   
               
               
                 
                   CAAGGCCCCCCTGAGCGCCTCTATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGA 
                 
               
               
                   
               
               
                 
                   AAGCTCTCGTGCGGCAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGC 
                 
               
               
                   
               
               
                 
                   TACGCCGGCTACATTGACGGCGGAGCCAGCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGA 
                 
               
               
                   
               
               
                 
                   AAAGATGGACGGCACCGAGGAACTGCTCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGA 
                 
               
               
                   
               
               
                 
                   CCTTCGACAACGGCAGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAG 
                 
               
               
                   
               
               
                 
                   GAAGATTTTTACCCATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCC 
                 
               
               
                   
               
               
                 
                   CTACTACGTGGGCCCTCTGGCCAGGGGAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAA 
                 
               
               
                   
               
               
                 
                   CCATCACCCCCTGGAACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGG 
                 
               
               
                   
               
               
                 
                   ATGACCAACTTCGATAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTA 
                 
               
               
                   
               
               
                 
                   CTTCACCGTGTATAACGAGCTGACCAAAGTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCC 
                 
               
               
                   
               
               
                 
                   TGAGCGGCGAGCAGAAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAG 
                 
               
               
                   
               
               
                 
                   CAGCTGAAAGAGGACTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCCGGCGTGGAAGA 
                 
               
               
                   
               
               
                 
                   TCGGTTCAACGCCTCCCTGGGCACATACCACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGG 
                 
               
               
                   
               
               
                 
                   ACAATGAGGAAAACGAGGACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAG 
                 
               
               
                   
               
               
                 
                   ATGATCGAGGAACGGCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCG 
                 
               
               
                   
               
               
                 
                   GCGGAGATACACCGGCTGGGGCAGGCTGAGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCG 
                 
               
               
                   
               
               
                 
                   GCAAGACAATCCTGGATTTCCTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCAC 
                 
               
               
                   
               
               
                 
                   GACGACAGCCTGACCTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCA 
                 
               
               
                   
               
               
                 
                   CGAGCACATTGCCAATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGG 
                 
               
               
                   
               
               
                 
                   TGGACGAGCTCGTGAAAGTGATGGGCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAG 
                 
               
               
                   
               
               
                 
                   AACCAGACCACCCAGAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAA 
                 
               
               
                   
               
               
                 
                   AGAGCTGGGCAGCCAGATCCTGAAAGAACACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGT 
                 
               
               
                   
               
               
                 
                   ACCTGTACTACCTGCAGAATGGGCGGGATATGTACGTGGACGAGGAACTGGACATCAACCGGCTGTCC 
                 
               
               
                   
               
               
                 
                   GACTACGATGTGGACGCCATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCT 
                 
               
               
                   
               
               
                 
                   GACCAGAAGCGACAAGAACCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGA 
                 
               
               
                   
               
               
                 
                   AGAACTACTGGCGGCAGCTGCTGAACGCCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAG 
                 
               
               
                   
               
               
                 
                   GCCGAGAGAGGCGGCCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCG 
                 
               
               
                   
               
               
                 
                   GCAGATCACAAAGCACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACA 
                 
               
               
                   
               
               
                 
                   AGCTGATCCGGGAAGTGAAAGTGATCACCCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTC 
                 
               
               
                   
               
               
                 
                   CAGTTTTACAAAGTGCGCGAGATCAACAACTACCACCACGCLCACGACGCCTACCTGAACGCCGTCGt 
                 
               
               
                   
               
               
                 
                   GGGAACCGCCCTGATCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGT 
                 
               
               
                   
               
               
                 
                   ACGACGTGCGGAAGATGATCGCCAAGAGCGAGCAGGAAATCGGCAAGGCTACCGCCAAGTACTTCTTC 
                 
               
               
                   
               
               
                 
                   TACAGCAACATCATGAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCC 
                 
               
               
                   
               
               
                 
                   TCTGATCGAGACAAACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGC 
                 
               
               
                   
               
               
                 
                   GGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGC 
                 
               
               
                   
               
               
                 
                   AAAGAGTCTATCCTGCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAA 
                 
               
               
                   
               
               
                 
                   GAAGTACGGCGGCTTCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGG 
                 
               
               
                   
               
               
                 
                   GCAAGTCCAAGAAACTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTC 
                 
               
               
                   
               
               
                 
                   GAGAAGAATCCCATCGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAA 
                 
               
               
                   
               
               
                 
                   GCTGCCTAAGTACTCCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAAC 
                 
               
               
                   
               
               
                 
                   TGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTAT 
                 
               
               
                   
               
               
                 
                   GAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTA 
                 
               
               
                   
               
               
                 
                   CCTGGACGAGATCATCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGG 
                 
               
               
                   
               
               
                 
                   ACAAAGTGCTGTCCGCCTAGAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATC 
                 
               
               
                   
               
               
                 
                   CACCTGTTTACCCTGACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCG 
                 
               
               
                   
               
               
                 
                   GAAGAGGTACACCAGCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGT 
                 
               
               
                   
               
               
                 
                   ACGAGACACGGATCGACCTGTCTCAGCTGGGAGGCGAC 
                 
               
               
                   
               
               
                 Amino acid sequence for VPH-dCas9 
               
               
                 (corresponding to SEQ ID NO: 67); lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 68 
                   
               
               
                 
                   dalddfdldmlgsdalddfdldmlgsdalddfdldmlgsdalddfdldmlgslpsasvefegsggpsg 
                 
                   
               
               
                   
               
               
                 
                   qisnqalalapssapvlaqtmvpssamvplaqppapapvltpgppqslsapvpkstqaqegllseall 
                 
               
               
                   
               
               
                 
                   hlqfdadedlgallgnstdpgvftdlasvdnsefgqlinqgvsmshstaepmlmeypeaitrlvtgsq 
                 
               
               
                   
               
               
                 
                   rppdpaptplgtsglpnglsgdedfssiadmdfsallsqisssgqggggsgfsvdtsalldlfspsvt 
                 
               
               
                   
               
               
                 
                   vpdmslpdldsslasiqellspqepprppeaensspdsgkqlvhytaqplflldpgsvdtgsndlpvl 
                 
               
               
                   
               
               
                   felgeqsyfsegdgfaedptislltgseppkakdptvs gsqgsggssat MDKKYSIGLAIGTNSVGWA   
               
               
                   
               
               
                 
                   VITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFS 
                 
               
               
                   
               
               
                 
                   NEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIY 
                 
               
               
                   
               
               
                 
                   LALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE 
                 
               
               
                   
               
               
                 
                   NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFL 
                 
               
               
                   
               
               
                 
                   AAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY 
                 
               
               
                   
               
               
                 
                   AGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQE 
                 
               
               
                   
               
               
                 
                   DFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERM 
                 
               
               
                   
               
               
                 
                   TNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQ 
                 
               
               
                   
               
               
                 
                   LKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREM 
                 
               
               
                   
               
               
                 
                   IEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD 
                 
               
               
                   
               
               
                 
                   DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMAREN 
                 
               
               
                   
               
               
                 
                   QTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSD 
                 
               
               
                   
               
               
                 
                   YDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKA 
                 
               
               
                   
               
               
                 
                   ERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQ 
                 
               
               
                   
               
               
                 
                   FYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY 
                 
               
               
                   
               
               
                 
                   SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK 
                 
               
               
                   
               
               
                 
                   ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFE 
                 
               
               
                   
               
               
                 
                   KNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYE 
                 
               
               
                   
               
               
                 
                   KLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIH 
                 
               
               
                   
               
               
                 
                   LFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD 
                 
               
               
                   
               
               
                 DNA sequence for VPH-dCas9 (in backbone pNI123): 
               
               
                 pNI136; lowercase underlined = VPH; 
               
               
                 capital underlined = dCas9. 
               
               
                 SEQ ID NO: 69 
                   
               
               
                 atggactacaaagaccacgacggCgattataaagatcacgacatcgattacaaggatgacgatgacaa 
                   
               
               
                   
               
               
                 gcacgtt gatgctttagacgattttgacttagatatgcttggttcagacgcgttagacgacttcgacc   
               
               
                   
               
               
                 
                   tagacatgttaggctcagatgcattggacgacttcgatttagatatgttgggctccgatgccctagat 
                 
               
               
                   
               
               
                 
                   gactttgatctagatatgctagggtcactacccagcgccagcgtcgagttcgaaggcagcggcgggcc 
                 
               
               
                   
               
               
                 
                   ttcagggcagatcagcaaccaggccctggctctggcccctagctccgctccagtgctggcccagacta 
                 
               
               
                   
               
               
                 
                   tggtgccctctagtgctatggtgcctctggcccagccacctgctccagcccctgtgctgaccccagga 
                 
               
               
                   
               
               
                 
                   ccaccccagtcactgagcgccccagtgcccaagtctacacaggccggcgaggggactctgagtgaagc 
                 
               
               
                   
               
               
                 
                   tctgctgcacctgcagttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccg 
                 
               
               
                   
               
               
                 
                   gagtgttcacagatctggcctccgtggacaactctgagtttcagcagctgctgaatcagggcgtgtcc 
                 
               
               
                   
               
               
                 
                   atgtctcatagtacagccgaaccaatgctgatggagtaccccgaagccattacccggctggtgaccgg 
                 
               
               
                   
               
               
                 
                   cagccagcggccccccgaccccgctccaactcccctgggaaccagcggcctgcctaatgggctgtccg 
                 
               
               
                   
               
               
                 
                   gagatgaagacttctcaagcatcgctgatatggactttagtgccctgctgtcacagatttcctctagt 
                 
               
               
                   
               
               
                 
                   gggcagggaggaggtggaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcccctc 
                 
               
               
                   
               
               
                 
                   ggtgaccgtgcccgacatgagcctgcctgaccttgacagcagcctggccagtatccaagagctcctgt 
                 
               
               
                   
               
               
                 
                   ctccccaggagccccccaggcctcccgaggcagagaacagcagcccggattcagggaagcagctggtg 
                 
               
               
                   
               
               
                 
                   cactacacagcgcagccgctgttcctgctggaccccggctccgtggacaccgggagcaacgacctgcc 
                 
               
               
                   
               
               
                 
                   ggtgctgtttgagctgggagagggctcctacttctccgaaggggacggcttcgccgaggaccccacca 
                 
               
               
                   
               
               
                   tctccctgctgacaggctcggagcctcccaaagccaaggaccccactgtctcc aaccccaagaagaag 
               
               
                   
               
               
                 aggaaggtgggccgcgga ATGGACAAGAAGTACTCCATTGGGCTCGCCATCGGCACAAACAGCGTCGG   
               
               
                   
               
               
                 
                   CTGGGCCGTCATTACGGACGAGTACAAGGTGCCGAGCAAAAAATTCAAAGTTCTGGGCAATACCGATC 
                 
               
               
                   
               
               
                 
                   GCCACAGCATAAAGAAGAACCTCATTGGCGCCCTCCTGTTCGACTCCGGGGAAACCGCCGAAGCCACG 
                 
               
               
                   
               
               
                 
                   CGGCTCAAAAGAACAGCACGGCGCAGATATACCCGCAGAAAGAATCGGATCTGCTACCTGCAGGAGAT 
                 
               
               
                   
               
               
                 
                   CTTTAGTAATGAGATGGCTAAGGTGGATGACTCTTTCTTCCATAGGCTGGAGGAGTCCTTTTTGGTGG 
                 
               
               
                   
               
               
                 
                   AGGAGGATAAAAAGCACGAGCGCCACCCAATCTTTGGCAATATCGTGGACGAGGTGGCGTACCATGAA 
                 
               
               
                   
               
               
                 
                   AAGTACCCAACCATATATCATCTGAGGAAGAAGCTTGTAGACAGTACTGATAAGGCTGACTTGCGGTT 
                 
               
               
                   
               
               
                 
                   GATCTATCTCGCGCTGGCGCATATGATCAAATTTCGGGGACACTTCCTCATCGAGGGGGACCTGAACC 
                 
               
               
                   
               
               
                 
                   CAGACAACAGCGATGTCGACAAACTCTTTATCCAACTGGTTCAGACTTACAATCAGCTTTTCGAAGAG 
                 
               
               
                   
               
               
                 
                   AACCCGATCAACGCATCCGGAGTTGACGCCAAAGCAATCCTGAGCGCTAGGCTGTCCAAATCCCGGCG 
                 
               
               
                   
               
               
                 
                   GCTCGAAAACCTCATCGCACAGCTCCCTGGGGAGAAGAAGAACGGCCTGTTTGGTAATCTTATCGCCC 
                 
               
               
                   
               
               
                 
                   TGTCACTCGGGCTGACCCCCAACTTTAAATCTAACTTCGACCTGGCCGAAGATGCCAAGCTTCAACTG 
                 
               
               
                   
               
               
                 
                   AGCAAAGACACCTACGATGATGATCTCGACAATCTGCTGGCCCAGATCGGCGACCAGTACGCAGACCT 
                 
               
               
                   
               
               
                 
                   TTTTTTGGCGGCAAAGAACCTGTCAGACGCCATTCTGCTGAGTGATATTCTGCGAGTGAACACGGAGA 
                 
               
               
                   
               
               
                 
                   TCACCAAAGCtCCGCTGAGCGCTAGTATGATCAAGCGCTATGATGAGCACCACCAAGACTTGACTTTG 
                 
               
               
                   
               
               
                 
                   CTGAAGGCCCTTGTCAGACAGCAACTGCCTGAGAAGTACAAGGAAATTTTCTTCGATCAGTCTAAAAA 
                 
               
               
                   
               
               
                 
                   TGGCTACGCCGGATACATTGACGGCGGAGCAAGCCAGGAGGAATTTTACAAATTTATTAAGCCCATCT 
                 
               
               
                   
               
               
                 
                   TGGAAAAAATGGACGGCACCGAGGAGCTGCTGGTAAAGCTTAACAGAGAAGATCTGTTGCGCAAACAG 
                 
               
               
                   
               
               
                 
                   CGCACTTTCGACAATGGAAGCATCCCCCACCAGATTCACCTGGGCGAACTGCACGCTATCCTCAGGCG 
                 
               
               
                   
               
               
                 
                   GCAAGAGGATTTCTACCCCTTTTTGAAAGATAACAGGGAAAAGATTGAGAAAATCCTCACATTTCGGA 
                 
               
               
                   
               
               
                 
                   TACCCTACTATGTAGGCCCCCTCGCCCGGGGAAATTCCAGATTCGCGTGGATGACTCGCAAATCAGAA 
                 
               
               
                   
               
               
                 
                   GAGACCATCACTCCCTGGAACTTCGAGGAAGTCGTGGATAAGGGGGCCTCTGCCCAGTCCTTCATCGA 
                 
               
               
                   
               
               
                 
                   AAGGATGACTAACTTTGATAAAAATCTGCCTAACGAAAAGGTGCTTCCTAAACACTCTCTGCTGTACG 
                 
               
               
                   
               
               
                 
                   AGTACTTCACAGTTTATAACGAGCTCACCAAGGTCAAATACGTCACAGAAGGGATGAGAAAGCCAGCA 
                 
               
               
                   
               
               
                 TTCCTGTCTGGAGAGCAGAAGAAAGCTATCGTGGACCTCCTCTTCAAGACGAACCGGAAAGTTACCGT 
               
               
                   
               
               
                 
                   GAAACAGCTCAAAGAAGACTATTTCAAAAAGATTGAATGTTTCGACTCTGTTGAAATCAGCGGAGTGG 
                 
               
               
                   
               
               
                 
                   AGGATCGCTTCAACGCATCCCTGGGAACGTATCACGATCTCCTGAAAATCATTAAAGACAAGGACTTC 
                 
               
               
                   
               
               
                 
                   CTGGACAATGAGGAGAACGAGGACATTCTTGAGGACATTGTCCTCACCCTTACGTTGTTTGAAGATAG 
                 
               
               
                   
               
               
                 
                   GGAGATGATTGAAGAACGCTTGAAAACTTACGCTCATCTCTTCGACGACAAAGTCATGAAACAGCTCA 
                 
               
               
                   
               
               
                 
                   AGAGGCGCCGATATACAGGATGGGGGCGGCTGTCAAGAAAACTGATCAATGGGATCCGAGACAAGCAG 
                 
               
               
                   
               
               
                 
                   AGTGGAAAGACAATCCTGGATTTTCTTAAGTCCGATGGATTTGCCAACCGGAACTTCATGCAGTTGAT 
                 
               
               
                   
               
               
                 
                   CCATGATGACTCTCTCACCTTTAAGGAGGACATCCAGAAAGCACAAGTTTCTGGCCAGGGGGACAGTC 
                 
               
               
                   
               
               
                 
                   TTCACGAGCACATCGCTAATCTTGCAGGTAGCCCAGCTATCAAAAAGGGAATACTGCAGACCGTTAAG 
                 
               
               
                   
               
               
                 
                   GTCGTGGATGAACTCGTCAAAGTAATGGGAAGGCATAAGCCCGAGAATATCGTTATCGAGATGGCCCG 
                 
               
               
                   
               
               
                 
                   AGAGAACCAAACTACCCAGAAGGGACAGAAGAACAGTAGGGAAAGGATGAAGAGGATTGAAGAGGGTA 
                 
               
               
                   
               
               
                 
                   TAAAAGAACTGGGGTCCCAAATCCTTAAGGAACACCCAGTTGAAAACACCCAGCTTCAGAATGAGAAG 
                 
               
               
                   
               
               
                 
                   CTCTACCTGTACTACCTGCAGAACGGCAGGGACATGTACGTGGATCAGGAACTGGACATCAATCGGCT 
                 
               
               
                   
               
               
                 
                   CTCCGACTACGACGTGGATGCCATCGTGCCCCAGTCTTTTCTCAAAGATGATTCTATTGATAATAAAG 
                 
               
               
                   
               
               
                 
                   TGTTGACAAGATCCGATAAAAATAGAGGGAAGAGTGATAACGTCCCCTCAGAAGAAGTTGTCAAGAAA 
                 
               
               
                   
               
               
                 
                   ATGAAAAATTATTGGCGGCAGCTGCTGAACGCCAAACTGATCACACAACGGAAGTTCGATAATCTGAC 
                 
               
               
                   
               
               
                 
                   TAAGGCTGAACGAGGTGGCCTGTCTGAGTTGGATAAAGCCGGCTTCATCAAAAGGCAGCTTGTTGAGA 
                 
               
               
                   
               
               
                 
                   CACGCCAGATCACCAAGCACGTGGCCCAAATTCTCGATTCACGCATGAACACCAAGTACGATGAAAAT 
                 
               
               
                   
               
               
                 
                   GACAAACTGATTCGAGAGGTGAAAGTTATTACTCTGAAGTCTAAGCTGGTCTCAGATTTCAGAAAGGA 
                 
               
               
                   
               
               
                 
                   CTTTCAGTTTTATAAGGTGAGAGAGATCAACAATTACCACCATGCGCATGATGCCTACCTGAATGCAG 
                 
               
               
                   
               
               
                 
                   TGGTAGGCACTGCACTTATCAAAAAATATCCCAAGCTTGAATCTGAATTTGTTTACGGAGACTATAAA 
                 
               
               
                   
               
               
                 
                   GTGTACGATGTTAGGAAAATGATCGCAAAGTCTGAGCAGGAAATAGGCAAGGCCACCGCTAAGTACTT 
                 
               
               
                   
               
               
                 
                   CTTTTACAGCAATATTATGAATTTTTTCAAGACCGAGATTACACTGGCCAATGGAGAGATTCGGAAGC 
                 
               
               
                   
               
               
                 
                   GACCACTTATCGAAACAAACGGAGAAACAGGAGAAATCGTGTGGGACAAGGGTAGGGATTTCGCGACA 
                 
               
               
                   
               
               
                 
                   GTCCGGAAGGTCCTGTCCATGCCGCAGGTGAACATCGTTAAAAAGACCGAAGTACAGACCGGAGGCTT 
                 
               
               
                   
               
               
                 
                   CTCCAAGGAAAGTATCCTCCCGAAAAGGAACAGCGACAAGCTGATCGCACGCAAAAAAGATTGGGACC 
                 
               
               
                   
               
               
                 
                   CCAAGAAATACGGCGGATTCGATTCTCCTACAGTCGCTTACAGTGTACTGGTTGTGGCCAAAGTGGAG 
                 
               
               
                   
               
               
                 
                   AAAGGGAAGTCTAAAAAACTCAAAAGCGTCAAGGAACTGCTGGGCATCACAATCATGGAGCGATCAAG 
                 
               
               
                   
               
               
                 
                   CTTCGAAAAAAACCCCATCGACTTTCTCGAGGCGAAAGGATATAAAGAGGTCAAAAAAGACCTCATCA 
                 
               
               
                   
               
               
                 
                   TTAAGCTTCCCAAGTACTCTCTCTTTGAGCTTGAAAACGGCCGGAAACGAATGCTCGCTAGTGCGGGC 
                 
               
               
                   
               
               
                 
                   GAGCTGCAGAAAGGTAACGAGCTGGCACTGCCCTCTAAATACGTTAATTTCTTGTATCTGGCCAGCCA 
                 
               
               
                   
               
               
                 
                   CTATGAAAAGCTCAAAGGGTCTCCCGAAGATAATGAGCAGAAGCAGCTGTTCGTGGAACAACACAAAC 
                 
               
               
                   
               
               
                 
                   ACTACCTTGATGAGATCATCGAGCAAATAAGCGAATTCTCCAAAAGAGTGATCCTCGCCGACGCTAAC 
                 
               
               
                   
               
               
                 
                   CTCGATAAGGTGCTTTCTGCTTACAATAAGCACAGGGATAAGCCCATCAGGGAGCAGGCAGAAAACAT 
                 
               
               
                   
               
               
                 
                   TATCCACTTGTTTACTCTGACCAACTTGGGCGCGCCTGCAGCCTTCAAGTACTTCGACACCACCATAG 
                 
               
               
                   
               
               
                 
                   ACAGAAAGCGGTACACCTCTACAAAGGAGGTCCTGGACGCCACACTGATTCATCAGTCAATTACGGGG 
                 
               
               
                   
               
               
                 
                   CTCTATGAAACAAGAATCGACCTCTCTCAGCTCGGTGGAGACAGCAGGGCTGAC 
                 
               
               
                   
               
               
                 Amino acid sequence for VPH-dCas9 
               
               
                 (corresponding to SEQ ID NO: 69); lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9 
               
               
                 dykdhdgdykdhdidykddddkhv dalddfdldmlgsdalddfdldmlgsdalddfdldmlgsdaldd   
               
               
                 SEQ ID NO: 70 
                   
               
               
                 
                   fdldmlgslpsasvefegsggpsgaisngalalapssapvlaqtmvpssamvplaqppapapvltpgp 
                 
                   
               
               
                   
               
               
                 
                   pqslsapvpkstqagegtlseallhlqfdadedlgallgnstdpgvftdlasydnsefqqllnqgvsm 
                 
               
               
                   
               
               
                 
                   shstaepmlmeypeaitrlvtgsqrppdpaptplqtsglpnqlsgdedfssiadmdfsailsqisssg 
                 
               
               
                   
               
               
                 
                   qggggsqfsvdtsalldlfspsvtvpdmslpdldsslasiqellspqepprppeaensspdsqkqlvh 
                 
               
               
                   
               
               
                   ytaqplflldpgsvdtgsndlpvlfelgegsyfsegdgfaedptislllqseppkakdptvs npkkkr 
               
               
                   
               
               
                 kvgrg MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATR   
               
               
                   
               
               
                 
                   LKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEK 
                 
               
               
                   
               
               
                 
                   YPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEEN 
                 
               
               
                   
               
               
                 
                   PINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLS 
                 
               
               
                   
               
               
                 
                   KDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLL 
                 
               
               
                   
               
               
                 
                   KALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQR 
                 
               
               
                   
               
               
                 
                   TFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEE 
                 
               
               
                   
               
               
                 
                   TITPWNFEEVVDKGASAQSFTERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAE 
                 
               
               
                   
               
               
                 
                   LSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFL 
                 
               
               
                   
               
               
                 
                   DNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQS 
                 
               
               
                   
               
               
                 
                   GKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKV 
                 
               
               
                   
               
               
                 
                   VDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKL 
                 
               
               
                   
               
               
                 
                   YLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKM 
                 
               
               
                   
               
               
                 
                   KNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDEND 
                 
               
               
                   
               
               
                 
                   KLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKV 
                 
               
               
                   
               
               
                 
                   YDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATV 
                 
               
               
                   
               
               
                 
                   RKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEK 
                 
               
               
                   
               
               
                 
                   GKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGE 
                 
               
               
                   
               
               
                 
                   LQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANL 
                 
               
               
                   
               
               
                 
                   DKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGL 
                 
               
               
                 
                   YETRIDLSQLGGDSRAD 
                 
               
               
                   
               
               
                 DNA sequence for dCas9-VPH (in backbone pN/36): 
               
               
                 pNI70; lowercase underlined = VPH; 
               
               
                 capital underlined = dCas9. 
               
               
                 SEQ ID NO: 71 
                   
               
               
                 
                   ATGGACAAGAAGTACAGCATCGGCCTGGCCATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGA 
                 
                   
               
               
                   
               
               
                 
                   CGAGTACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAAGAAGA 
                 
               
               
                   
               
               
                 
                   ACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGCTGAAGAGAACCGCC 
                 
               
               
                   
               
               
                 
                   AGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGC 
                 
               
               
                   
               
               
                 
                   CAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAAGCACG 
                 
               
               
                   
               
               
                 
                   AGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTAC 
                 
               
               
                   
               
               
                 
                   CACCTGAGAAAGAAACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGC 
                 
               
               
                   
               
               
                 
                   CCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGG 
                 
               
               
                   
               
               
                 
                   ACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGC 
                 
               
               
                   
               
               
                 
                   GGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAATCTGATCGC 
                 
               
               
                   
               
               
                 
                   CCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAGCCTGGGCCTGACCC 
                 
               
               
                   
               
               
                 
                   CCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCAAGGACACCTACGAC 
                 
               
               
                   
               
               
                 
                   GACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTTCTGGcCGCCAAGAA 
                 
               
               
                   
               
               
                 
                   CCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGA 
                 
               
               
                   
               
               
                 
                   GcGCCTCTATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGTGcGG 
                 
               
               
                   
               
               
                 
                   CAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGCTACGCCGGCTACAT 
                 
               
               
                   
               
               
                 
                   TGACGGCGGAGCCAGCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCA 
                 
               
               
                   
               
               
                 
                   CCGAGGAACTGCTCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAACGGC 
                 
               
               
                   
               
               
                 
                   AGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAGGAAGATTTTTACCC 
                 
               
               
                   
               
               
                 
                   ATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCCCTAGTAGGTGGGCC 
                 
               
               
                   
               
               
                 
                   CTCTGGCCAGGGGAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGG 
                 
               
               
                   
               
               
                 
                   AACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGGATGACCAACTTCGA 
                 
               
               
                   
               
               
                 
                   TAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTACTTCACCGTGTATA 
                 
               
               
                   
               
               
                 
                   ACGAGCTGACCAAAGTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAG 
                 
               
               
                   
               
               
                 
                   AAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCAGCTGAAAGAGGA 
                 
               
               
                   
               
               
                 
                   CTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCCGGCGTGGAAGATCGGTTCAACGCCT 
                 
               
               
                   
               
               
                 
                   CCCTGGGCACATACCACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAAC 
                 
               
               
                   
               
               
                 
                   GAGGACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCGAGGAACG 
                 
               
               
                   
               
               
                 
                   GCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCGGCGGAGATACACCG 
                 
               
               
                   
               
               
                 
                   GCTGGGGCAGGCTGAGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTG 
                 
               
               
                   
               
               
                 
                   GATTTCCTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACGACAGCCTGAC 
                 
               
               
                   
               
               
                 
                   CTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCACGAGCACATTGCCA 
                 
               
               
                   
               
               
                 
                   ATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGGTGGACGAGCTCGTG 
                 
               
               
                   
               
               
                 
                   AAAGTGATGGGCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCCA 
                 
               
               
                   
               
               
                 
                   GAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAAAGAGCTGGGCAGCC 
                 
               
               
                   
               
               
                 
                   AGATCCTGAAAGAACACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTG 
                 
               
               
                   
               
               
                 
                   CAGAATGGGCGGGATATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGA 
                 
               
               
                   
               
               
                 
                   CGCCATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCTGACCAGAAGCGACA 
                 
               
               
                   
               
               
                 
                   AGAACCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGAAGAACTACTGGCGG 
                 
               
               
                   
               
               
                 
                   CAGCTGLTGAACGCCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGGCGG 
                 
               
               
                   
               
               
                 
                   CCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCGGLAGATCACAAAGC 
                 
               
               
                   
               
               
                 
                   ACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACAAGCTGATCCGGGAA 
                 
               
               
                   
               
               
                 
                   GTGAAAGTGATCACCCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGT 
                 
               
               
                   
               
               
                 
                   GCGCGAGATCAACAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGTGGGAACCGCCCTGA 
                 
               
               
                   
               
               
                 
                   TCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGTACGACGTGCGGAAG 
                 
               
               
                   
               
               
                 
                   ATGATCGCCAAGAGCGAGCAGGAAATCGGCAAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCAT 
                 
               
               
                   
               
               
                 
                   GAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCCTCTGATCGAGACAA 
                 
               
               
                   
               
               
                 
                   ACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGCGGAAAGTGCTGAGC 
                 
               
               
                   
               
               
                 
                   ATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCT 
                 
               
               
                   
               
               
                 
                   GCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCT 
                 
               
               
                   
               
               
                 
                   TCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAA 
                 
               
               
                   
               
               
                 
                   CTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCAT 
                 
               
               
                   
               
               
                 
                   CGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACT 
                 
               
               
                   
               
               
                 
                   CCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAGAAGGGAAAC 
                 
               
               
                   
               
               
                 
                   GAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTATGAGAAGCTGAAGGG 
                 
               
               
                   
               
               
                 
                   CTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTACCTGGACGAGATCA 
                 
               
               
                   
               
               
                 
                   TCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCC 
                 
               
               
                   
               
               
                 
                   GCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCT 
                 
               
               
                   
               
               
                 
                   GACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCA 
                 
               
               
                   
               
               
                 
                   GCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGATC 
                 
               
               
                   
               
               
                   GACCTGTCTCAGCTGGGAGGCGAC aagcgacctgccgccacaaagaagqctggacaggctaagaagaa 
               
               
                   
               
               
                 gaaactggactctggaggatccgactacaaagaccatgacggtgattataaagatcatgacatcgatt 
               
               
                   
               
               
                 acaaggatgacgatgacaagggaggatccaaggagaagagtgcttgtcctaaagatccagccaaacct 
               
               
                   
               
               
                 ccggccaaggcacaagttgtgggatggccaccggtgagatcataccggaagaacgtgatggtttcctg 
               
               
                   
               
               
                 ccaaaaatcaagcggtggcccggaggcggcggcgttcgtgaaggtatcaatggacggagcaccgtact 
               
               
                   
               
               
                 tgaggaaaatcgatttgaggatgtataaaggcggatctggcggctctggaggatccagc gatgcttta   
               
               
                   
               
               
                 
                   gacgattttgacttagatatgcttggttcagacgcgttagacgacttcgacctagacatgttaggctc 
                 
               
               
                   
               
               
                 
                   agatgcattggacgacttcgatttagatatgttgggctccgatgccctagatgactttgatctagata 
                 
               
               
                   
               
               
                 
                   tgctagggtcactacccagcgctagcgtcgagttcgaaggcagcggcgggccttcagggcagatcagc 
                 
               
               
                   
               
               
                 
                   aaccaggccctggctctggcccctagctccgctccagtgctggcccagactatggtgccctctagtgc 
                 
               
               
                   
               
               
                 
                   tatggtgcctctggcccagccacctgctccagcccctgtgctgaccccaggaccaccccagtcactga 
                 
               
               
                   
               
               
                 
                   gcgctccagtgcccaagtctacacaggccggcgaggggactctgagtgaagctctgctgcacctgcag 
                 
               
               
                   
               
               
                 
                   ttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccggagtgttcacagatct 
                 
               
               
                   
               
               
                 
                   ggcctccgtggacaactctgagtttcagcagctgctgaatcagggcgtgtccatgtctcatagtacag 
                 
               
               
                   
               
               
                 
                   ccgaaccaatgctgatggagtaccccgaagccattacccggctggtgaccggcagccagcggcccccc 
                 
               
               
                   
               
               
                 
                   gaccccgctccaactcccctgggaaccageggcctgcctaatgggctatcoggagatgaagacttctc 
                 
               
               
                   
               
               
                 
                   aagcatcgctgatatggactttagtgccctgctgtcacagatttcctctagtgggcagggaggaggtg 
                 
               
               
                   
               
               
                 
                   gaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcccctcggtgaccgtgcccgac 
                 
               
               
                   
               
               
                 
                   atgagcctgcctgaccttgacagcagcctggccagtatccaagagctcctgtctccccaggagccccc 
                 
               
               
                   
               
               
                 
                   caggcctcccgaggcagagaacagcagcccggattcagggaagcagctggtgcactacacagcgcagc 
                 
               
               
                   
               
               
                 
                   cgctgttcctgctggaccccggctccgtggacaccgggagcaacgacctgccggtgctgtttgagctg 
                 
               
               
                   
               
               
                 
                   ggagagggctcctacttctccgaaggggacggcttcgccgaggaccccaccatctccctgctgacagg 
                 
               
               
                   
               
               
                 
                   ctcggagcctcccaaagccaaggaccccactgtctcctga 
                 
               
               
                   
               
               
                 Amino acid sequence fordCas9-VPH 
               
               
                 (corresponding to SEQ ID NO: 71); lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 72 
                   
               
               
                 
                   MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                 
                   
               
               
                   
               
               
                 
                   RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIY 
                 
               
               
                   
               
               
                 
                   HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
                 
               
               
                   
               
               
                 
                   GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
                 
               
               
                   
               
               
                 
                   DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
                 
               
               
                   
               
               
                 
                   QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
                 
               
               
                   
               
               
                 
                   SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
                 
               
               
                   
               
               
                 
                   NFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQ 
                 
               
               
                   
               
               
                 
                   KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
                 
               
               
                   
               
               
                 
                   EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
                 
               
               
                   
               
               
                 
                   DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
                 
               
               
                   
               
               
                 
                   KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
                 
               
               
                   
               
               
                 
                   QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
                 
               
               
                   
               
               
                 
                   QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
                 
               
               
                   
               
               
                 
                   VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
                 
               
               
                   
               
               
                 
                   MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
                 
               
               
                   
               
               
                 
                   MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKK 
                 
               
               
                   
               
               
                 
                   LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
                 
               
               
                   
               
               
                 
                   ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLS 
                 
               
               
                   
               
               
                 
                   AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
                 
               
               
                   
               
               
                   DLSQLGGD krpaatkkagqakkkkldsggsdykdhdgdykdhdidykddddkggskeksacpkdpakp 
               
               
                   
               
               
                 pakaqvvgwppvrsyrknvmvscqkssggpeaaafvkvsmdgapylrkidlrmykggsggsggss dal   
               
               
                   
               
               
                 
                   ddfdldmlgsdalddfdldmlgsdalddfdldmlgsdalddfdldmlgslpsasvefegsggpsgqis 
                 
               
               
                   
               
               
                 
                   nqalalapssapvlaqtmvpssamvplaqppapapvltpgppgslsapvpkstqagegtlseallhlq 
                 
               
               
                   
               
               
                 
                   fdadedlgallgnstdpgvftdlasvdnsefaqllnqgvsmshstaepmlmeypeaitrlvtgsqrpp 
                 
               
               
                   
               
               
                 
                   dpaptplgtsglpnglsgdedfssiadmdfsallsqisssgqggggsgfsvdtsalldlfspsvtvpd 
                 
               
               
                   
               
               
                 
                   mslpdldsslasiqellspqepprppeaensspdsgkqlvhytaqplflldpgsvdtgsndlpvlfel 
                 
               
               
                   
               
               
                 
                   gegsvfsegdgfaedptislitgseppkakdptvs* 
                 
               
               
                   
               
               
                 DNA sequence fordCas9-VPH (in backbone pNH 23): pNI137; 
               
               
                 lowercase underlined = VPH; 
               
               
                 capital underlined = dCas9. 
               
               
                 SEQ ID NO: 73 
                   
               
               
                 atggactacaaagaccatgacggtgattataaagatcatgacatcgattacaaggatgacgatgacaa 
                   
               
               
                   
               
               
                 gcacgttaaccccaagaagaagaggaaggtgggccgcgga ATGGACAAGAAGTACTCCATTGGGCTCG   
               
               
                   
               
               
                 
                   CCATCGGCACAAACAGCGTCGGCTGGGCCGTCATTACGGACGAGTACAAGGTGCCGAGCAAAAAATTC 
                 
               
               
                   
               
               
                 
                   AAAGTTCTGGGCAATACCGATCGCCACAGCATAAAGAAGAACCTCATTGGCGCCCTCCTGTTCGACTC 
                 
               
               
                   
               
               
                 
                   CGGGGAAACCGCCGAAGCCACGCGGCTCAAAAGAACAGCACGGCGCAGATATACCCGCAGAAAGAATC 
                 
               
               
                   
               
               
                 
                   GGATCTGCTACCTGCAGGAGATCTTTAGTAATGAGATGGCTAAGGTGGATGACTCTTTCTTCCATAGG 
                 
               
               
                   
               
               
                 
                   CTGGAGGAGTCCTTTTTGGTGGAGGAGGATAAAAAGCACGAGCGCCACCCAATCTTTGGCAATATCGT 
                 
               
               
                   
               
               
                 
                   GGACGAGGTGGCGTACCATGAAAAGTACCCAACCATATATCATCTGAGGAAGAAGCTTGTAGACAGTA 
                 
               
               
                   
               
               
                 
                   CTGATAAGGCTGACTTGCGGTTGATCTATCTCGCGCTGGCGCATATGATCAAATTTCGGGGACACTTC 
                 
               
               
                   
               
               
                 
                   CTCATCGAGGGGGACCTGAACCCAGACAACAGCGATGTCGACAAACTCTTTATCCAACTGGTTCAGAC 
                 
               
               
                   
               
               
                 
                   TTACAATCAGCTTTTCGAAGAGAACCCGATCAACGCATCCGGAGTTGACGCCAAAGCAATCCTGAGCG 
                 
               
               
                   
               
               
                 
                   CTAGGCTGTCCAAATCCCGGCGGCTCGAAAACCTCATCGCACAGCTCCCTGGGGAGAAGAAGAACGGC 
                 
               
               
                   
               
               
                 
                   CTGTTTGGTAATCTTATCGCCCTGTCACTCGGGCTGACCCCCAACTTTAAATCTAACTTCGACCTGGC 
                 
               
               
                   
               
               
                 
                   CGAAGATGCCAAGCTTCAACTGAGCAAAGACACCTACGATGATGATCTCGACAATCTGCTGGCCCAGA 
                 
               
               
                   
               
               
                 
                   TCGGCGACCAGTACGCAGACCTTTTTTTGGCGGCAAAGAACCTGTCAGACGCCATTCTGCTGAGTGAT 
                 
               
               
                   
               
               
                 
                   ATTCTGCGAGTGAACACGGAGATCACCAAAGCTCCGCTGAGCGCTAGTATGATCAAGCGCTATGATGA 
                 
               
               
                   
               
               
                 
                   GCACCACCAAGACTTGACTTTGCTGAAGGCCCTTGTCAGACAGCAACTGCCTGAGAAGTACAAGGAAA 
                 
               
               
                   
               
               
                 
                   TTTTCTTCGATCAGTCTAAAAATGGCTACGCCGGATACATTGACGGCGGAGCAAGCCAGGAGGAATTT 
                 
               
               
                   
               
               
                 
                   TACAAATTTATTAAGCCCATCTTGGAAAAAATGGACGGCACCGAGGAGCTGCTGGTAAAGCTTAACAG 
                 
               
               
                   
               
               
                 
                   AGAAGATCTGTTGCGCAAACAGCGCACTTTCGACAATGGAAGCATCCCCCACCAGATTCACCTGGGCG 
                 
               
               
                   
               
               
                 
                   AACTGCACGCTATCCTCAGGCGGCAAGAGGATTTCTACCCCTTTTTGAAAGATAACAGGGAAAAGATT 
                 
               
               
                   
               
               
                 
                   GAGAAAATCCTCACATTTCGGATACCCTACTATGTAGGCCCCCTCGCCCGGGGAAATTCCAGATTCGC 
                 
               
               
                   
               
               
                 
                   GTGGATGACTCGCAAATCAGAAGAGACCATCACTCCCTGGAACTTCGAGGAAGTCGTGGATAAGGGGG 
                 
               
               
                   
               
               
                 
                   CCTCTGCCCAGTCCTTCATCGAAAGGATGACTAACTTTGATAAAAATCTGCCTAACGAAAAGGTGCTT 
                 
               
               
                   
               
               
                 
                   CCTAAACACTCTCTGCTGTACGAGTACTTCACAGTTTATAACGAGCTCACCAAGGTCAAATACGTCAC 
                 
               
               
                   
               
               
                 
                   AGAAGGGATGAGAAAGCCAGCATTCCTGTCTGGAGAGCAGAAGAAAGCTATCGTGGACCTCCTCTTCA 
                 
               
               
                   
               
               
                 
                   AGACGAACCGGAAAGTTACCGTGAAACAGCTCAAAGAAGACTATTTCAAAAAGATTGAATGTTTCGAC 
                 
               
               
                   
               
               
                 
                   TCTGTTGAAATCAGCGGAGTGGAGGATCGCTTCAACGCATCCCTGGGAACGTATCACGATCTCCTGAA 
                 
               
               
                   
               
               
                 
                   AATCATTAAAGACAAGGACTTCCTGGACAATGAGGAGAACGAGGACATTCTTGAGGACATTGTCCTCA 
                 
               
               
                   
               
               
                 
                   CCCTTACGTTGTTTGAAGATAGGGAGATGATTGAAGAACGCTTGAAAACTTACGCTCATCTCTTCGAC 
                 
               
               
                   
               
               
                 
                   GACAAAGTCATGAAACAGCTCAAGAGGCGCCGATATACAGGATGGGGGCGGCTGTCAAGAAAACTGAT 
                 
               
               
                   
               
               
                 
                   CAATGGGATCCGAGACAAGCAGAGTGGAAAGACAATCCTGGATTTTCTTAAGTCCGATGGATTTGCCA 
                 
               
               
                   
               
               
                 
                   ACCGGAACTTCATGCAGTTGATCCATGATGACTCTCTCACCTTTAAGGAGGACATCCAGAAAGCACAA 
                 
               
               
                   
               
               
                 
                   GTTTCTGGCCAGGGGGACAGTCTTCACGAGCACATCGCTAATCTTGCAGGTAGCCCAGCTATCAAAAA 
                 
               
               
                   
               
               
                 
                   GGGAATACTGCAGACCGTTAAGGTCGTGGATGAACTCGTCAAAGTAATGGGAAGGCATAAGCCCGAGA 
                 
               
               
                   
               
               
                 
                   ATATCGTTATCGAGATGGCCCGAGAGAACCAAACTACCCAGAAGGGACAGAAGAACAGTAGGGAAAGG 
                 
               
               
                   
               
               
                 
                   ATGAAGAGGATTGAAGAGGGTATAAAAGAACTGGGGTCCCAAATCCTTAAGGAACACCCAGTTGAAAA 
                 
               
               
                   
               
               
                 
                   CACCCAGCTTCAGAATGAGAAGCTCTACCTGTACTACCTGCAGAACGGCAGGGACATGTACGTGGATC 
                 
               
               
                   
               
               
                 
                   AGGAACTGGACATCAATCGGCTCTCCGACTACGACGTGGATGCCATCGTGCCCCAGTCTTTTCTCAAA 
                 
               
               
                   
               
               
                 
                   GATGATTCTATTGATAATAAAGTGTTGACAAGATCCGATAAAAATAGAGGGAAGAGTGATAACGTCCC 
                 
               
               
                   
               
               
                 
                   CTCAGAAGAAGTTGTCAAGAAAATGAAAAATTATTGGCGGCAGCTGCTGAACGCCAAACTGATCACAC 
                 
               
               
                   
               
               
                 
                   AACGGAAGTTCGATAATCTGACTAAGGCTGAACGAGGTGGCCTGTCTGAGTTGGATAAAGCCGGCTTC 
                 
               
               
                   
               
               
                 
                   ATCAAAAGGCAGCTTGTTGAGACACGCCAGATCACCAAGCACGTGGCCCAAATTCTCGATTCACGCAT 
                 
               
               
                   
               
               
                 
                   GAACACCAAGTACGATGAAAATGACAAACTGATTCGAGAGGTGAAAGTTATTACTCTGAAGTCTAAGC 
                 
               
               
                   
               
               
                 
                   TGGTCTCAGATTTCAGAAAGGACTTTCAGTTTTATAAGGTGAGAGAGATCAACAATTACCACCATGCG 
                 
               
               
                   
               
               
                 
                   CATGATGCCTACCTGAATGCAGTGGTAGGCACTGCACTTATCAAAAAATATCCCAAGCTTGAATCTGA 
                 
               
               
                   
               
               
                 
                   ATTTGTTTACGGAGACTATAAAGTGTACGATGTTAGGAAAATGATCGCAAAGTCTGAGCAGGAAATAG 
                 
               
               
                   
               
               
                 
                   GCAAGGCCACCGCTAAGTACTTCTTTTACAGCAATATTATGAATTTTTTCAAGACCGAGATTACACTG 
                 
               
               
                   
               
               
                 
                   GCCAATGGAGAGATTCGGAAGCGACCACTTATCGAAACAAACGGAGAAACAGGAGAAATCGTGTGGGA 
                 
               
               
                   
               
               
                 
                   CAAGGGTAGGGATTTCGCGACAGTCCGGAAGGTCCTGTCCATGCCGCAGGTGAACATCGTTAAAAAGA 
                 
               
               
                   
               
               
                 
                   CCGAAGTACAGACCGGAGGCTTCTCCAAGGAAAGTATCCTCCCGAAAAGGAACAGCGACAAGCTGATC 
                 
               
               
                   
               
               
                 
                   GCACGCAAAAAAGATTGGGACCCCAAGAAATACGGCGGATTCGATTCTCCTACAGTCGCTTACAGTGT 
                 
               
               
                   
               
               
                 
                   ACTGGTTGTGGCCAAAGTGGAGAAAGGGAAGTCTAAAAAACTCAAAAGCGTCAAGGAACTGCTGGGCA 
                 
               
               
                   
               
               
                 
                   TCACAATCATGGAGCGATCAAGCTTCGAAAAAAACCCCATCGACTTTCTCGAGGCGAAAGGATATAAA 
                 
               
               
                   
               
               
                 
                   GAGGTCAAAAAAGACCTCATCATTAAGCTTCCCAAGTACTCTCTCTTTGAGCTTGAAAACGGCCGGAA 
                 
               
               
                   
               
               
                 
                   ACGAATGCTCGCTAGTGCGGGCGAGCTGCAGAAAGGTAACGAGCTGGCACTGCCCTCTAAATACGTTA 
                 
               
               
                   
               
               
                 
                   ATTTCTTGTATCTGGCCAGCCACTATGAAAAGCTCAAAGGGTCTCCCGAAGATAATGAGCAGAAGCAG 
                 
               
               
                   
               
               
                 
                   CTGTTCGTGGAACAACACAAACACTACCTTGATGAGATCATCGAGCAAATAAGCGAATTCTCCAAAAG 
                 
               
               
                   
               
               
                 
                   AGTGATCCTCGCCGACGCTAACCTCGATAAGGTGCTTTCTGCTTACAATAAGCACAGGGATAAGCCCA 
                 
               
               
                   
               
               
                 
                   TCAGGGAGCAGGCAGAAAACATTATCCACTTGTTTACTCTGACCAACTTGGGCGCGCCTGCAGCCTTC 
                 
               
               
                   
               
               
                 
                   AAGTACTTCGACACCACCATAGACAGAAAGCGGTACACCTCTACAAAGGAGGTCCTGGACGCCACACT 
                 
               
               
                   
               
               
                   GATTCATCAGTCAATTACGGGGCTCTATGAAACAAGAATCGACCTCTCTCAGCTCGGTGGAGAC agca 
               
               
                   
               
               
                 gggctgaccccaagaagaagaggaaggtggctagc gatgctttagacgattttgacttagatatgctt   
               
               
                   
               
               
                 
                   ggttcagacgcgttagacgacttcgacctagacatgttaggctcagatgcattggacgacttcgattt 
                 
               
               
                   
               
               
                 
                   agatatgttgggctccgatgccctagatgactttgatetagatatgctagggtcactacccagcgcca 
                 
               
               
                   
               
               
                 
                   gcgtcgagttcgaaggcagcggcgggccttcagggcagatcagcaaccaggccctggctctggcccct 
                 
               
               
                   
               
               
                 
                   agctccgctccagtgctggcccagactatggtgccctctagtgctatggtgcctctggcccagccacc 
                 
               
               
                   
               
               
                 
                   tgctccagcccctatgctgaccccaggaccaccccagtcactgagcgccccagtgcccaagtctacac 
                 
               
               
                   
               
               
                 
                   aggccggcgaggggactctgagtgaagctctgctgcacctgcagttcgacgctgatgaggacctggga 
                 
               
               
                   
               
               
                 
                   gctctgctggggaacagcaccgatcccggagtgttcacagatctggcctccgtggacaactctgagtt 
                 
               
               
                   
               
               
                 
                   tcagcagctgctgaatcagggcgtgtccatgtctcatagtacagccgaaccaatgctgatggagtacc 
                 
               
               
                   
               
               
                 
                   ccgaagccattacccggctggtgaccggcagccagcggccccccgaccccgctccaactcccctggga 
                 
               
               
                   
               
               
                 
                   accagcggcctgcctaatgggctgtccggagatgaagacttctcaagcatcgctgatatggactttag 
                 
               
               
                   
               
               
                 
                   tgccctgctgtcacagatttcctctagtgggcagggaggaggtggaagcggcttcagcgtggacacca 
                 
               
               
                   
               
               
                 
                   gtgccctgctggacctgttcagcccctcggtgaccgtgcccgacatgagcctgcctgaccttgacagc 
                 
               
               
                   
               
               
                 
                   agcctggccagtatccaagagctcctgtctccccaggagccccccaggcctcccgaggcagagaacag 
                 
               
               
                   
               
               
                 cagcccggattcagggaagcagctggtgcactacacagcgcagccgctgttcctgctggaccccggct 
               
               
                   
               
               
                 
                   ccgtggacaccgggagcaacgacctgccggtgctgtttgagctgggagagggctcctacttctccgaa 
                 
               
               
                   
               
               
                 
                   ggggacggcttcgccgaggaccccaccatctccctgctgacaggctcggagcctcccaaagccaagga 
                 
               
               
                   
               
               
                 
                   ccccactgtctcc 
                 
               
               
                   
               
               
                 Amino acid sequence fordCas9-VPH 
               
               
                 (corresponding to SEQ ID NO: 73); lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 74 
                   
               
               
                 dykdhdgaykdhdidykaddddkhvnpkkkrkvgrg MDKKYSIGLAIGTNSVGWAVTDEYKVPSKKFK   
                   
               
               
                   
               
               
                 
                   VLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRL 
                 
               
               
                   
               
               
                 
                   EESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTTYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFL 
                 
               
               
                   
               
               
                 
                   IEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGL 
                 
               
               
                   
               
               
                 
                   FGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDI 
                 
               
               
                   
               
               
                 
                   LRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFY 
                 
               
               
                   
               
               
                 
                   KFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIE 
                 
               
               
                   
               
               
                 
                   KILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLP 
                 
               
               
                   
               
               
                 
                   KHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS 
                 
               
               
                   
               
               
                 
                   VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDD 
                 
               
               
                   
               
               
                 
                   KVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNEMQLIHDDSLTFKEDIQKAQV 
                 
               
               
                   
               
               
                 
                   SGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENTVIEMARENQTTQKGQKNSRERM 
                 
               
               
                   
               
               
                 
                   KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKD 
                 
               
               
                   
               
               
                 
                   DSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFI 
                 
               
               
                   
               
               
                 
                   KRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH 
                 
               
               
                   
               
               
                 
                   DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLA 
                 
               
               
                   
               
               
                 
                   NGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIA 
                 
               
               
                   
               
               
                 
                   RKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKE 
                 
               
               
                   
               
               
                 
                   VKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQL 
                 
               
               
                   
               
               
                 
                   FVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFK 
                 
               
               
                   
               
               
                   YFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD sradpkkkrkvas dalddfdldmlq   
               
               
                   
               
               
                 
                   sdalddfdldmlqsdalddfdldmlqsdalddfdldmlgslpsasvefegsggpsgqisnqalalaps 
                 
               
               
                   
               
               
                 
                   sapvlagtmvpssamvplaqppapapvltpgppqslsapvpkstqagegtlseallhlqfdadedlga 
                 
               
               
                   
               
               
                 
                   llgnstdpgvftdlasvdnsefqqllnggvsmshstaepmlmeypeaitrlvtgsqrppdpaptplgt 
                 
               
               
                   
               
               
                 
                   sglpnglsgdedfssiadmdfsallsqisssgqggggsgfsvdtsalldlfspsvtvpdmslpdldss 
                 
               
               
                   
               
               
                 
                   lasiqellspgepprppeaensspdsqkqlvhytaqplflldpgsvdtgsndlpvlfelgeqsyfseg 
                 
               
               
                   
               
               
                 
                   dgfaedptislltqseppkakdptvs 
                 
               
               
                   
               
               
                 DNA sequence for VPH-dCas9-VPH (in backbone pNI36): pNH 15; lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 75 
                   
               
               
                 
                   atggatgctttagacgattttgacttagatatgcttggttcagacgcgttagacgacttcgacctaga 
                 
                   
               
               
                   
               
               
                 
                   catgttaggctcagatgcattggacgacttcgatttagatatgttgggctccgatgccctagatgact 
                 
               
               
                   
               
               
                 
                   ttgatctagatatgctagggtcactacccagcgccagcgtcgagttcgaaggcagcggcgggccttca 
                 
               
               
                   
               
               
                 
                   gggcagatcagcaaccaggccctggctctggcccctagctccgctccagtgctggcccagactatggt 
                 
               
               
                   
               
               
                 
                   gccctctagtgctatggtgcctctggcccagccacctgctccagcccctgtgctgaccccaggaccac 
                 
               
               
                   
               
               
                 
                   cccagtcactgagcgccccagtgcccaagtctacacaggccggcgaggggactctgagtgaagctctg 
                 
               
               
                   
               
               
                 
                   ctgcacctgcagttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccggagt 
                 
               
               
                   
               
               
                 
                   gttcacagatctggcctccgtggacaactctgagtttcagcagctgctgaatcagggcgtgtccatgt 
                 
               
               
                   
               
               
                 
                   ctcatagtacagccgaaccaatgctgatggagtaccccgaagccattacccggctggtgaccggcagc 
                 
               
               
                   
               
               
                 
                   cagcggccccccgaccccgctccaactcccctgggaaccagcggcctgcctaatgggctgtccggaga 
                 
               
               
                   
               
               
                 
                   tgaagacttctcaagcatcgctgatatggactttagtgccctgctgtcacagatttcctctagtgggc 
                 
               
               
                   
               
               
                 
                   agggaggaggtggaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcccctcggtg 
                 
               
               
                   
               
               
                 
                   accgtgcccgacatgagcctgcctgaccttgacagcagcctggccagtatccaagagctcctgtctcc 
                 
               
               
                   
               
               
                 
                   ccaggagccccccaggcctcccgaggcagagaacagcagcccggattcagggaagcagctggtgcact 
                 
               
               
                   
               
               
                 
                   acacagcgcagccgctgttcctgctggaccccggctccgtggacaccgggagcascgacctgccggtg 
                 
               
               
                   
               
               
                 
                   ctgtttgagctgggagagggctcctacttctccgaaggggacggcttcgccgaggaccccaccatctc 
                 
               
               
                   
               
               
                   cctgctgacaggctcggagcctcccaaagccaaggaccccactgtctcc ggctctggaggatctggcg 
               
               
                   
               
               
                 gctctagcgccacc ATGGACAAGAAGTACAGCATCGGCCTGGCCATCGGCACCAACTCTGTGGGCTGG   
               
               
                   
               
               
                 
                   GCCGTGATCACCGACGAGTACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCA 
                 
               
               
                   
               
               
                 
                   CAGCATCAAGAAGAACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGC 
                 
               
               
                   
               
               
                 
                   TGAAGAGAACCGCCAGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTC 
                 
               
               
                   
               
               
                 
                   AGCAACGAGATGGCCAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGA 
                 
               
               
                   
               
               
                 
                   GGATAAGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGT 
                 
               
               
                   
               
               
                 
                   ACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATC 
                 
               
               
                   
               
               
                 
                   TATCTGGCCCTGGCCCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGA 
                 
               
               
                   
               
               
                 
                   CAACAGCGACGTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACC 
                 
               
               
                   
               
               
                 
                   CCATCAACGCCAGCGGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTG 
                 
               
               
                   
               
               
                 
                   GAAAATCTGATCGCCCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAG 
                 
               
               
                   
               
               
                 
                   CCTGGGCCTGACCCCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCA 
                 
               
               
                   
               
               
                 
                   AGGACACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTT 
                 
               
               
                   
               
               
                 CTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCAC 
               
               
                   
               
               
                 
                   CAAGGCCCCCCTGAGCGCCTGTATGATCAAGAGATACGACGAGCAGGAGGAGGACCTGACCCTGCTGA 
                 
               
               
                   
               
               
                 
                   AAGCTCTCGTGCGGCAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGC 
                 
               
               
                   
               
               
                 
                   TACGCCGGCTACATTGACGGCGGAGCCAGCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGA 
                 
               
               
                   
               
               
                 
                   AAAGATGGACGGCACCGAGGAACTGCTCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGA 
                 
               
               
                   
               
               
                 
                   CCTTCGACAACGGCAGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAG 
                 
               
               
                   
               
               
                 
                   GAAGATTTTTACCCATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCC 
                 
               
               
                   
               
               
                 
                   CTACTACGTGGGCCCTCTGGCCAGGGGAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAA 
                 
               
               
                   
               
               
                 
                   CCATCACCCCCTGGAACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGG 
                 
               
               
                   
               
               
                 
                   ATGACCAACTTCGATAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTA 
                 
               
               
                   
               
               
                 
                   CTTCACCGTGTATAACGAGCTGACCAAAGTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCC 
                 
               
               
                   
               
               
                 
                   TGAGCGGCGAGCAGAAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAG 
                 
               
               
                   
               
               
                 
                   CAGCTGAAAGAGGACTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCCGGCGTGGAAGA 
                 
               
               
                   
               
               
                 
                   TCGGTTCAACGCCTCCCTGGGCACATACCACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGG 
                 
               
               
                   
               
               
                 
                   ACAATGAGGAAAACGAGGACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAG 
                 
               
               
                   
               
               
                 
                   ATGATCGAGGAACGGCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCG 
                 
               
               
                   
               
               
                 
                   GCGGAGATACACCGGCTGGGGCAGGCTGAGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCG 
                 
               
               
                   
               
               
                 
                   GCAAGACAATCCTGGATTTCCTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCAC 
                 
               
               
                   
               
               
                 
                   GACGACAGCCTGACCTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCA 
                 
               
               
                   
               
               
                 
                   CGAGCACATTGCCAATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGG 
                 
               
               
                   
               
               
                 
                   TGGACGAGCTCGTGAAAGTGATGGGCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAG 
                 
               
               
                   
               
               
                 
                   AACCAGACCACCCAGAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAA 
                 
               
               
                   
               
               
                 
                   AGAGCTGGGCAGCCAGATCCTGAAAGAACACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGT 
                 
               
               
                   
               
               
                 
                   ACCTGTACTACCTGCAGAATGGGCGGGATATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCC 
                 
               
               
                   
               
               
                 
                   GACTACGATGTGGACGCCATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCT 
                 
               
               
                   
               
               
                 
                   GACCAGAAGCGACAAGAACCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGA 
                 
               
               
                   
               
               
                 
                   AGAACTACTGGCGGCAGCTGCTGAACGCCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAG 
                 
               
               
                   
               
               
                 
                   GCCGAGAGAGGCGGCCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCG 
                 
               
               
                   
               
               
                 
                   GCAGATCACAAAGCACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACA 
                 
               
               
                   
               
               
                 
                   AGCTGATCCGGGAAGTGAAAGTGATCACCCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTC 
                 
               
               
                   
               
               
                 
                   CAGTTTTACAAAGTGCGCGAGATCAACAATACCACCACGCCCACGACGCCTACCTAGAACGCCGTCGT 
                 
               
               
                   
               
               
                 
                   GGGAACCGCCCTGATCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGT 
                 
               
               
                   
               
               
                 
                   ACGACGTGCGGAAGATGATCGCCAAGAGCGAGCAGGAAATCGGCAAGGCTACCGCCAAGTACTTCTTC 
                 
               
               
                   
               
               
                 
                   TACAGCAACATCATGAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCC 
                 
               
               
                   
               
               
                 
                   TCTGATCGAGACAAACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGC 
                 
               
               
                   
               
               
                 
                   GGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGC 
                 
               
               
                   
               
               
                 
                   AAAGAGTCTATCCTGCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAA 
                 
               
               
                   
               
               
                 
                   GAAGTACGGCGGCTTCGAAAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGG 
                 
               
               
                   
               
               
                 
                   GCAAGTCCAAGAAACTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTC 
                 
               
               
                   
               
               
                 
                   GAGAAGAATCCCATCGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAA 
                 
               
               
                   
               
               
                 
                   GCTGCCTAAGTACTCCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAAC 
                 
               
               
                   
               
               
                 
                   TGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTAT 
                 
               
               
                   
               
               
                 
                   GAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTA 
                 
               
               
                   
               
               
                 
                   CCTGGACGAGATCATCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGG 
                 
               
               
                   
               
               
                 
                   ACAAAGTGCTGTCCGCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATC 
                 
               
               
                   
               
               
                 
                   CACCTGTTTACCCTGACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCG 
                 
               
               
                   
               
               
                 
                   GAAGAGGTACACCAGCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGT 
                 
               
               
                   
               
               
                   ACGAGACACGGATCGACCTGTCTCAGCTGGGAGGCGAC aagcgacctgccgccacaaagaaggctgga 
               
               
                   
               
               
                 caggctaagaagaagaaactggactctggaggatccgactacaaagaccatgacggtgattataaaga 
               
               
                   
               
               
                 tcatgacatcgattacaaggatgacgatgacaagggaggatccaaggagaagagtgcttgtcctaaag 
               
               
                   
               
               
                 atccagccaaacctccggccaaggcacaagttgtgggatggccaccggtgagatcataccggaagaac 
               
               
                   
               
               
                 gtgatggtttcctgccaaaaatcaagcggtggcccggaggcggcggcgttcgtgaaggtatcaatgga 
               
               
                   
               
               
                 cggagcaccgtacttgaggaaaatcgatttgaggatgtataaaggcggatctggcggctctggaggat 
               
               
                   
               
               
                 ccagc gatgctttagacgattttgacttagatatgcttggttcagacgcgttagacgacttcgaccta   
               
               
                   
               
               
                 
                   gacatgttaggctcagatgcattggacgacttcgatttagatatgttgggctccgatgccctagatga 
                 
               
               
                   
               
               
                 
                   ctttgatctagatatgctagggtcactacccagcgctagcgtcgagttcgaaggcagcggcgggcctt 
                 
               
               
                   
               
               
                 
                   cagggcagatcagcaaccaggccctggctctggcccctagctccgctccagtgctggcccagactatg 
                 
               
               
                   
               
               
                 
                   gtgccctctagtgctatggtgcctctggcccagccacctgctccagcccctgtgctgaccccaggacc 
                 
               
               
                   
               
               
                 
                   accccagtcactgagcgctccagtgcccaagtctacacaggccggcgaggggactctgagtgaagctc 
                 
               
               
                   
               
               
                 
                   tgctgcacctgcagttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccgga 
                 
               
               
                   
               
               
                 
                   gtgttcacagatctggcctccgtggacaactctgagtttcagcagctgctgaatcagggcgtgtccat 
                 
               
               
                   
               
               
                 
                   gtctcatagtacagccgaaccaatgctgatggagtaccccgaagccattacccggctggtgaccggca 
                 
               
               
                   
               
               
                 
                   gccagcggccccccgaccccgctccaactcccctgggaaccagcggcctgcctaatgggctgtccgga 
                 
               
               
                   
               
               
                 
                   gatgaagacttctcaagcatogctgatatggactttagtgccctgctgtcacagatttcctctagtgg 
                 
               
               
                   
               
               
                 
                   gcagggaggaggtggaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcccctcgg 
                 
               
               
                   
               
               
                 
                   tgaccgtgcccgacatgagcctgcctgaccttgacagcagcctggccagtatccaagagctcctgtct 
                 
               
               
                   
               
               
                 
                   ccccaggagccccccaggcctcccgaggcagagaacagcagcccggattcagggaagcagctggtgca 
                 
               
               
                   
               
               
                 
                   ctacacagcgcagccgctgttcctgctggaccccggctccgtggacaccgggagcaacgacctgccgg 
                 
               
               
                   
               
               
                 
                   tgctgtttgagctgggagagggctcctacttctccgaaggggacggcttcgccgaggaccccaccatc 
                 
               
               
                   
               
               
                 
                   tccctgctgacaggctcggagcctcccaaagccaaggaccccactgtctcctga 
                 
               
               
                   
               
               
                 Amino acid sequence for VPH-dCas9-VPH 
               
               
                 (corresponding to SEQ ID NO: 75); lowercase 
               
               
                 underlined = VPH; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 76 
                   
               
               
                 
                   dalddfdldmlgsdalddfdldmlgsdalddfdldmlgsdalddfaldmlgslpsasvefegsggpsg 
                 
                   
               
               
                   
               
               
                 
                   qisnqalalapssapvlaqtmvpssamvplaqppapapvltpgppqslsapvpkstqagegtlseall 
                 
               
               
                   
               
               
                 
                   hlqfdadedlgallgnstdpgvftdlasvdnsefqqllnqgvsmshstaepmlmeypeaitrlvtgsq 
                 
               
               
                   
               
               
                 
                   rppdpaptplgtsqlpnglsqdedfssiadmdfsallsqisssgqggqqsgfsvdtsalldlfspsvt 
                 
               
               
                   
               
               
                 
                   vpdmslpdldsslasiqellspqepprppeaensspdsgkqlvhytaqplflldpgsvdtgsndlpvl 
                 
               
               
                   
               
               
                   felgegsyfsegdqfaedptislltgseppkakdptvs gsggsggssat MDKKYSIGLAIGTNSVGWA   
               
               
                   
               
               
                 
                   VITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFS 
                 
               
               
                   
               
               
                 
                   NEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIY 
                 
               
               
                   
               
               
                 
                   LAIAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE 
                 
               
               
                   
               
               
                 
                   NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFL 
                 
               
               
                   
               
               
                 
                   AAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY 
                 
               
               
                   
               
               
                 
                   AGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQE 
                 
               
               
                   
               
               
                 
                   DFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERM 
                 
               
               
                   
               
               
                 
                   TNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQ 
                 
               
               
                   
               
               
                 
                   LKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREM 
                 
               
               
                   
               
               
                 
                   IEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD 
                 
               
               
                   
               
               
                 
                   DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGLLQTVKVVDELVKVMGRHKPENIVIEMAREN 
                 
               
               
                   
               
               
                 
                   QTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSD 
                 
               
               
                   
               
               
                 
                   YDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKA 
                 
               
               
                   
               
               
                 
                   ERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDERKDEQ 
                 
               
               
                   
               
               
                 
                   FYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRRMIAKSEQEIGKATAKYFFY 
                 
               
               
                   
               
               
                 
                   SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK 
                 
               
               
                   
               
               
                 
                   ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFE 
                 
               
               
                   
               
               
                 
                   KNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYE 
                 
               
               
                   
               
               
                 
                   KLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVTLADANLDKVLSAYNKHRDKPIREQAENIIH 
                 
               
               
                   
               
               
                   LFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD krpaatkkagq 
               
               
                   
               
               
                 akkkkldsggsdykdhdgdykdhdidykddddkggskeksacpkdpakppakaqvvgwppvrsyrknv 
               
               
                   
               
               
                 mvscqkssqqpeaaafvkvsmdgapylrkidirmykgqsagsggssdalddfdldmlgsdalddfdid 
               
               
                   
               
               
                 mlgsdalddfdldmlgsdalddfdldmlgslpsasvefegsggpsgqisnqalalapssapvlaqtmv 
               
               
                   
               
               
                 pssamvplaqppapapvltpgppgslsapypkstgagegtlseallhlqfdadedlgallgnstdpqv 
               
               
                   
               
               
                 ftdlasvansefqqllnqgvsmshstaepmlmeypeaitrivtgsqrppdpaptplgtsqlpnglsqd 
               
               
                   
               
               
                 edfssiadmdfsailsqisssgqggggsgfsvdtsalldlfspsvtvpdmslpdldsslasiqellsp 
               
               
                   
               
               
                 qepprppeaensspdsgkqlvhytaqplflldpgsvdtgsndlpvlfelgegsyfsegdqfaedptis 
               
               
                   
               
               
                 lllqseppkakdptvs* 
               
               
                   
               
               
                 DNA sequence fordCas9-VPR (in backbone pNI3S): 
               
               
                 pNi47; lowercase underlined = VPR; 
               
               
                 capital underlined = dCas9. 
               
               
                 SEQ ID NO: 77 
                   
               
               
                 
                   ATGGACAAGAAGTACAGCATCGGCCTGGCCATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGA 
                 
                   
               
               
                   
               
               
                 
                   CGAGTACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAAGAAGA 
                 
               
               
                   
               
               
                 
                   ACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGCTGAAGAGAACCGCC 
                 
               
               
                   
               
               
                 
                   AGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGC 
                 
               
               
                   
               
               
                 
                   CAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAAGCACG 
                 
               
               
                   
               
               
                 
                   AGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTAC 
                 
               
               
                   
               
               
                 
                   CACCTGAGAAAGAAACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGC 
                 
               
               
                   
               
               
                 
                   CCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGG 
                 
               
               
                   
               
               
                 
                   ACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGC 
                 
               
               
                   
               
               
                 
                   GGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAATCTGATCGC 
                 
               
               
                   
               
               
                 
                   CCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAGCCTGGGCCTGACCC 
                 
               
               
                   
               
               
                 
                   CCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCAAGGACACCTACGAC 
                 
               
               
                   
               
               
                 
                   GACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTTCTGGCCGCCAAGAA 
                 
               
               
                   
               
               
                 
                   CCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGA 
                 
               
               
                   
               
               
                 
                   GCGCCTCTATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGTGCGG 
                 
               
               
                   
               
               
                 
                   CAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGCTACGCCGGCTACAT 
                 
               
               
                   
               
               
                 
                   TGACGGCGGAGCCAGCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCA 
                 
               
               
                   
               
               
                 
                   CCGAGGAACTGCTCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAACGGC 
                 
               
               
                   
               
               
                 
                   AGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAGGAAGATTTTTACCC 
                 
               
               
                   
               
               
                 
                   ATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCCCTACTACGTGGGCC 
                 
               
               
                   
               
               
                 
                   CTCTGGCCAGGGGAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGG 
                 
               
               
                   
               
               
                 AACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGGATGACCAACTTCGA 
               
               
                   
               
               
                 
                   TAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTACTTCACCGTGTATA 
                 
               
               
                   
               
               
                 
                   ACGAGCTGACCAAAGTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAG 
                 
               
               
                   
               
               
                 
                   AAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCAGCTGAAAGAGGA 
                 
               
               
                   
               
               
                 
                   CTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCCGGCGTGGAAGATCGGTTCAACGCCT 
                 
               
               
                   
               
               
                 
                   CCCTGGGCACATACCACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAAC 
                 
               
               
                   
               
               
                 
                   GAGGACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCGAGGAACG 
                 
               
               
                   
               
               
                 
                   GCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCGGCGGAGATACACCG 
                 
               
               
                   
               
               
                 
                   GCTGGGGCAGGCTGAGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTG 
                 
               
               
                   
               
               
                 
                   GATTTCCTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATACAGCTGATCCACGACGACAGCCTGAC 
                 
               
               
                   
               
               
                 
                   CTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCACGAGCACATTGCCA 
                 
               
               
                   
               
               
                 
                   ATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGGTGGACGAGCTCGTG 
                 
               
               
                   
               
               
                 
                   AAAGTGATGGGCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCCA 
                 
               
               
                   
               
               
                 
                   GAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAAAGAGCTGGGCAGCC 
                 
               
               
                   
               
               
                 
                   AGATCCTGAAAGAACACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTG 
                 
               
               
                   
               
               
                 
                   CAGAATGGGCGGGATATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGA 
                 
               
               
                   
               
               
                 
                   CGCCATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCTGACCAGAAGCGACA 
                 
               
               
                   
               
               
                 
                   AGAACCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGAAGAACTACTGGCGG 
                 
               
               
                   
               
               
                 
                   CAGCTGCTGAACGCCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGGCGG 
                 
               
               
                   
               
               
                 
                   CCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCGGCAGATCACAAAGC 
                 
               
               
                   
               
               
                 
                   ACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACAAGCTGATCCGGGAA 
                 
               
               
                   
               
               
                 
                   GTGAAAGTGATCACCCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGT 
                 
               
               
                   
               
               
                 
                   GCGCGAGATCGGCAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGTGGGAACCGCCCTGA 
                 
               
               
                   
               
               
                 
                   TCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGTACGACGTGCGGAAG 
                 
               
               
                   
               
               
                 
                   ATGATCGCCAAGAGCGAGCAGGAAATCGGCAAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCAT 
                 
               
               
                   
               
               
                 
                   GAACTTTTTCAAGACCAGATTACCCTGGCCAACGGCGAGAGTCCGGAAGCGGCCTCTGATCGAGACAA 
                 
               
               
                   
               
               
                 
                   ACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGCGGAAAGTGCTGAGC 
                 
               
               
                   
               
               
                 
                   ATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCT 
                 
               
               
                   
               
               
                 
                   GCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCT 
                 
               
               
                   
               
               
                 
                   TCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAA 
                 
               
               
                   
               
               
                 
                   CTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCAT 
                 
               
               
                   
               
               
                 
                   CGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACT 
                 
               
               
                   
               
               
                 
                   CCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAGAAGGGAAAC 
                 
               
               
                   
               
               
                 
                   GAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTATGAGAAGCTGAAGGG 
                 
               
               
                   
               
               
                 
                   CTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTACCTGGACGAGATCA 
                 
               
               
                   
               
               
                 
                   TCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCC 
                 
               
               
                   
               
               
                 
                   GCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCT 
                 
               
               
                   
               
               
                 
                   GACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCA 
                 
               
               
                   
               
               
                 
                   GCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGATC 
                 
               
               
                   
               
               
                   GACCTGTCTCAGCTGGGAGGCGAC aagcgacctgccgccacaaagaaggctggacaggctaagaagaa 
               
               
                   
               
               
                 gaaactggactctggaggatccgactacaaagaccatgacggtgattataaagatcatgacatcgatt 
               
               
                   
               
               
                 acaaggatgacgatgacaagggaggatccaaggagaagagtgcttgtcctaaagatccagccaaacct 
               
               
                   
               
               
                 ccggccaaggcacaagttgtgggatggccaccggtgagatcataccggaagaacgtgatggtttcctg 
               
               
                   
               
               
                 ccaaaaatcaagcggtggcccggaggcggcggcgttcgtgaaggtatcaatggacggagcaccgtact 
               
               
                   
               
               
                 tgaggaaaatcgatttgaggatgtataaaggcggatctggcggctctggaggatcc gatgctttagac   
               
               
                   
               
               
                 
                   gattttgacttagatatgcttggttcagacgcgttagacgacttcgacctagacatgttaggctcaga 
                 
               
               
                   
               
               
                 
                   tgcattggacgacttcgatttagatatgttgggctccgatgccctagatgactttgatctagatatgc 
                 
               
               
                   
               
               
                 
                   taggtagtcccaaaaagaagaggaaagtgggatcccagtatctgcccgacacagatgatagacaccga 
                 
               
               
                   
               
               
                 
                   atcgaagagaaacgcaagcgaacgtatgaaaccttcaaatcgatcatgaagaaatcgcccttctoggg 
                 
               
               
                   
               
               
                 
                   tccgaccgatcccaggcccccaccgagaaggattgcggtcccgtcccgctcgtcggccagcgtgccga 
                 
               
               
                   
               
               
                 
                   agcctgcgccgcagccctaccccttcacgtcgagcctgagcacaatcaattatgacgagttcccgacg 
                 
               
               
                   
               
               
                 
                   atggtgttcccctcgggacaaatctcacaagcctcggcgctcgcaccagcgcctccccaagtccttcc 
                 
               
               
                   
               
               
                 
                   gcaagcgcctgccccagcgcctgcaccggcaatggtgtccgccctcgcacaggcccctgcgcccgtcc 
                 
               
               
                   
               
               
                 
                   ccgtgctcgcgcctggaccgccccaggcggtcgctccaccggctccgaagccgacgcaggccggagag 
                 
               
               
                   
               
               
                 
                   ggaacactctccgaagcacttcttcaactccagtttgatgacgaggatcttggagcactccttggaaa 
                 
               
               
                   
               
               
                 
                   ctcgacagaccctgcggtgtttaccgacctcgcgtcagtagataactccgaatttcagcagcttttga 
                 
               
               
                   
               
               
                 
                   accagggtatcccggtcgcgccacatacaacggagcccatgttgatggaataccccgaagcaatcacg 
                 
               
               
                   
               
               
                 
                   agacttgtgacgggagcgcagcggcctcccgatcccgcacccgcacctttgggggcacctggcctccc 
                 
               
               
                   
               
               
                 
                   taacggacttttgagcggcgacgaggatttctcctccatcgccgatatggatttctcagccttgctgt 
                 
               
               
                   
               
               
                 
                   cacagatttccagcggctctggcagcggcagccgggattccagggaagggatgtttttgccgaagcct 
                 
               
               
                   
               
               
                 
                   gaggccggctccgctattagtgacgtgtttgagggccgcgaggtgtgccagccaaaacgaatccggcc 
                 
               
               
                   
               
               
                 
                   atttcatcctccaggaagtccatgggccaaccgcccactccccgccagcctcgcaccaacaccaaccg 
                 
               
               
                   
               
               
                 
                   gtccagtacatgagccagtcgggtcactgaccccggcaccagtccctcagccactggatccagcgccc 
                 
               
               
                   
               
               
                 
                   gcagtgactcccgaggccagtcacctattggaggatcccgatgaagagacgagccaggctgtcaaagc 
                 
               
               
                   
               
               
                 
                   ccttcgggagatggccgatactgtgattccccagaaggaagaggctgcaatctgtagccaaatggacc 
                 
               
               
                   
               
               
                 
                   tttcccatccgcccccaaggggccatctggatgagctgacaaccacacttgagtccatgaccgaggat 
                 
               
               
                   
               
               
                 
                   ctgaacctggactcacccctgaccccggaattgaacgagattctggataccttcctgaacgacgagtg 
                 
               
               
                   
               
               
                 
                   cctcttgcatgccatgcatatcagcacaggactgtccatcttcgacacatctctgttttga 
                 
               
               
                   
               
               
                 Amino acid sequence fordCas9-VPR 
               
               
                 (corresponding to SEQ ID NO: 77); lowercase 
               
               
                 underlined = VPR; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 78 
                   
               
               
                 
                   MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                 
                   
               
               
                   
               
               
                 
                   RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIY 
                 
               
               
                   
               
               
                 
                   HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
                 
               
               
                   
               
               
                 
                   GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
                 
               
               
                   
               
               
                 
                   DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
                 
               
               
                   
               
               
                 
                   QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
                 
               
               
                   
               
               
                 
                   SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
                 
               
               
                   
               
               
                 
                   NFEEVVDKGASAQSFTERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQ 
                 
               
               
                   
               
               
                 
                   KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
                 
               
               
                   
               
               
                 
                   EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
                 
               
               
                   
               
               
                 
                   DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
                 
               
               
                   
               
               
                 
                   KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
                 
               
               
                   
               
               
                 
                   QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
                 
               
               
                   
               
               
                 
                   QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
                 
               
               
                   
               
               
                 
                   VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
                 
               
               
                   
               
               
                 
                   MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
                 
               
               
                   
               
               
                 
                   MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKK 
                 
               
               
                   
               
               
                 
                   LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
                 
               
               
                   
               
               
                 
                   ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLS 
                 
               
               
                   
               
               
                 
                   AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
                 
               
               
                   
               
               
                   DLSQLGGD krpaatkkaqqakkkkldsggsdykdhdgdykdhdidykddddkggskeksacpkdpakp 
               
               
                   
               
               
                 pakaqvvgwppvrsyrknvmvscqkssggpeaaafvkvsmdgapylrkidlrmykggsggsggs dald   
               
               
                   
               
               
                 
                   dfdldmlgsdalddfdldmlgsdalddfdldmlgsdalddfdldmlgspkkkrkvgsqylpdtddrhr 
                 
               
               
                   
               
               
                 
                   ieekrkrtyetfksimkkspfsgptdprppprriavpsrssasypkpapqpypftsslstinydefpt 
                 
               
               
                   
               
               
                 
                   mvfpsgqisqasalapappqvlpqapapapapamvsalaqapapvpvlapgppqavappapkptqaqe 
                 
               
               
                   
               
               
                 
                   gtlseallqlqfddedlgallgnstdpavftdlasvdnsefqqllnqgipvaphttepmlmeypeait 
                 
               
               
                   
               
               
                 
                   rlvtgaqrppdpapaplqapglpngllsgdedfssiadmdfsallsqissgsgsgsrdsregmflpkp 
                 
               
               
                   
               
               
                 
                   eagsaisdvfegrevcqpkrirpfhppgspwanrplpaslaptptgpvhepvqsllpapvpqpldpap 
                 
               
               
                   
               
               
                 
                   avtpeashlledpdeetsqavkalremadtvipqkeeaaicgqmdlshppprghldeltttlesmted 
                 
               
               
                   
               
               
                 
                   lnldspllpelneildtflndecllhamhistglsifdtslf* 
                 
               
               
                   
               
               
                 DNA sequence for dCas9-p300c (in backbone pN/36): 
               
               
                 pNI37; lowercase underlined = p300c; 
               
               
                 capital underlined = dCas9, 
               
               
                 SEQ ID NO: 79 
                   
               
               
                 
                   ATGGACAAGAAGTACAGCATCGGCCTGGCCATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGA 
                 
                   
               
               
                   
               
               
                 
                   CGAGTACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAAGAAGA 
                 
               
               
                   
               
               
                 
                   ACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGCTGAAGAGAACCGCC 
                 
               
               
                   
               
               
                 
                   AGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGC 
                 
               
               
                   
               
               
                 
                   CAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAAGCACG 
                 
               
               
                   
               
               
                 
                   AGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTAC 
                 
               
               
                   
               
               
                 
                   CACCTGAGAAAGAAACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGC 
                 
               
               
                   
               
               
                 
                   CCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGG 
                 
               
               
                   
               
               
                 
                   ACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGC 
                 
               
               
                   
               
               
                 
                   GGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAATCTGATCGC 
                 
               
               
                   
               
               
                 
                   CCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAGCCTGGGCCTGACCC 
                 
               
               
                   
               
               
                 
                   CCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCAAGGACACCTACGAC 
                 
               
               
                   
               
               
                 
                   GACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTTCTGGCCGCCAAGAA 
                 
               
               
                   
               
               
                 
                   CCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGA 
                 
               
               
                   
               
               
                 
                   GCGCCTCTATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGTGCGG 
                 
               
               
                   
               
               
                 
                   CAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGCTACGCCGGCTACAT 
                 
               
               
                   
               
               
                 
                   TGACGGCGGAGCCAGCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCA 
                 
               
               
                   
               
               
                 
                   CCGAGGAACTGCTCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAACGGC 
                 
               
               
                   
               
               
                 
                   AGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAGGAAGATTTTTACCC 
                 
               
               
                   
               
               
                 
                   ATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCCCTACTACGTGGGCC 
                 
               
               
                   
               
               
                 
                   CTCTGGCCAGGGGAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGG 
                 
               
               
                   
               
               
                 
                   AACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGGATGACCAACTTCGA 
                 
               
               
                   
               
               
                 
                   TAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTACTTCACCGTGTATA 
                 
               
               
                   
               
               
                 
                   ACGAGCTGACCAAAGTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAG 
                 
               
               
                   
               
               
                 
                   AAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCAGCTGAAAGAGGA 
                 
               
               
                   
               
               
                 
                   CTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCCGGCGTGGAAGATCGGTTCAACGCCT 
                 
               
               
                   
               
               
                 
                   CCCTGGGCACATACCACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAAC 
                 
               
               
                   
               
               
                 GAGGACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCGAGGAACG 
               
               
                   
               
               
                 
                   GCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCGGCGGAGATACACCG 
                 
               
               
                   
               
               
                 
                   GCTGGGGCAGGCTGAGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTG 
                 
               
               
                   
               
               
                 
                   GATTTCCTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACGACAGCCTGAC 
                 
               
               
                   
               
               
                 
                   CTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCACGAGCACATTGCCA 
                 
               
               
                   
               
               
                 
                   ATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGGTGGACGAGCTCGTG 
                 
               
               
                   
               
               
                 
                   AAAGTGATGGGCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCCA 
                 
               
               
                   
               
               
                 
                   GAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAAAGAGCTGGGCAGCC 
                 
               
               
                   
               
               
                 
                   AGATCCTGAAAGAACACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTG 
                 
               
               
                   
               
               
                 
                   CAGAATGGGCGGGATATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGA 
                 
               
               
                   
               
               
                 
                   CGCCATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCTGAGGAGAAGCGACA 
                 
               
               
                   
               
               
                 
                   AGAACCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGAAGAACTACTGGCGG 
                 
               
               
                   
               
               
                 
                   CAGCTGCTGAACGCCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGGCGG 
                 
               
               
                   
               
               
                 
                   CCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCGGCAGATCACAAAGC 
                 
               
               
                   
               
               
                 
                   ACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACAAGCTGATCCGGGAA 
                 
               
               
                   
               
               
                 
                   GTGAAAGTGATCACCCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGT 
                 
               
               
                   
               
               
                 
                   GCGCGAGATCAACAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGTGGGAACCGCCCTGA 
                 
               
               
                   
               
               
                 
                   TCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGTACGACGTGCGGAAG 
                 
               
               
                   
               
               
                 
                   ATGATCGCCAAGAGCGAGCAGGAAATCGGCAAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCAT 
                 
               
               
                   
               
               
                 
                   GAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCCTCTGATCGAGACAA 
                 
               
               
                   
               
               
                 
                   ACGGCGAAACCGGGGAGATCGTGGTGGGATAAGGGCCGGGATTTTTGCCACCGTGCGAAAGTGTGAGC 
                 
               
               
                   
               
               
                 
                   ATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCT 
                 
               
               
                   
               
               
                 
                   GCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCT 
                 
               
               
                   
               
               
                 
                   TCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAA 
                 
               
               
                   
               
               
                 
                   CTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCAT 
                 
               
               
                   
               
               
                 
                   CGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACT 
                 
               
               
                   
               
               
                 
                   CCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAGAAGGGAAAC 
                 
               
               
                   
               
               
                 
                   GAACTGGCCCTGCCCTuCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTATGAGAAGCTGAAGGG 
                 
               
               
                   
               
               
                 
                   CTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTACCTGGACGAGATCA 
                 
               
               
                   
               
               
                 
                   TCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCC 
                 
               
               
                   
               
               
                 
                   GCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCT 
                 
               
               
                   
               
               
                 
                   GACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCA 
                 
               
               
                   
               
               
                 
                   GCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGATC 
                 
               
               
                   
               
               
                   GACCTGTCTCAGCTGGGAGGCGAC aagcgacctgccgccacaaagaaggctggacaggctaagaagaa 
               
               
                   
               
               
                 gaaactggactctggaggatccgactacaaagaccatgacggtgattataaagatcatgacatcgatt 
               
               
                   
               
               
                 acaaggatgacgatgacaagggaggatccaaggagaagagtgcttgtcctaaagatccagccaaacct 
               
               
                   
               
               
                 ccggccaaggcacaagttgtgggatggccaccggtgagatcataccggaagaacgtgatggtttcctg 
               
               
                   
               
               
                 ccaaaaatcaagcggtggcccggaggcggcggcgttcgtgaaggtatcaatggacggagcaccgtact 
               
               
                   
               
               
                 tgaggaaaatcgatttgaggatgtataaaggcggatctggcggctctggaggatccattttcaaacca 
               
               
                   
               
               
                 
                   gaagaactacgacaggcactgatgccaactttggaggcactttaccgtcaggatccagaatcccttcc 
                 
               
               
                   
               
               
                 
                   ctttcgtcaacctgtggaccctcagcttttaggaatccctgattactttgatattgtgaagagcccca 
                 
               
               
                   
               
               
                 
                   tggatctttctaccattaagaggaagttagacactggacagtatcaggagccctggcagtatgtcgat 
                 
               
               
                   
               
               
                 
                   gatatttggcttatgttcaataatgcctggttatataaccggaaaacatcacgggtatacaaatactg 
                 
               
               
                   
               
               
                 
                   ctccaagctctctgaggtctttgaacaagaaattgacccagtgatgcaaagccttggatactgttgtg 
                 
               
               
                   
               
               
                 
                   gcagaaagttggagttctctccacagacactgtgttgctacggcaaacagttgtgcacaatacctcgt 
                 
               
               
                   
               
               
                 
                   gatgccacttattacagttaccagaacaggtatcatttctgtgagaagtgtttcaatgagatccaagg 
                 
               
               
                   
               
               
                 
                   ggagagcgtttctttgggggatgacccttcccagcctcaaactacaataaataaagaacaattttcca 
                 
               
               
                   
               
               
                 
                   agagaaaaaatgacacactggatcctgaactgtttgttgaatgtacagagtgcggaagaaagatgcat 
                 
               
               
                   
               
               
                 
                   cagatctgtatccttcaccatgagatcatctggcctgctggattcgtctgtgatggctgtttaaagaa 
                 
               
               
                   
               
               
                 
                   aagtgcacgaactaggaaagaaaataagttttctgctaaaaggttgccatctaccagacttggcacct 
                 
               
               
                   
               
               
                 
                   ttctagagaatcgtgtgaatgactttctgaggcgacagaatcaccctgagtcaggagaggtcactgtt 
                 
               
               
                   
               
               
                 
                   agagtagttcatgcttctgacaaaaccgtggaagtaaaaccaggcatgaaagcaaggtttgtggacag 
                 
               
               
                   
               
               
                 
                   tggagagatggcagaatcctttccataccgaaccaaagccctctttgcctttgaagaaattgatggtg 
                 
               
               
                   
               
               
                 
                   ttgacctgtgcttctttggcatgcatgttcaagagtatggctctgactgccctccacccaaccagagg 
                 
               
               
                   
               
               
                 
                   agagtatacatatcttacctcgatagtgttcatttcttccgtcctaaatgcttgaggactgcagtcta 
                 
               
               
                   
               
               
                 
                   tcatgaaatcctaattggatatttagaatatgtcaagaaattaggttacacaacagggcatatttggg 
                 
               
               
                   
               
               
                 
                   catgtccaccaagtgagggagatgattatatcttccattgccatcctcctgaccagaagatacccaag 
                 
               
               
                   
               
               
                 
                   cccaagcgactgcaggaatggtacaaaaaaatgcttgacaaggctgtatcagagcgtattgtccatga 
                 
               
               
                   
               
               
                 
                   ctacaaggatatttttaaacaagctactgaagatagattaacaagtgcaaaggaattgccttatttcg 
                 
               
               
                   
               
               
                 
                   agggtgatttctggcccaatgttctggaagaaagcattaaggaactggaacaggaggaagaagagaga 
                 
               
               
                   
               
               
                 
                   aaacgagaggaaaacaccagcaatgaaagcacagatgtgaccaagggagacagcaaaaatgctaaaaa 
                 
               
               
                   
               
               
                 
                   gaagaataataagaaaaccagcaaaaataagagcagcctgagtaggggcaacaagaagaaacccggga 
                 
               
               
                   
               
               
                 
                   tgcccaatgtatctaacgacctctcacagaaactatatgccaccatggagaagcataaagaggtcttc 
                 
               
               
                   
               
               
                 
                   tttgtgatccgcctcattgctggccctgctgccaactccctgcctcccattgttgatcctgatcctct 
                 
               
               
                   
               
               
                 
                   catcccctgcgatctgatggatggtcgggatgcgtttctcacgctggcaagggacaagcacctggagt 
                 
               
               
                   
               
               
                 
                   tctcttcactccgaagagcccagtggtccaccatgtgcatgctggtggagctgcacacgcagagccag 
                 
               
               
                   
               
               
                 
                   gactga 
                 
               
               
                   
               
               
                 Amino acid sequence fordCas9-p300c 
               
               
                 (corresponding to SEQ ID NO: 79); lowercase 
               
               
                 underlined = p300c; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 80 
                   
               
               
                 
                   MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                 
                   
               
               
                   
               
               
                 
                   RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTTY 
                 
               
               
                   
               
               
                 
                   HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
                 
               
               
                   
               
               
                 
                   GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
                 
               
               
                   
               
               
                 
                   DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
                 
               
               
                   
               
               
                 
                   QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
                 
               
               
                   
               
               
                 
                   SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
                 
               
               
                   
               
               
                 
                   NFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQ 
                 
               
               
                   
               
               
                 
                   KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
                 
               
               
                   
               
               
                 
                   EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
                 
               
               
                   
               
               
                 
                   DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
                 
               
               
                   
               
               
                 
                   KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKKLEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
                 
               
               
                   
               
               
                 
                   QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
                 
               
               
                   
               
               
                 
                   QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
                 
               
               
                   
               
               
                 
                   VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
                 
               
               
                   
               
               
                 
                   MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
                 
               
               
                   
               
               
                 
                   MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKK 
                 
               
               
                   
               
               
                 
                   LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
                 
               
               
                   
               
               
                 
                   ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLS 
                 
               
               
                   
               
               
                 
                   AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
                 
               
               
                   
               
               
                   DLSQLGGD krpaatkkaqqakkkkldsggsdykdhdgdykdhdidykddddkggskeksacpkdpakp 
               
               
                   
               
               
                 pakaqvvgwppvrsyrknvmvscqkssggpeaaafvkvsmdgapylrkidlrmykggsggsggs ifkp   
               
               
                   
               
               
                 
                   eelrqalmptlealyrqdpeslprfqpvapqllgipdyfdivkspmdlstlkrkldtgqyqapwqyvd 
                 
               
               
                   
               
               
                 
                   diwlmfnnawlynrktsrvykycsklsevfeqeidpvmgslgyccgrklefspqtlccyqkqlctipr 
                 
               
               
                   
               
               
                 
                   datyysyqnryhfcekcfneiqgesvslgddpsqpqttinkeqfskrkndtldpelfvectecgrkmh 
                 
               
               
                   
               
               
                 
                   qicvlhheiiwpagfvcdgclkksartrkenkfsakrlpstrlgtflenrvndfIrrqnhpesgevtv 
                 
               
               
                   
               
               
                 
                   rvvhasdktvevkpgmkarfvdsgemaesfpyrtkalfafeeidgvdlcffgmhvqeygsdcpppngr 
                 
               
               
                   
               
               
                 
                   rvyisyldsvhffrpkclrtavyheiligyleyvkklgyttghiwacppsegddyifhchppdqkipk 
                 
               
               
                   
               
               
                 
                   pkrlqewykkmldkavserivhdykdifkqatedrltsakeipyfeqdfwpnvleesikelegeeeer 
                 
               
               
                   
               
               
                 
                   kreentsnestdvtkgdsknakkknnkktsknksslsrgnkkkpgmpnvsndlsqklyatmekhkevf 
                 
               
               
                   
               
               
                 
                   fvirliagpaanslppivdpdplipcdlmdgrdafltlardkhlefsslrraqwstmcmlvelhtqsq 
                 
               
               
                   
               
               
                 
                   d* 
                 
               
               
                   
               
               
                 DNA sequence for dCas9-p300c (in backbone pNI123): pNH33; lowercase 
               
               
                 underlined = p300c; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 81 
                   
               
               
                 
                   ATGGACAAGAAGTACTCCATTGGGCTCGCCATCGGCACAAACAGCGTCGGCTGGGCCGTCATTACGGA 
                 
                   
               
               
                   
               
               
                 
                   CGAGTACAAGGTGCCGAGCAAAAAATTCAAAGTTCTGGGCAATACCGATCGCCACAGCATAAAGAAGA 
                 
               
               
                   
               
               
                 
                   ACCTCATTGGCGCCCTCCTGTTCGACTCCGGGGAAACCGCCGAAGCCACGCGGCTCAAAAGAACAGCA 
                 
               
               
                   
               
               
                 
                   CGGCGCAGATATACCCGCAGAAAGAATCGGATCTGCTACCTGCAGGAGATCTTTAGTAATGAGATGGC 
                 
               
               
                   
               
               
                 
                   TAAGGTGGATGACTCTTTCTTCCATAGGCTGGAGGAGTCCTTTTTGGTGGAGGAGGATAAAAAGCACG 
                 
               
               
                   
               
               
                 
                   AGCGCCACCCAATCTTTGGCAATATCGTGGACGAGGTGGCGTACCATGAAAAGTACCCAACCATATAT 
                 
               
               
                   
               
               
                 
                   CATCTGAGGAAGAAGCTTGTAGACAGTACTGATAAGGCTGACTTGCGGTTGATCTATCTCGCGCTGGC 
                 
               
               
                   
               
               
                 
                   GCATATGATCAAATTTCGGGGACACTTCCTCATCGAGGGGGACCTGAACCCAGACAACAGCGATGTCG 
                 
               
               
                   
               
               
                 
                   ACAAACTCTTTATCCAACTGGTTCAGACTTACAATCAGCTTTTCGAAGAGAACCCGATCAACGCATCC 
                 
               
               
                   
               
               
                 
                   GGAGTTGACGCCAAAGCAATCCTGAGCGCTAGGCTGTCCAAATCCCGGCGGCTCGAAAACCTCATCGC 
                 
               
               
                   
               
               
                 
                   ACAGCTCCCTGGGGAGAAGAAGAACGGCCTGTTTGGTAATCTTATCGCCCTGTCACTCGGGCTGACCC 
                 
               
               
                   
               
               
                 
                   CCAACTTTAAATCTAACTTCGACCTGGCCGAAGATGCCAAGCTTCAACTGAGCAAAGACACCTACGAT 
                 
               
               
                   
               
               
                 
                   GATGATCTCGACAATCTGCTGGCCCAGATCGGCGACCAGTACGCAGACCTTTTTTTGGCGGCAAAGAA 
                 
               
               
                   
               
               
                 
                   CCTGTCAGACGCCATTCTGCTGAGTGATATTCTGCGAGTGAACACGGAGATCACCAAAGCTCCGCTGA 
                 
               
               
                   
               
               
                 
                   GCGCTAGTATGATCAAGCGCTATGATGAGCACCACCAAGACTTGACTTTGCTGAAGGCCCTTGTCAGA 
                 
               
               
                   
               
               
                 
                   CAGCAACTGCCTGAGAAGTACAAGGAAATTTTCTTCGATCAGTCTAAAAATGGCTACGCCGGATACAT 
                 
               
               
                   
               
               
                 
                   TGACGGCGGAGCAAGCCAGGAGGAATTTTACAAATTTATTAAGCCCATCTTGGAAAAAATGGACGGCA 
                 
               
               
                   
               
               
                 
                   CCGAGGAGCTGCTGGTAAAGCTTAACAGAGAAGATCTGTTGCGCAAACAGCGCACTTTCGACAATGGA 
                 
               
               
                   
               
               
                 
                   AGCATCCCCCACCAGATTCACCTGGGCGAACTGCACGCTATCCTCAGGCGGCAAGAGGATTTCTACCC 
                 
               
               
                   
               
               
                 
                   CTTTTTGAAAGATAACAGGGAAAAGATTGAGAAAATCCTCACATTTCGGATACCCTACTATGTAGGCC 
                 
               
               
                   
               
               
                 
                   CCCTCGCCCGGGGAAATTCCAGATTCGCGTGGATGACTCGCAAATCAGAAGAGACCATCACTCCCTGG 
                 
               
               
                   
               
               
                 
                   AACTTCGAGGAAGTCGTGGATAAGGGGGCCTCTGCCCAGTCCTTCATCGAAAGGATGACTAACTTTGA 
                 
               
               
                   
               
               
                 
                   TAAAAATCTGCCTAACGAAAAGGTGCTTCCTAAACACTCTCTGCTGTACGAGTACTTCACAGTTTATA 
                 
               
               
                   
               
               
                 
                   ACGAGCTCACCAAGGTCAAATACGTCACAGAAGGGATGAGAAAGCCAGCATTCCTGTCTGGAGAGCAG 
                 
               
               
                   
               
               
                 
                   AAGAAAGCTATCGTGGACCTCCTCTTCAAGACGAACCGGAAAGTTACCGTGAAACAGCTCAAAGAAGA 
                 
               
               
                   
               
               
                 
                   CTATTTCAAAAAGATTGAATGTTTCGACTCTGTTGAAATCAGCGGAGTGGAGGATCGCTTCAACGCAT 
                 
               
               
                   
               
               
                 
                   CCCTGGGAACGTATCACGATCTCCTGAAAATCATTAAAGACAAGGACTTCCTGGACAATGAGGAGAAC 
                 
               
               
                   
               
               
                 
                   GAGGACATTCTTGAGGACATTGTCCTCACCCTTACGTTGTTTGAAGATAGGGAGATGATTGAAGAACG 
                 
               
               
                   
               
               
                 
                   CTTGAAAACTTACGCTCATCTCTTCGACGACAAAGTCATGAAACAGCTCAAGAGGCGCCGATATACAG 
                 
               
               
                   
               
               
                 
                   GATGGGGGCGGCTGTCAAGAAAACTGATCAATGGGATCCGAGACAAGCAGAGTGGAAAGACAATCCTG 
                 
               
               
                   
               
               
                 
                   GATTTTCTTAAGTCCGATGGATTTGCCAACCGGAACTTCATGCAGTTGATCCATGATGACTCTCTCAC 
                 
               
               
                   
               
               
                 
                   CTTTAAGGAGGACATCCAGAAAGCACAAGTTTCTGGCCAGGGGGACAGTCTTCACGAGCACATCGCTA 
                 
               
               
                   
               
               
                 
                   ATCTTGCAGGTAGCCCAGCTATCAAAAAGGGAATACTGCAGACCGTTAAGGTCGTGGATGAACTCGTC 
                 
               
               
                   
               
               
                 
                   AAAGTAATGGGAAGGCATAAGCCCGAGAATATCGTTATCGAGATGGCCCGAGAGAACCAAACTACCCA 
                 
               
               
                   
               
               
                 
                   GAAGGGACAGAAGAACAGTAGGGAAAGGATGAAGAGGATTGAAGAGGGTATAAAAGAACTGGGGTCCC 
                 
               
               
                   
               
               
                 
                   AAATCCTTAAGGAACACCCAGTTGAAAACACCCAGCTTCAGAATGAGAAGCTCTACCTGTACTACCTG 
                 
               
               
                   
               
               
                 
                   CAGAACGGCAGGGACATGTACGTGGATCAGGAACTGGACATCAATCGGCTCTCCGACTACGACGTGGA 
                 
               
               
                   
               
               
                 
                   TGCCATCGTGCCCCAGTCTTTTCTCAAAGATGATTCTATTGATAATAAAGTGTTGACAAGATCCGATA 
                 
               
               
                   
               
               
                 
                   AAAATAGAGGGAAGAGTGATAACGTCCCCTCAGAAGAAGTTGTCAAGAAAATGAAAAATTATTGGCGG 
                 
               
               
                   
               
               
                 
                   CAGCTGCTGAACGCCAAACTGATCACACAACGGAAGTTCGATAATCTGACTAAGGCTGAACGAGGTGG 
                 
               
               
                   
               
               
                 
                   CCTGTCTGAGTTGGATAAAGCCGGCTTCATCAAAAGGCAGCTTGTTGAGACACGCCAGATCACCAAGC 
                 
               
               
                   
               
               
                 
                   ACGTGGCCCAAATTCTCGATTCACGCATGAACACCAAGTACGATGAAAATGACAAACTGATTCGAGAG 
                 
               
               
                   
               
               
                 
                   GTGAAAGTTATTACTCTGAAGTCTAAGCTGGTCTCAGATTTCAGAAAGGACTTTCAGTTTTATAAGGT 
                 
               
               
                   
               
               
                 
                   GAGAGAGATCAACAATTACCACCATGCGCATGATGCCTACCTGAATGCAGTGGTAGGCACTGCACTTA 
                 
               
               
                   
               
               
                 
                   TCAAAAAATATCCCAAGCTTGAATCTGAATTTGTTTACGGAGACTATAAAGTGTACGATGTTAGGAAA 
                 
               
               
                   
               
               
                 
                   ATGATCGCAAAGTCTGAGCAGGAAATAGGCAAGGCCACCGCTAAGTACTTCTTTTACAGCAATATTAT 
                 
               
               
                   
               
               
                 
                   GAATTTTTTCAAGACCGAGATTACACTGGCCAATGGAGAGATTCGGAAGCGACCACTTATCGAAACAA 
                 
               
               
                   
               
               
                 
                   ACGGAGAAACAGGAGAAATCGTGTGGGACAAGGGTAGGGATTTCGCGACAGTCCGGAAGGTCCTGTCC 
                 
               
               
                   
               
               
                 
                   ATGCCGCAGGTGAACATCGTTAAAAAGACCGAAGTACAGACCGGAGGCTTCTCCAAGGAAAGTATCCT 
                 
               
               
                   
               
               
                 
                   CCCGAAAAGGAACAGCGACAAGCTGATCGCACGCAAAAAAGATTGGGACCCCAAGAAATACGGCGGAT 
                 
               
               
                   
               
               
                 
                   TCGATTCTCCTACAGTCGCTTACAGTGTACTGGTTGTGGCCAAAGTGGAGAAAGGGAAGTCTAAAAAA 
                 
               
               
                   
               
               
                 
                   CTCAAAAGCGTCAAGGAACTGCTGGGCATCACAATCATGGAGCGATCAAGCTTCGAAAAAAACCCCAT 
                 
               
               
                   
               
               
                 
                   CGACTTTCTCGAGGCGAAAGGATATAAAGAGGTCAAAAAAGACCTCATCATTAAGCTTCCCAAGTACT 
                 
               
               
                   
               
               
                 
                   CTCTCTTTGAGCTTGAAAACGGCCGGAAACGAATGCTCGCTAGTGCGGGCGAGCTGCAGAAAGGTAAC 
                 
               
               
                   
               
               
                 
                   GAGCTGGCACTGCCCTCTAAATACGTTAATTTCTTGTATCTGGCCAGCCACTATGAAAAGCTCAAAGG 
                 
               
               
                   
               
               
                 
                   GTCTCCCGAAGATAATGAGCAGAAGCAGCTGTTCGTGGAACAACACAAACACTACCTTGATGAGATCA 
                 
               
               
                   
               
               
                 
                   TCGAGCAAATAAGCGAATTCTCCAAAAGAGTGATCCTCGCCGACGCTAACCTCGATAAGGTGCTTTCT 
                 
               
               
                   
               
               
                 
                   GCTTACAATAAGCACAGGGATAAGCCCATCAGGGAGCAGGCAGAAAACATTATCCACTTGTTTACTCT 
                 
               
               
                   
               
               
                 
                   GACCAACTTGGGCGCGCCTGCAGCCTTCAAGTACTTCGACACCACCATAGACAGAAAGCGGTACACCT 
                 
               
               
                   
               
               
                 
                   CTACAAAGGAGGTCCTGGACGCCACACTGATTCATCAGTCAATTACGGGGCTCTATGAAACAAGAATC 
                 
               
               
                   
               
               
                   GACCTCTCTCAGCTCGGTGGAGAC agcagggctgaccccaagaagaagaggaaggtggctagc atttt   
               
               
                   
               
               
                 
                   caaaccagaagaactacgacaggcactgatgccaactttggaggcactttaccgtcaggatccagaat 
                 
               
               
                   
               
               
                 
                   cccttccctttcgtcaacctgtggaccctcagcttttaggaatccctgattactttgatattgtgaag 
                 
               
               
                   
               
               
                 
                   agccccatggatctttctaccattaagaggaagttagacactggacagtatcaggagccctggcagta 
                 
               
               
                   
               
               
                 
                   tgtcgatgatatttggcttatgttcaataatgcctggttatataaccggaaaacatcacgggtataca 
                 
               
               
                   
               
               
                 
                   aatactgctccaagctctctgaggtctttgaacaagaaattgacccagtgatgcaaagccttggatac 
                 
               
               
                   
               
               
                 
                   tgttgtggcagaaagttggagttctctccacagacactgtgttgctacggcaaacagttgtgcacaat 
                 
               
               
                   
               
               
                 
                   acctcgtgatgccacttattacagttaccagaacaggtatcatttctgtgagaagtgtttcaatgaga 
                 
               
               
                   
               
               
                 
                   tccaaggggagagcgtttctttgggggatgacccttcccagcctcaaactacaataaataaagaacaa 
                 
               
               
                   
               
               
                 
                   ttttccaagagaaaaaatgacacactggatcctgaactgtttgttgaatgtacagagtgcggaagaaa 
                 
               
               
                   
               
               
                 
                   gatgcatcagatctgtgtccttcaccatgagatcatctggcctgctggattcgtctgtgatggctgtt 
                 
               
               
                   
               
               
                 
                   taaagaaaagtgcacgaactaggaaagaaaataagttttctgctaaaaggttgccatctaccagactt 
                 
               
               
                   
               
               
                 
                   ggcacctttctagagaatcgtgtgaatgactttctgaggcgacagaatcaccctgagtcaggagaggt 
                 
               
               
                   
               
               
                 
                   cactgttagagtagttcatgcttctgacaaaaccgtggaagtaaaaccaggcatgaaagcaaggtttg 
                 
               
               
                   
               
               
                 
                   tggacagtggagagatggcagaatcctttccataccgaaccaaagccctctttgcctttgaagaaatt 
                 
               
               
                   
               
               
                 
                   gatggtgttgacctgtgcttctttggcatgcatgttcaagagtatggctctgactgccctccacccaa 
                 
               
               
                   
               
               
                 
                   ccagaggagagtatacatatcttacctcgatagtgttcatttcttccgtcctaaatgcttgaggactg 
                 
               
               
                   
               
               
                 
                   cagtctatcatgaaatcctaattggatatttagaatatgtcaagaaattaggttacacaacagggcat 
                 
               
               
                   
               
               
                 
                   atttgggcatgtccaccaagtgagggagatgattatatcttccattgccatcctcctgaccagaagat 
                 
               
               
                   
               
               
                 
                   acccaagcccaagcgactgcaggaatggtacaaaaaaatgcttgacaaggctgtatcagagcgtattg 
                 
               
               
                   
               
               
                 
                   tccatgactacaaggatatttttaaacaagctactgaagatagattaacaagtgcaaaggaattgcct 
                 
               
               
                   
               
               
                 
                   tatttcgagggtgatttctggcccaatgttctggaagaaagcattaaggaactggaacaggaggaaga 
                 
               
               
                   
               
               
                 
                   agagagaaaacgagaggaaaacaccagcaatgaaagcacagatgtgaccaagggagacagcaaaaatg 
                 
               
               
                   
               
               
                 
                   ctaaaaagaagaataataagaaaaccagcaaaaataagagcagcctgagtaggggcaacaagaagaaa 
                 
               
               
                   
               
               
                 
                   cccggatgcccaatgtatctaacgacctctccacagaaactatatgccaccatggagaagcataaaga 
                 
               
               
                   
               
               
                 
                   ggtcttctttgtgatccgcctcattgctggccctgctgccaactccctgcctcccattgttgatcctg 
                 
               
               
                   
               
               
                 
                   atcctctcatcccctgcgatctgatggatggtcgggatgcgtttctcacgctggcaagggacaagcac 
                 
               
               
                   
               
               
                 
                   ctggagttctcttcactccgaagagcccagtggtccaccatgtgcatgctggtggagctgcacacgca 
                 
               
               
                   
               
               
                 
                   gagccaggac 
                 
               
               
                   
               
               
                 Amino acid sequence fordCas9-p300c 
               
               
                 (corresponding to SEQ ID NO: 81); lowercase 
               
               
                 underlined = p300c; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 82 
                   
               
               
                 MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                   
               
               
                   
               
               
                 
                   RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTTY 
                 
               
               
                   
               
               
                 
                   HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
                 
               
               
                   
               
               
                 
                   GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
                 
               
               
                   
               
               
                 
                   DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
                 
               
               
                   
               
               
                 
                   QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
                 
               
               
                   
               
               
                 
                   SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
                 
               
               
                   
               
               
                 
                   NFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQ 
                 
               
               
                   
               
               
                 
                   KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
                 
               
               
                   
               
               
                 
                   EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
                 
               
               
                   
               
               
                 
                   DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
                 
               
               
                   
               
               
                 
                   KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
                 
               
               
                   
               
               
                 
                   QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
                 
               
               
                   
               
               
                 
                   QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
                 
               
               
                   
               
               
                 
                   VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
                 
               
               
                   
               
               
                 
                   MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
                 
               
               
                   
               
               
                 
                   MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKK 
                 
               
               
                   
               
               
                 
                   LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
                 
               
               
                   
               
               
                 
                   ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLS 
                 
               
               
                   
               
               
                 
                   AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHOSITGLYETRI 
                 
               
               
                   
               
               
                   DLSQLGGDSRAD pkkkrkvas ifkpeelrqalmptlealyrqdpeslpfrqpvdpqllgipdyfdivk   
               
               
                   
               
               
                 
                   spmdlstikrkldtgqyqepwayvddiwlmfnnawlynrktsrvykycsklsevfeceidpvmgslqy 
                 
               
               
                   
               
               
                 
                   ccgrklefspqtlccygkqlctiprdatyysyqnryhfcekcfneiqgesvsIgddpsqpqttinkeq 
                 
               
               
                   
               
               
                 
                   fskrkndtldpelfvectecgrkmhqicvlhheiiwpagfvcdgclkksartrkenkfsakrlpstrl 
                 
               
               
                   
               
               
                 
                   gtflenrvndfirrgnhpesgevtvrvvhasdktvevkpgmkarfvdsgemaesfpyrtkalfafeei 
                 
               
               
                   
               
               
                 
                   davdlcffqmhvqeygsdcpppngrrvyisyldsvhffrpkclrtavyheiligyleyvkklgyttch 
                 
               
               
                   
               
               
                 
                   iwacppsegddyifhchppdqkipkpkrlqewykkmldkavserivhdykdifkqatedrltsakelp 
                 
               
               
                   
               
               
                 
                   yfegdfwpnvleesikeleqeeeerkreentsnestdvtkgdsknakkknnkktsknksslsrgnkkk 
                 
               
               
                   
               
               
                 
                   pgmpnvsndisqklyatmekhkevffvirliagpaanslppivdpdplipcdlmdgrdafltlardkh 
                 
               
               
                   
               
               
                 
                   lefsslrraqwstmcmlvelhtqsqd 
                 
               
               
                   
               
               
                 DNA sequence for P300c-dCas9 (in backbone pNI36): 
               
               
                 pN197; lowercase underlined = p300c; 
               
               
                 capital underlined = dCas9, 
               
               
                 SEQ ID NO: 83 
                   
               
               
                 
                   atgggtattttcaaaccagaagaactacgacaggcactgatgccaactttggaggcactttaccgtca 
                 
                   
               
               
                   
               
               
                 
                   ggatccagaatcccttccctttcgtcaacctgtggaccctcagcttttaggaatccctgattactttg 
                 
               
               
                   
               
               
                 
                   atattgtgaagagccccatggatctttctaccattaagaggaagttagacactggacagtatcaggag 
                 
               
               
                   
               
               
                 
                   ccctggcagtatgtcgatgatatttggcttatgttcaataatgcctggttatataaccggaaaacatc 
                 
               
               
                   
               
               
                 
                   acgggtatacaaatactgctccaagctctctgaggtctttgaacaagaaattgacccagtgatgcaaa 
                 
               
               
                   
               
               
                 
                   gccttggatactgttgtggcagaaagttggagttctctccacagacactgtgttgctacggcaaacag 
                 
               
               
                   
               
               
                 
                   ttgtgcacaatacctcgtgatgccacttattacagttaccagaacaggtatcatttctgtgagaagtg 
                 
               
               
                   
               
               
                 
                   tttcaatgagatccaaggggagagcgtttctttgggggatgacccttcccagcctcaaactacaataa 
                 
               
               
                   
               
               
                 
                   ataaagaacaattttccaagagaaaaaatgacacactggatcctgaactgtttgttgaatgtacagag 
                 
               
               
                   
               
               
                 
                   tgcggaagaaagatgcatcagatctgtgtccttcaccatgagatcatctggcctgctggattcgtctg 
                 
               
               
                   
               
               
                 
                   tgatggctgtttaaagaaaagtgcacgaactaggaaagaaaataagttttctgctaaaaggttgccat 
                 
               
               
                   
               
               
                 
                   ctaccagacttggcacctttctagagaatcgtgtgaatgactttctgaggcgacagaatcaccctgag 
                 
               
               
                   
               
               
                 
                   tcaggagaggtcactgttagagtagttcatgcttctgacaaaaccgtggaagtaaaaccaggcatgaa 
                 
               
               
                   
               
               
                 
                   agcaaggtttgtggacagtggagagatggcagaatcctttccataccgaaccaaagccctctttgcct 
                 
               
               
                   
               
               
                 
                   ttgaagaaattgatggtgttgacctgtgcttctttggcatgcatgttcaagagtatggctctgactgc 
                 
               
               
                   
               
               
                 
                   cctccacccaaccagaggagagtatacatatcttacctcgatagtgttcatttcttccgtcctaaatg 
                 
               
               
                   
               
               
                 
                   cttgaggactgcagtctatcatgaaatcctaattggatatttagaatatgtcaagaaattaggttaca 
                 
               
               
                   
               
               
                 
                   caacagggcatatttgggcatgtccaccaagtgagggagatgattatatcttccattgccatcctcct 
                 
               
               
                   
               
               
                 
                   gaccagaagatacccaagcccaagcgactgcaggaatggtacaaaaaaatgcttgacaaggctgtatc 
                 
               
               
                   
               
               
                 
                   agagcgtattgtccatgactacaaggatatttttaaacaagctactgaagatagattaacaagtgcaa 
                 
               
               
                   
               
               
                 
                   aggaattgccttatttcgagggtgatttctggcccaatgttctggaagaaagcattaaggaactggaa 
                 
               
               
                   
               
               
                 
                   caggaggaagaagagagaaaacgagaggaaaacaccagcaatgaaagcacagatgtgaccaagggaga 
                 
               
               
                   
               
               
                 
                   cagcaaaaatgctaaaaagaagaataataagaaaaccagcaaaaataagagcagcctgagtaggggca 
                 
               
               
                   
               
               
                 
                   acaagaagaaacccgggatgcccaatgtatctaacgacctctcacagaaactatatgccaccatggag 
                 
               
               
                   
               
               
                 
                   aagcataaagaggtcttctttgtgatccgcctcattgctggccctgctgccaactccctgcctcccat 
                 
               
               
                   
               
               
                 
                   tgttgatcctgatcctctcatcccctgcgatctgatggatggtcgggatgcgtttctcacgctggcaa 
                 
               
               
                   
               
               
                 
                   gggacaagcacctggagttctcttcactccgaagagcccagtggtccaccatgtgcatgctggtggag 
                 
               
               
                   
               
               
                   ctgcacacgcagagccaggac ggctctggaggatctggcggctctagcgccacc ATGGACAAGAAGTA   
               
               
                   
               
               
                 
                   CAGCATCGGCCTGGCCATCGGCACCAACTCTGTGGGCTGGGUCGTGATCACCGACGAGTACAAGGTGC 
                 
               
               
                   
               
               
                 
                   CCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAAGAAGAACCTGATCGGAGCC 
                 
               
               
                   
               
               
                 
                   CTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGCTGAAGAGAACCGCCAGAAGAAGATACAC 
                 
               
               
                   
               
               
                 
                   CAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGCCAAGGTGGACGACA 
                 
               
               
                   
               
               
                 
                   GCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAAGCACGAGCGGCACCCCATC 
                 
               
               
                   
               
               
                 
                   TTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTGAGAAAGAA 
                 
               
               
                   
               
               
                 
                   ACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGCCCACATGATCAAGT 
                 
               
               
                   
               
               
                 
                   TCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGGACAAGCTGTTCATC 
                 
               
               
                   
               
               
                 
                   CAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGCGGCGTGGACGCCAA 
                 
               
               
                   
               
               
                 
                   GGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAATCTGATCGCCCAGCTGCCCGGCG 
                 
               
               
                   
               
               
                 
                   AGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAGCCTGGGCCTGACCCCCAACTTCAAGAGC 
                 
               
               
                   
               
               
                 
                   AACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCAAGGACACCTACGACGACGACCTGGACAA 
                 
               
               
                   
               
               
                 
                   CCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTTCTGGCCGCCAAGAACCTGTCCGACGCCA 
                 
               
               
                   
               
               
                 
                   TCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGAGCGCCTCTATGATC 
                 
               
               
                   
               
               
                 
                   AAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGTGCGGCAGCAGCTGCCTGA 
                 
               
               
                   
               
               
                 
                   GAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGCTACGCCGGCTACATTGACGGCGGAGCCA 
                 
               
               
                   
               
               
                 
                   GCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCACCGAGGAACTGCTC 
                 
               
               
                   
               
               
                 
                   GTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAACGGCAGCATCCCCCACCA 
                 
               
               
                   
               
               
                 
                   GATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAGGAAGATTTTTACCCATTCCTGAAGGACA 
                 
               
               
                   
               
               
                 
                   ACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCCCTACTACGTGGGCCCTCTGGCCAGGGGA 
                 
               
               
                   
               
               
                 
                   AACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGGAACTTCGAGGAAGT 
                 
               
               
                   
               
               
                 
                   GGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGGATGACCAACTTCGATAAGAACCTGCCCA 
                 
               
               
                   
               
               
                 
                   ACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTACTTCACCGTGTATAACGAGCTGACCAAA 
                 
               
               
                   
               
               
                 
                   GTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAGAAAAAGGCCATCGT 
                 
               
               
                   
               
               
                 
                   GGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCAGCTGAAAGAGGACTACTTCAAGAAAA 
                 
               
               
                   
               
               
                 
                   TCGAGTGCTTCGACTCCGTGGAAATCTCCGGCGTGGAAGATCGGTTCAACGCCTCCCTGGGCACATAC 
                 
               
               
                   
               
               
                 
                   CACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAACGAGGACATTCTGGA 
                 
               
               
                   
               
               
                 
                   AGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCGAGGAACGGCTGAAAACCTATG 
                 
               
               
                   
               
               
                 
                   CCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCGGCGGAGATACACCGGCTGGGGCAGGCTG 
                 
               
               
                   
               
               
                 
                   AGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTGGATTTCCTGAAGTC 
                 
               
               
                   
               
               
                 
                   CGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACGACAGCCTGACCTTTAAAGAGGACA 
                 
               
               
                   
               
               
                 
                   TCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCACGAGCACATTGCCAATCTGGCCGGCAGC 
                 
               
               
                   
               
               
                 
                   CCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGGTGGACGAGCTCGTGAAAGTGATGGGCCG 
                 
               
               
                   
               
               
                 
                   GCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCCAGAAGGGACAGAAGA 
                 
               
               
                   
               
               
                 
                   ACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAAAGAGCTGuGCAGCCAGATCCTGAAAGAA 
                 
               
               
                   
               
               
                 
                   CACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTGCAGAATGGGCGGGA 
                 
               
               
                   
               
               
                 
                   TATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGACGCCATCGTGCCTC 
                 
               
               
                   
               
               
                 
                   AGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCTGACCAGAAGCGACAAGAACCGGGGCAAG 
                 
               
               
                   
               
               
                 
                   AGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGAAGAACTACTGGCGGCAGCTGCTGAACGC 
                 
               
               
                   
               
               
                 
                   CAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGGCGGCCTGAGCGAACTGG 
                 
               
               
                   
               
               
                 
                   ATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCGGCAGATCACAAAGCACGTGGCACAGATC 
                 
               
               
                   
               
               
                 
                   CTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACAAGCTGATCCGGGAAGTGAAAGTGATCAC 
                 
               
               
                   
               
               
                 
                   CCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGTGCGCGAGATCAACA 
                 
               
               
                   
               
               
                 
                   ACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGTGGGAACCGCCCTGATCAAAAAGTACCCT 
                 
               
               
                   
               
               
                 
                   AAGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGTACGACGTGCGGAAGATGATCGCCAAGAG 
                 
               
               
                   
               
               
                 
                   CGAGCAGGAAATCGGCAAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCATGAACTTTTTCAAGA 
                 
               
               
                   
               
               
                 
                   CCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCCTCTGATCGAGACAAACGGCGAAACCGGG 
                 
               
               
                   
               
               
                 
                   GAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGCGGAAAGTGCTGAGCATGCCCCAAGTGAA 
                 
               
               
                   
               
               
                 
                   TATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAGGAACA 
                 
               
               
                   
               
               
                 
                   GCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCTTCGACAGCCCCACC 
                 
               
               
                   
               
               
                 
                   GTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAACTGAAGAGTGTGAA 
                 
               
               
                   
               
               
                 
                   AGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCATCGACTTTCTGGAAG 
                 
               
               
                   
               
               
                 
                   CCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACTCCCTGTTCGAGCTG 
                 
               
               
                   
               
               
                 
                   GAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAGAAGGGAAACGAACTGGCCCTGCC 
                 
               
               
                   
               
               
                 
                   CTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATA 
                 
               
               
                   
               
               
                 
                   ATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTACCTGGACGAGATCATCGAGCAGATCAGC 
                 
               
               
                   
               
               
                 
                   GAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCCGCCTACAACAAGCA 
                 
               
               
                   
               
               
                 
                   CCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCTGACCAATCTGGGAG 
                 
               
               
                   
               
               
                 
                   CCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCACCAAAGAGGTG 
                 
               
               
                   
               
               
                 
                   CTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGATCGACCTGTCTCAGCT 
                 
               
               
                   
               
               
                 
                   GGGAGGCGAC 
                 
               
               
                   
               
               
                 Amino acid sequence for P300c-dCas9 
               
               
                 (corresponding to SEQ ID NO: 83); lowercase 
               
               
                 underlined = p300c; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 84 
                   
               
               
                 
                   mqifkpeelrqalmptlealyrqdpeslpfrqpvdpqllgipdyfdivkspmdlstikrkldtgqyge 
                 
                   
               
               
                   
               
               
                 
                   pwqyvddiwlmfnnawlynrktsrvykycsklsevfeqeidpvmgslgycogrklefspgtlccyqkq 
                 
               
               
                   
               
               
                 
                   lctiprdatyysygnryhfcekcfneiqgesvslgddpsgpqllinkeqfskrkndlldpelfvecte 
                 
               
               
                   
               
               
                 
                   cgrkmhqicvlhheiiwpagfvcdgcikksartrkenkfsakrlpstrlgtflenrvndflrrqnhpe 
                 
               
               
                   
               
               
                 
                   sqevtvryvhasdktvevkpgmkarfvdsgemaesfpyrtkalfafeeidgvdlcffgmhvqeygsdc 
                 
               
               
                   
               
               
                 
                   pppnqrrvyisyldsvhffrpkclrtavyheiligyleyvkklgyttghiwacppsegddyifhchpp 
                 
               
               
                   
               
               
                 dqkipkpkrlqewykkmldkavserivhdykdifkgatedrltsakelpyfegdfwpnvleesikele 
               
               
                   
               
               
                 
                   qeeeerkreentsnestdvtkgdsknakkknnkktsknkssisrgnkkkpgmpnvsndlsqklvatme 
                 
               
               
                   
               
               
                 
                   khkevffvirliagpaanslppivdpdplipcdlmdgrdafltlardkhlefsslrraqwstmcmlve 
                 
               
               
                   
               
               
                   lhtqsqd gsgasggssat MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGA   
               
               
                   
               
               
                 
                   LLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPI 
                 
               
               
                   
               
               
                 
                   FGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFI 
                 
               
               
                   
               
               
                 
                   QLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKS 
                 
               
               
                   
               
               
                 
                   NFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMI 
                 
               
               
                   
               
               
                 
                   KRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELL 
                 
               
               
                   
               
               
                 
                   VKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG 
                 
               
               
                   
               
               
                 
                   NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTK 
                 
               
               
                   
               
               
                 
                   VKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTY 
                 
               
               
                   
               
               
                 
                   HDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRL 
                 
               
               
                   
               
               
                 
                   SRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGS 
                 
               
               
                   
               
               
                 
                   PAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE 
                 
               
               
                   
               
               
                 
                   HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGK 
                 
               
               
                   
               
               
                 
                   SDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI 
                 
               
               
                   
               
               
                 
                   LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYP 
                 
               
               
                   
               
               
                 
                   KLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETG 
                 
               
               
                   
               
               
                 
                   EIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT 
                 
               
               
                   
               
               
                 
                   VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFEL 
                 
               
               
                   
               
               
                 
                   ENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQIS 
                 
               
               
                   
               
               
                 
                   EFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEV 
                 
               
               
                   
               
               
                 
                   LDATLIHQSITGLYETRIDLSQLGGD 
                 
               
               
                   
               
               
                 DNA sequence for P3000-dCas9 (in backbone pNH 23): pNH32; lowercase 
               
               
                 underlined = p300c; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 85 
                   
               
               
                 
                   atggactacaaagaccatgacggtgattataaagatcatgacatcgattacaaggatgacgatgacaa 
                 
                   
               
               
                   
               
               
                 
                   gcacgttattttcaaaccagaagaactacgacaggcactgatgccaactttggaggcactttaccgtc 
                 
               
               
                   
               
               
                 
                   aggatccagaatcccttccctttcgtcaacctgtggaccatcagcttttaggaatccctgattacttt 
                 
               
               
                   
               
               
                 
                   gatattgtgaagagccccatggatctttctaccattaagaggaagttagacactggacagtatcagga 
                 
               
               
                   
               
               
                 
                   gccctggcagtatgtcgatgatatttggcttatgttcaataatgcctggttatataaccggaaaacat 
                 
               
               
                   
               
               
                 
                   cacgggtatacaaatactgctccaagctctctgaggtctttgaacaagaaattgacccagtgatgcaa 
                 
               
               
                   
               
               
                 
                   agccttggatactgttgtggcagaaagttggagttctctccacagacactgtgttgctacggcaaaca 
                 
               
               
                   
               
               
                 
                   gttgtgcacaatacctcgtgatgccacttattacagttaccagaacaggtatcatttctgtgagaagt 
                 
               
               
                   
               
               
                 
                   gtttcaatgagatccaaggggagagcgtttctttgggggatgacccttcccagcctcaaactacaata 
                 
               
               
                   
               
               
                 
                   aataaagaacaattttccaagagaaaaaatgacacactggatcctgaactgtttgttgaatgtacaga 
                 
               
               
                   
               
               
                 
                   gtgcggaagaaagatgcatcagatctgtgtcattcaccatgagatcatctggcctgctggattcgtct 
                 
               
               
                   
               
               
                 
                   gtgatggctgtttaaagaaaagtgcacgaactaggaaagaaaataagttttctgctaaaaggttgcca 
                 
               
               
                   
               
               
                 
                   tctaccagacttggcacctttctagagaatcgtgtgaatgactttctgaggcgacagaatcaccctga 
                 
               
               
                   
               
               
                 
                   gtcaggagaggtcactgttagagtagttcatgcttctgacaaaaccgtggaagtaaaaccaggcatga 
                 
               
               
                   
               
               
                 
                   aagcaaggtttgtggacagtggagagatggcagaatcctttccataccgaaccaaagccctctttgcc 
                 
               
               
                   
               
               
                 
                   tttgaagaaattgatggtgttgacctgtgcttctttggcatgcatgttcaagagtatggctctgactg 
                 
               
               
                   
               
               
                 
                   ccctccacccaaccagaggagagtatacatatcttacctcgatagtgttcatttcttccgtcctaaat 
                 
               
               
                   
               
               
                 
                   gcttgaggactgcagtctatcatgaaatcctaattggatatttagaatatgtcaagaaattaggttac 
                 
               
               
                   
               
               
                 
                   acaacagggcatatttgggcatgtccaccaagtgagggagatgattatatcttccattgccatcctcc 
                 
               
               
                   
               
               
                 
                   tgaccagaagatacccaagcccaagcgactgcaggaatggtacaaaaaaatgcttgacaaggctgtat 
                 
               
               
                   
               
               
                 
                   cagagcgtattgtccatgactacaaggatatttttaaacaagctactgaagatagattaacaagtgca 
                 
               
               
                   
               
               
                 
                   aaggaattgccttatttcgagggtgatttctggcccaatgttctggaagaaagcattaaggaactgga 
                 
               
               
                   
               
               
                 
                   acaggagaaagaagaaagaaaacgagagaaaaacaccagcaataaaagcacagatgtaaccaaaagag 
                 
               
               
                   
               
               
                 
                   acagcaaaaatgctaaaaagaagaataataagaaaaccagcaaaaataagagcagcctgagtaggggc 
                 
               
               
                   
               
               
                 
                   aacaagaagaaacccgggatgcccaatgtatctaacgacctctcacagaaactatatgccaccatgga 
                 
               
               
                   
               
               
                 
                   gaagcataaagaggtcttctttgtgatccgcctcattgctggccctgctgccaactccctgcctccca 
                 
               
               
                   
               
               
                 
                   ttgttgatcctgatcctctcatcccctgcgatctgatggatggtcgggatgcgtttctcacgctggca 
                 
               
               
                   
               
               
                 
                   agggacaagcacctggagttctcttcactccgaagagcccagtggtccaccatgtgcatgctggtgga 
                 
               
               
                   
               
               
                   gctgcacacgcagagccaggac aaccccaagaagaagaggaaggtgggccgcgga ATGGACAAGAAGT   
               
               
                   
               
               
                 
                   ACTCCATTGGGCTCGCCATCGGCACAAACAGCGTCGGCTGGGCCGTCATTACGGACGAGTACAAGGTG 
                 
               
               
                   
               
               
                 
                   CCGAGCAAAAAATTCAAAGTTCTGGGCAATACCGATCGCCACAGCATAAAGAAGAACCTCATTGGCGC 
                 
               
               
                   
               
               
                 
                   CCTCCTGTTCGACTCCGGGGAAACCGCCGAAGCCACGCGGCTCAAAAGAACAGCACGGCGCAGATATA 
                 
               
               
                   
               
               
                 
                   CCCGCAGAAAGAATCGGATCTGCTACCTGCAGGAGATCTTTAGTAATGAGATGGCTAAGGTGGATGAC 
                 
               
               
                   
               
               
                 
                   TCTTTCTTCCATAGGCTGGAGGAGTCCTTTTTGGTGGAGGAGGATAAAAAGCACGAGCGCCACCCAAT 
                 
               
               
                   
               
               
                 
                   CTTTGGCAATATCGTGGACGAGGTGGCGTACCATGAAAAGTACCCAACCATATATCATCTGAGGAAGA 
                 
               
               
                   
               
               
                 
                   AGCTTGTAGACAGTACTGATAAGGCTGACTTGCGGTTGATCTATCTCGCGCTGGCGCATATGATCAAA 
                 
               
               
                   
               
               
                 
                   TTTCGGGGACACTTCCTCATCGAGGGGGACCTGAACCCAGACAACAGCGATGTCGACAAACTCTTTAT 
                 
               
               
                   
               
               
                 
                   CCAACTGGTTCAGACTTACAATCAGCTTTTCGAAGAGAACCCGATCAACGCATCCGGAGTTGACGCCA 
                 
               
               
                   
               
               
                 
                   AAGCAATCCTGAGCGCTAGGCTGTCCAAATCCCGGCGGCTCGAAAACCTCATCGCACAGCTCCCTGGG 
                 
               
               
                   
               
               
                 GAGAAGAAGAACGGCCTGTTTGGTAATCTTATCGCCCTGTCACTCGGGCTGACCCCCAACTTTAAATC 
               
               
                   
               
               
                 
                   TAACTTCGACCTGGCCGAAGATGCCAAGCTTCAACTGAGCAAAGACACCTACGATGATGATCTCGACA 
                 
               
               
                   
               
               
                 
                   ATCTGCTGGCCCAGATCGGCGACCAGTACGCAGACCTTTTTTTGGCGGCAAAGAACCTGTCAGACGCC 
                 
               
               
                   
               
               
                 
                   ATTCTGCTGAGTGATATTCTGCGAGTGAACACGGAGATCACCAAAGCTCCGCTGAGCGCTAGTATGAT 
                 
               
               
                   
               
               
                 
                   CAAGCGCTATGATGAGCACCACCAAGACTTGACTTTGCTGAAGGCCCTTGTCAGACAGCAACTGCCTG 
                 
               
               
                   
               
               
                 
                   AGAAGTACAAGGAAATTTTCTTCGATCAGTCTAAAAATGGCTACGCCGGATACATTGACGGCGGAGCA 
                 
               
               
                   
               
               
                 
                   AGCCAGGAGGAATTTTACAAATTTATTAAGCCCATCTTGGAAAAAATGGACGGCACCGAGGAGCTGCT 
                 
               
               
                   
               
               
                 
                   GGTAAAGCTTAACAGAGAAGATCTGTTGCGCAAACAGCGCACTTTCGACAATGGAAGCATCCCCCACC 
                 
               
               
                   
               
               
                 
                   AGATTCACCTGGGCGAACTGCACGCTATCCTCAGGCGGCAAGAGGATTTCTACCCCTTTTTGAAAGAT 
                 
               
               
                   
               
               
                 
                   AACAGGGAAAAGATTGAGAAAATCCTCACATTTCGGATACCCTACTATGTAGGCCCCCTCGCCCGGGG 
                 
               
               
                   
               
               
                 
                   AAATTCCAGATTCGCGTGGATGACTCGCAAATCAGAAGAGACCATCACTCCCTGGAACTTCGAGGAAG 
                 
               
               
                   
               
               
                 
                   TCGTGGATAAGGGGGCCTCTGCCCAGTCCTTCATCGAAAGGATGACTAACTTTGATAAAAATCTGCCT 
                 
               
               
                   
               
               
                 
                   AACGAAAAGGTGCTTCCTAAACACTCTCTGCTGTACGAGTACTTCACAGTTTATAACGAGCTCACCAA 
                 
               
               
                   
               
               
                 
                   GGTCAAATACGTCACAGAAGGGATGAGAAAGCGAGCATTCCTGTCTGGAGAGCAGAAGAAAGCTATCG 
                 
               
               
                   
               
               
                 
                   TGGACCTCCTCTTCAAGACGAACCGGAAAGTTACCGTGAAACAGCTCAAAGAAGACTATTTCAAAAAG 
                 
               
               
                   
               
               
                 
                   ATTGAATGTTTCGACTCTGTTGAAATCAGCGGAGTGGAGGATCGCTTCAACGCATCCCTGGGAACGTA 
                 
               
               
                   
               
               
                 
                   TCACGATCTCCTGAAAATCATTAAAGACAAGGACTTCCTGGACAATGAGGAGAACGAGGACATTCTTG 
                 
               
               
                   
               
               
                 
                   AGGACATTGTCCTCACCCTTACGTTGTTTGAAGATAGGGAGATGATTGAAGAACGCTTGAAAACTTAC 
                 
               
               
                   
               
               
                 
                   GCTCATCTCTTCGACGACAAAGTCATGAAACAGCTCAAGAGGCGCCGATATACAGGATGGGGGCGGCT 
                 
               
               
                   
               
               
                 
                   GTCAAGAAAACTGATCAATGGGATCCGAGACAAGCAGAGTGGAAAGACAATCCTGGATTTTCTTAAGT 
                 
               
               
                   
               
               
                 
                   CCGATGGATTTGCCAACCGGAACTTCATGCAGTTGATCCATGATGACTCTCTCACCTTTAAGGAGGAC 
                 
               
               
                   
               
               
                 
                   ATCCAGAAAGCACAAGTTTCTGGCCAGGGGGACAGTCTTCACGAGCACATCGCTAATCTTGCAGGTAG 
                 
               
               
                   
               
               
                 
                   CCCAGCTATCAAAAAGGGAATACTGCAGACCGTTAAGGTCGTGGATGAACTCGTCAAAGTAATGGGAA 
                 
               
               
                   
               
               
                 
                   GGCATAAGCCCGAGAATATCGTTATCGAGATGGCCCGAGAGAACCAAACTACCCAGAAGGGACAGAAG 
                 
               
               
                   
               
               
                 
                   AACAGTAGGGAAAGGATGAAGAGGATTGAAGAGGGTATAATGGAACTGGGGTCCCAAATCCTTAAGGA 
                 
               
               
                   
               
               
                 
                   ACACCCAGTTGAAAACACCCAGCTTCAGAATGAGAAGCTCTACCTGTACTACCTGCAGAACGGCAGGG 
                 
               
               
                   
               
               
                 
                   ACATGTACGTGGATCAGGAACTGGACATCAATCGGCTCTCCGACTACGACGTGGATGCCATCGTGCCC 
                 
               
               
                   
               
               
                 
                   CAGTCTTTTCTCAAAGATGATTCTATTGATAATAAAGTGTTGACAAGATCCGATAAAAATAGAGGGAA 
                 
               
               
                   
               
               
                 
                   GAGTGATAACGTCCCCTCAGAAGAAGTTGTCAAGAAAATGAAAAATTATTGGCGGCAGCTGCTGAACG 
                 
               
               
                   
               
               
                 
                   CCAAACTGATCACACAACGGAAGTTCGATAATCTGACTAAGGCTGAACGAGGTGGCCTGTCTGAGTTG 
                 
               
               
                   
               
               
                 
                   GATAAAGCCGGCTTCATCAAAAGGGAGCTTGTTGAGACACGCCAGATCACCAAGCACGTGGCCCAAAT 
                 
               
               
                   
               
               
                 
                   TCTCGATTCACGCATGAACACCAAGTACGATGAAAATGACAAACTGATTCGAGAGGTGAAAGTTATTA 
                 
               
               
                   
               
               
                 
                   CTCTGAAGTCTAAGCTGGTCTCAGATTTCAGAAAGGACTTTCAGTTTTATAAGGTGAGAGAGATCAAC 
                 
               
               
                   
               
               
                 
                   AATTACCACCATGCGCATGATGCCTACCTGAATGCAGTGGTAGGCACTGCACTTATCAAAAAATATCC 
                 
               
               
                   
               
               
                 
                   CAAGCTTGAATCTGAATTTGTTTACGGAGACTATAAAGTGTACGATGTTAGGAAAATGATCGCAAAGT 
                 
               
               
                   
               
               
                 
                   CTGAGCAGGAAATAGGCAAGGCCACCGCTAAGTACTTCTTTTACAGCAATATTATGAATTTTTTCAAG 
                 
               
               
                   
               
               
                 
                   ACCGAGATTACACTGGCCAATGGAGAGATTCGGAAGCGACCACTTATCGAAACAAACGGAGAAACAGG 
                 
               
               
                   
               
               
                 
                   AGAAATCGTGTGGGACAAGGGTAGGGATTTCGCGACAGTCCGGAAGGTCCTGTCCATGCCGCAGGTGA 
                 
               
               
                   
               
               
                 
                   ACATCGTTAAAAAGACCGAAGTACAGACCGGAGGCTTCTCCAAGGAAAGTATCCTCCCGAAAAGGAAC 
                 
               
               
                   
               
               
                 
                   AGCGACAAGCTGATCGCACGCAAAAAAGATTGGGACCCCAAGAAATACGGCGGATTCGATTCTCCTAC 
                 
               
               
                   
               
               
                 
                   AGTCGCTTACAGTGTACTGGTTGTGGCCAAAGTGGAGAAAGGGAAGTCTAAAAAACTCAAAAGCGTCA 
                 
               
               
                   
               
               
                 
                   AGGAACTGCTGGGCATCACAATCATGGAGCGATCAAGCTTCGAAAAAAACCCCATCGACTTTCTCGAG 
                 
               
               
                   
               
               
                 
                   GCGAAAGGATATAAAGAGGTCAAAAAAGACCTCATCATTAAGCTTCCCAAGTACTCTCTCTTTGAGCT 
                 
               
               
                   
               
               
                 
                   TGAAAACGGCCGGAAACGAATGCTCGCTAGTGCGGGCGAGCTGCAGAAAGGTAACGAGCTGGCACTGC 
                 
               
               
                   
               
               
                 
                   CCTCTAAATACGTTAATTTCTTGTATCTGGCCAGCCACTATGAAAAGCTCAAAGGGTCTCCCGAAGAT 
                 
               
               
                   
               
               
                 
                   AATGAGCAGAAGCAGCTGTTCGTGGAACAACACAAACACTACCTTGATGAGATCATCGAGCAAATAAG 
                 
               
               
                   
               
               
                 
                   CGAATTCTCCAAAAGAGTGATCCTCGCCGACGCTAACCTCGATAAGGTGCTTTCTGCTTACAATAAGC 
                 
               
               
                   
               
               
                 
                   ACAGGGATAAGCCCATCAGGGAGCAGGLAGAAAACATTATCCACTTGTTTACTCTGACCAACTTGGGC 
                 
               
               
                   
               
               
                 
                   GCGCCTGGAGCCTTGAAGTACTTGGACAGCACCATAGACAGAAAGGGGTACACCTCTACAAAGGAGGT 
                 
               
               
                   
               
               
                 
                   CCTGGACGCCACACTGATTCATCAGTCAATTACGGGGCTCTATGAAACAAGAATCGACCTCTCTCAGC 
                 
               
               
                   
               
               
                 
                   TCGGTGGAGACAGCAGGGCTGAC 
                 
               
               
                   
               
               
                 Amino acid sequence for P300c-dCas9 
               
               
                 (corresponding to SEQ ID NO: 85); lowercase 
               
               
                 underlined = p300c; capital underlined = dCas9. 
               
               
                 SEQ ID NO: 86 
                   
               
               
                 dykdhdgdykdhdidykddddkhv ifkpeelrqalmpllealyrqdpeslpfrqpvdpqllqipdyfd   
                   
               
               
                   
               
               
                 
                   ivkspmdlstikrkldtgqyqepwayvddiwlmfnnawlynrktsrvykycsklsevfeqeidpvmgs 
                 
               
               
                   
               
               
                 
                   lgyccgrklefspqtlccygkqlctiprdatyysyqnryhfcekcfneiqgesvslgddpsqpgttin 
                 
               
               
                   
               
               
                 
                   keqfskrkndtldpelfvectecgrkmhqicvlhheiiwpagfvedgclkksartrkenkfsakrlps 
                 
               
               
                   
               
               
                 
                   trigtflenrvndfIrrqnhpesgevtvrvvhasdktvevkpgmkarfvdsgemaesfpyrtkalfaf 
                 
               
               
                   
               
               
                 
                   eeidgvdlcffgmhvqeygsdcpppngrrvyisyldsvhffrpkclrtavyheiligyleyvkklgyt 
                 
               
               
                   
               
               
                 
                   tghiwacppsegddyifhchppdqkipkpkrlqewykkmldkavserivhdykdifkqatedrltsak 
                 
               
               
                   
               
               
                 
                   elpyfegdfwpnvleesikeleqeeeerkreentsnestdvtkgdsknakkknnkktsknksslsrgn 
                 
               
               
                   
               
               
                 
                   kkkpgmpnvsndlsqklyatmekhkevffvirliaqpaanslppivdpdplipcdlmdgrdafltlar 
                 
               
               
                   
               
               
                   dkhlefsslrraqwstmcmlvelhtqsqd npkkkrkvgrg MDKKYSIGLAIGTNSVGWAVITDEYKVP   
               
               
                   
               
               
                 
                   SKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDS 
                 
               
               
                   
               
               
                 
                   FFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF 
                 
               
               
                   
               
               
                 
                   RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGE 
                 
               
               
                   
               
               
                 
                   KKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAI 
                 
               
               
                   
               
               
                 
                   LLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGAS 
                 
               
               
                   
               
               
                 
                   QEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDN 
                 
               
               
                   
               
               
                 
                   REKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPN 
                 
               
               
                   
               
               
                 
                   EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKI 
                 
               
               
                   
               
               
                 
                   ECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYA 
                 
               
               
                   
               
               
                 
                   HLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDI 
                 
               
               
                   
               
               
                 
                   QKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKN 
                 
               
               
                   
               
               
                 
                   SRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQ 
                 
               
               
                   
               
               
                 
                   SFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELD 
                 
               
               
                   
               
               
                 
                   KAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINN 
                 
               
               
                   
               
               
                 
                   YHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT 
                 
               
               
                   
               
               
                 
                   EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNS 
                 
               
               
                   
               
               
                 
                   DKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEA 
                 
               
               
                   
               
               
                 
                   KGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDN 
                 
               
               
                   
               
               
                 
                   EQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA 
                 
               
               
                   
               
               
                 
                   PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSRAD 
                 
               
               
                   
               
               
                 DNA sequence fordCas9-SS18 (in backbone pNI36): 
               
               
                 pNI80; capital underlined = dCas9; 
               
               
                 capital no underline = SS18 
               
               
                 SEQ ID NO: 87 
                   
               
               
                 
                   ATGGACAAGAAGTAGAGCATCGGCCTGGCCATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGA 
                 
                   
               
               
                   
               
               
                 
                   CGAGTACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAAGAAGA 
                 
               
               
                   
               
               
                 
                   ACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCCACCCGGCTGAAGAGAACCGCC 
                 
               
               
                   
               
               
                 
                   AGAAGAAGATACACCAGACGGAAGAACCGGATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGC 
                 
               
               
                   
               
               
                 
                   CAAGGTGGACGACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAAGCACG 
                 
               
               
                   
               
               
                 
                   AGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTAC 
                 
               
               
                   
               
               
                 
                   CACCTGAGAAAGAAACTGGTGGACAGCACCGACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGC 
                 
               
               
                   
               
               
                 
                   CCACATGATCAAGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGG 
                 
               
               
                   
               
               
                 
                   ACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCCCATCAACGCCAGC 
                 
               
               
                   
               
               
                 
                   GGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGCAAGAGCAGACGGCTGGAAAATCTGATCGC 
                 
               
               
                   
               
               
                 
                   CCAGCTGCCCGGCGAGAAGAAGAATGGCCTGTTCGGAAACCTGATTGCCCTGAGCCTGGGCCTGACCC 
                 
               
               
                   
               
               
                 
                   CCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCAAGGACACCTACGAC 
                 
               
               
                   
               
               
                 
                   GACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACGCCGACCTGTTTCTGGCCGCCAAGAA 
                 
               
               
                   
               
               
                 
                   CCTGTCCGACGCCATCCTGCTGAGCGACATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGA 
                 
               
               
                   
               
               
                 
                   GCGCCTCTATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGTGCGG 
                 
               
               
                   
               
               
                 
                   CAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAGAACGGCTACGCCGGCTACAT 
                 
               
               
                   
               
               
                 
                   TGACGGCGGAGCCAGCCAGGAAGAGTTCTACAAGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCA 
                 
               
               
                   
               
               
                 
                   CCGAGGAACTGCTCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAACGGC 
                 
               
               
                   
               
               
                 
                   AGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGGCAGGAAGATTTTTACCC 
                 
               
               
                   
               
               
                 
                   ATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGATCCTGACCTTCCGCATCCCCTACTACGTGGGCC 
                 
               
               
                   
               
               
                 
                   CTCTGGCCAGGGGAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGG 
                 
               
               
                   
               
               
                 
                   AACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGGATGACCAACTTCGA 
                 
               
               
                   
               
               
                 
                   TAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGCCTGCTGTACGAGTACTTCACCGTGTATA 
                 
               
               
                   
               
               
                 
                   ACGAGCTGACCAAAGTGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAG 
                 
               
               
                   
               
               
                 
                   AAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCAGCTGAAAGAGGA 
                 
               
               
                   
               
               
                 
                   CTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCCGGCGTGGAAGATCGGTTCAACGCCT 
                 
               
               
                   
               
               
                 
                   CCCTGGGCACATACCACGATCTGCTGAAAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAAC 
                 
               
               
                   
               
               
                 
                   GAGGACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCGAGGAACG 
                 
               
               
                   
               
               
                 
                   GCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCGGCGGAGATACACCG 
                 
               
               
                   
               
               
                 
                   GCTGGGGCAGGCTGAGCCGGAAGCTGATCAACGGCATCCGGGACAAGCAGTCCGGLAAGACAATCCTG 
                 
               
               
                   
               
               
                 
                   GATTTCCTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACGACAGCCTGAC 
                 
               
               
                   
               
               
                 
                   CTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCCTGCACGAGCACATTGCCA 
                 
               
               
                   
               
               
                 
                   ATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCATCCTGCAGACAGTGAAGGTGGTGGACGAGCTCGTG 
                 
               
               
                   
               
               
                 
                   AAAGTGATGGGCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCCA 
                 
               
               
                   
               
               
                 
                   GAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGCATCAAAGAGCTGGGCAGCC 
                 
               
               
                   
               
               
                 
                   AGATCCTGAAAGAACACCCCGTGGAAAACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTG 
                 
               
               
                   
               
               
                 
                   CAGAATGGGCGGGATATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGA 
                 
               
               
                   
               
               
                 
                   CGCCATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCTGACCAGAAGCGACA 
                 
               
               
                   
               
               
                 
                   AGAACCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAGGTCGTGAAGAAGATGAAGAACTACTGGCGG 
                 
               
               
                   
               
               
                 
                   CAGCTGCTGAACGCCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGGCGG 
                 
               
               
                   
               
               
                 
                   CCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCGGCAGATCACAAAGC 
                 
               
               
                   
               
               
                 
                   ACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACGACGAGAATGACAAGCTGATCCGGGAA 
                 
               
               
                   
               
               
                 
                   GTGAAAGTGATCACCCTGAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGT 
                 
               
               
                   
               
               
                 
                   GCGCGAGATCAACAACTAGCACCACGCCCACGACGCCTAGCTGAACGCCGTCGTGGGAACCGCCCTGA 
                 
               
               
                   
               
               
                 TCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTACAAGGTGTACGACGTGCGGAAG 
               
               
                   
               
               
                 
                   ATGATCGCCAAGAGCGAGCAGGAAATCGGCAAGuCTACCGCCAAGTACTTCTTCTACAGCAACATCAT 
                 
               
               
                   
               
               
                 
                   GAACTTTTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCCTCTGATCGAGACAA 
                 
               
               
                   
               
               
                 
                   ACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCCACCGTGCGGAAAGTGCTGAGC 
                 
               
               
                   
               
               
                 
                   ATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCT 
                 
               
               
                   
               
               
                 
                   GCCCAAGAGGAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCT 
                 
               
               
                   
               
               
                 
                   TCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAAAGGGCAAGTCCAAGAAA 
                 
               
               
                   
               
               
                 
                   CTGAAGAGTGTGAAAGAGCTGCTGGGGATCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCAT 
                 
               
               
                   
               
               
                 
                   CGACTTTCTGGAAGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACT 
                 
               
               
                   
               
               
                 
                   CCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAGAAGGGAAAC 
                 
               
               
                   
               
               
                 
                   GAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGGCCAGCCACTATGAGAAGCTGAAGGG 
                 
               
               
                   
               
               
                 
                   CTCCCCCGAGGATAATGAGCAGAAACAGCTGTTTGTGGAACAGCACAAGCACTACCTGGACGAGATCA 
                 
               
               
                   
               
               
                 
                   TCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGCTGTCC 
                 
               
               
                   
               
               
                 
                   GCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGAATATCATCCACCTGTTTACCCT 
                 
               
               
                   
               
               
                 
                   GACCAATCTGGGAGCCCCTGCCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCA 
                 
               
               
                   
               
               
                 
                   GCACCAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGATC 
                 
               
               
                   
               
               
                   GACCTGTCTCAGCTGGGAGGCGAC aagcgacctgccgccacaaagaaggctgqacaggctaagaagaa 
               
               
                   
               
               
                 gaaactggactctggaggatccgactacaaagaccatgacggtgattataaagatcatgacatcgatt 
               
               
                   
               
               
                 acaaggatgacgatgacaagggaggatccaaggagaagagtgcttgtcctaaagatccagccaaacct 
               
               
                   
               
               
                 ccggccaaggcacaagttgtgggatggccaccggtgagatcataccggaagaacgtgatggtttcctg 
               
               
                   
               
               
                 ccaaaaatcaagcggtggcccggaggcggcggcgttcgtgaaggtatcaatggacggagcaccgtact 
               
               
                   
               
               
                 tgaggaaaatcgatttgaggatgtataaaggcggatctggcggctctggaggatccagcATGTCTGTG 
               
               
                   
               
               
                 GCTTTCGCGGCCCCGAGGCAGCGAGGCAAGGGGGAGATCACTCCCGCTGCGATTCAGAAGATGTTGGA 
               
               
                   
               
               
                 TGACAATAACCATCTTATTCAGTGTATAATGGACTCTCAGAATAAAGGAAAGACCTCAGAGTGTTCTC 
               
               
                   
               
               
                 AGTATCAGCAGATGTTGCACACAAACTTGGTATACCTTGCTACAATAGCAGATTCTAATCAAAATATG 
               
               
                   
               
               
                 CAGTCTCTTTTACCAGCACCACCCACACAGAATATGCCTATGGGTCCTGGAGGGATGAATCAGAGCGG 
               
               
                   
               
               
                 CCCTCCCCCACCTCCACGCTCTCACAACATGCCTTCAGATGGAATGGTAGGTGGGGGTCCTCCTGCAC 
               
               
                   
               
               
                 CGCACATGCAGAACCAGATGAACGGCCAGATGCCTGGGCCTAACCATATGCCTATGCAGGGACCTGGA 
               
               
                   
               
               
                 CCCAATCAACTCAATATGACAAACAGTTCCATGAATATGCCTTCAAGTAGCCATGGATCCATGGGAGG 
               
               
                   
               
               
                 TTACAACCATTCTGTGCCATCATCACAGAGCATGCCAGTACAGAATCAGATGACAATGAGTCAGGGAC 
               
               
                   
               
               
                 AACCAATGGGAAACTATGGTCCCAGACCAAATATGAGTATGCAGCCAAACCAAGGTCCAATGATGCAT 
               
               
                   
               
               
                 CAGCAGCCTCCTTCTCAGCAATACAATATGCCACAGGGAGGCGGACAGCATTACCAAGGACAGCAGCC 
               
               
                   
               
               
                 ACCTATGGGAATGATGGGTCAAGTTAACCAAGGCAATCATATGATGGGTCAGAGACAGATTCCTCCCT 
               
               
                   
               
               
                 ATAGACCTCCTCAACAGGGCCCACCACAGCAGTACTCAGGCCAGGAAGACTATTACGGGGACCAATAC 
               
               
                   
               
               
                 AGTCATGGTGGACAAGGTCCTCCAGAAGGCATGAACCAGCAATATTACCCTGATGGAAATTCACAGTA 
               
               
                   
               
               
                 TGGCCAACAGCAAGATGCATACCAGGGACCACCTCCACAACAGGGATATCCACCCCAGCAGCAGCAGT 
               
               
                   
               
               
                 ACCCAGGGCAGCAAGGTTACCCAGGACAGCAGCAGGGCTACGGTCCTTCACAGGGTGGTCCAGGTCCT 
               
               
                   
               
               
                 CAGTATCCTAACTACCCACAGGGACAAGGTCAGCAGTATGGAGGATATAGACCAACACAGCCTGGACC 
               
               
                   
               
               
                 ACCACAGCCACCCCAGCAGAGGCCTTATGGATATGACCAGGGACAGTATGGAAATTACCAGCAGTGA 
               
               
                   
               
               
                 Amino acid sequence fordCas9-SS18 
               
               
                 (corresponding to SEQ ID NO: 87); capital 
               
               
                 underlined = dCas9; capital no underline = SS18. 
               
               
                 SEQ ID NO: 88 
                   
               
               
                 
                   MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                 
                   
               
               
                   
               
               
                 
                   RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIY 
                 
               
               
                   
               
               
                 
                   HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
                 
               
               
                   
               
               
                 
                   GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
                 
               
               
                   
               
               
                 
                   DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
                 
               
               
                   
               
               
                 
                   QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
                 
               
               
                   
               
               
                 
                   SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
                 
               
               
                   
               
               
                 
                   NFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQ 
                 
               
               
                   
               
               
                 
                   KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
                 
               
               
                   
               
               
                 
                   EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
                 
               
               
                   
               
               
                 
                   DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
                 
               
               
                   
               
               
                 
                   KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
                 
               
               
                   
               
               
                 
                   QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
                 
               
               
                   
               
               
                 
                   QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
                 
               
               
                   
               
               
                 
                   VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK 
                 
               
               
                   
               
               
                 
                   MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
                 
               
               
                   
               
               
                 
                   MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLvvAKVEKGKSKK 
                 
               
               
                   
               
               
                 
                   LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
                 
               
               
                   
               
               
                 
                   ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLS 
                 
               
               
                   
               
               
                 
                   AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
                 
               
               
                   
               
               
                   DLSQLGGD krpaatkkagqakkkkidsggsdykdhdgdykdhdidykddddkggskeksacpkdpakp 
               
               
                   
               
               
                 pakaqvvgwppvrsyrknvmvscqkssggpeaaafvkvsmdgapylrkidlrmykggsggsggssMSV 
               
               
                   
               
               
                 AFAAPRQRGKGEITPAAIQKMLDDNNHLIQCIMDSQNKGKTSECSQYQQMLHTNLVYLATIADSNQNM 
               
               
                   
               
               
                 QSLLPAPPTQNMPMGPGGMNQSGPPPPPRSHNMPSDGMVGGGPPAPHMQNQMNGQMPGPNEMPMQGPG 
               
               
                   
               
               
                 PNQLNMTNSSMNMPSSSHGSMGGYNHSVPSSQSMPVQNQMTMSQGQPMGNYGPRPNMSMQPNQGPMMH 
               
               
                   
               
               
                 QQPPSQQYNMPQGGGQHYQGQQPPMGMMGQVNQGNHMMGQRQIPPYRPPQQGPPQQYSGQEDYYGDQY 
               
               
                   
               
               
                 SHGGQGPPEGMNQQYYPDGNSQYGQQQDAYQGPPPQQGYPPQQQQYPGQQGYPGQQQGYGPSQGGPGP 
               
               
                   
               
               
                 QYPNYPQGQGQQYGGYRPTQPGPPQPPQQRPYGYDQGQYGNYQQ* 
               
               
                   
               
               
                 DNA sequence fordCas9-SS18 (in backbone pNH 44): pNi228; 
               
               
                 capital underlined = dCas9; 
               
               
                 capital no underline = SS18. 
               
               
                 SEQ ID NO: 89 
                   
               
               
                 
                   ATGGACAAGAAGTACTCCATTGGGCTCGCCATCGGCACAAACAGCGTCGGCTGGGCCGTCATTACGGA 
                 
                   
               
               
                   
               
               
                 
                   CGAGTACAAGGTGCCGAGCAAAAAATTCAAAGTTCTGGGCAATACCGATCGCCACAGCATAAAGAAGA 
                 
               
               
                   
               
               
                 
                   ACCTCATTGGCGCCCTCCTGTTCGACTCCGGGGAAACCGCCGAAGCCACGCGGCTCAAAAGAACAGCA 
                 
               
               
                   
               
               
                 
                   CGGCGCAGATATACCCGCAGAAAGAATCGGATCTGCTACCTGCAGGAGATCTTTAGTAATGAGATGGC 
                 
               
               
                   
               
               
                 
                   TAAGGTGGATGACTCTTTCTTCCATAGGCTGGAGGAGTCCTTTTTGGTGGAGGAGGATAAAAAGCACG 
                 
               
               
                   
               
               
                 
                   AGCGCCACCCAATCTTTGGCAATATCGTGGACGAGGTGGCGTACCATGAAAAGTACCCAACCATATAT 
                 
               
               
                   
               
               
                 
                   CATCTGAGGAAGAAGCTTGTAGACAGTACTGATAAGGCTGACTTGCGGTTGATCTATCTCGCGCTGGC 
                 
               
               
                   
               
               
                 
                   GCATATGATCAAATTTCGGGGACACTTCCTCATCGAGGGGGACCTGAACCCAGACAACAGCGATGTCG 
                 
               
               
                   
               
               
                 
                   ACAAACTCTTTATCCAACTGGTTCAGACTTACAATCAGCTTTTCGAAGAGAACCCGATCAACGCATCC 
                 
               
               
                   
               
               
                 
                   GGAGTTGACGCCAAAGCAATCCTGAGCGCTAGGCTGTCCAAATCCCGGCGGCTCGAAAACCTCATCGC 
                 
               
               
                   
               
               
                 
                   ACAGCTCCCTGGGGAGAAGAAGAACGGCCTGTTTGGTAATCTTATCGCCCTGTCACTCGGGCTGACCC 
                 
               
               
                   
               
               
                 
                   CCAACTTTAAATCTAACTTCGACCTGGCCGAAGATGCCAAGCTTCAACTGAGCAAAGACACCTACGAT 
                 
               
               
                   
               
               
                 
                   GATGATCTCGACAATCTGCTGGCCCAGATCGGCGACCAGTACGCAGACCTTTTTTTGGCGGCAAAGAA 
                 
               
               
                   
               
               
                 
                   CCTGTCAGACGCCATTCTGCTGAGTGATATTCTGCGAGTGAACACGGAGATCACCAAAGCTCCGCTGA 
                 
               
               
                   
               
               
                 
                   GCGCTAGTATGATCAAGCGCTATGATGAGCACCACCAAGACTTGACTTTGCTGAAGGCCCTTGTCAGA 
                 
               
               
                   
               
               
                 
                   CAGCAACTGCCTGAGAAGTACAAGGAAATTTTCTTCGATCAGTCTAAAAATGGCTACGCCGGATACAT 
                 
               
               
                   
               
               
                 
                   TGACGGCGGAGCAAGCCAGGAGGAATTTTACAAATTTATTAAGCCCATCTTGGAAAAAATGGACGGCA 
                 
               
               
                   
               
               
                 
                   CCGAGGAGCTGCTGGTAAAGCTTAACAGAGAAGATCTGTTGCGCAAACAGCGCACTTTCGACAATGGA 
                 
               
               
                   
               
               
                 
                   AGCATCCCCCACCAGATTCACCTGGGCGAACTGCACGCTATCCTCAGGCGGCAAGAGGATTTCTACCC 
                 
               
               
                   
               
               
                 
                   CTTTTTGAAAGATAACAGGGAAAAGATTGAGAAAATCCTCACATTTCGGATACCCTACTATGTAGGCC 
                 
               
               
                   
               
               
                 
                   CCCTCGCCCGGGGAAATTCCAGATTCGCGTGGATGACTCGCAAATCAGAAGAGACCATCACTCCCTGG 
                 
               
               
                   
               
               
                 
                   AACTTCGAGGAAGTCGTGGATAAGGGGGCCTCTGCCCAGTCCTTCATCGAAAGGATGACTAACTTTGA 
                 
               
               
                   
               
               
                 
                   TAAAAATCTGCCTAACGAAAAGGTGCTTCCTAAACACTCTCTGCTGTACGAGTACTTCACAGTTTATA 
                 
               
               
                   
               
               
                 
                   ACGAGCTCACCAAGGTCAAATACGTCACAGAAGGGATGAGAAAGCCAGCATTCCTGTCTGGAGAGCAG 
                 
               
               
                   
               
               
                 
                   AAGAAAGCTATCGTGGACCTCCTCTTCAAGACGAACCGGAAAGTTACCGTGAAACAGCTCAAAGAAGA 
                 
               
               
                   
               
               
                 
                   CTATTTCAAAAAGATTGAATGTTTCGACTCTGTTGAAATCAGCGGAGTGGAGGATCGCTTCAACGCAT 
                 
               
               
                   
               
               
                 
                   CCCTGGGAACGTATCACGATCTCCTGAAAATCATTAAAGACAAGGACTTCCTGGACAATGAGGAGAAC 
                 
               
               
                   
               
               
                 
                   GAGGACATTCTTGAGGACATTGTCCTCACCCTTACGTTGTTTGAAGATAGGGAGATGATTGAAGAACG 
                 
               
               
                   
               
               
                 
                   CTTGAAAACTTACGCTCATCTCTTCGACGACAAAGTCATGAAACAGCTCAAGAGGCGCCGATATACAG 
                 
               
               
                   
               
               
                 
                   GATGGGGGCGGCTGTCAAGAAAACTGATCAATGGGATCCGAGACAAGCAGAGTGGAAAGACAATCCTG 
                 
               
               
                   
               
               
                 
                   GATTTTCTTAAGTCCGATGGATTTGCCAACCGGAACTTCATGCAGTTGATCCATGATGACTCTCTCAC 
                 
               
               
                   
               
               
                 
                   CTTTAAGGAGGACATCCAGAAAGCACAAGTTTCTGGCCAGGGGGACAGTCTTCACGAGCACATCGCTA 
                 
               
               
                   
               
               
                 
                   ATCTTGCAGGTAGCCCAGCTATCAAAAAGGGAATACTGCAGACCGTTAAGGTCGTGGATGAACTCGTC 
                 
               
               
                   
               
               
                 
                   AAAGTAATGGGAAGGCATAAGCCCGAGAATATCGTTATCGAGATGGCCCGAGAGAACCAAACTACCCA 
                 
               
               
                   
               
               
                 
                   GAAGGGACAGAAGAACAGTAGGGAAAGGATGAAGAGGATTGAAGAGGGTATAAAAGAACTGGGGTCCC 
                 
               
               
                   
               
               
                 
                   AAATCCTTAAGGAACACCCAGTTGAAAACACCCAGCTTCAGAATGAGAAGCTCTACCTGTACTACCTG 
                 
               
               
                   
               
               
                 
                   CAGAACGGCAGGGACATGTACGTGGATCAGGAACTGGACATCAATCGuCTCTCCGACTACGACGTGGA 
                 
               
               
                   
               
               
                 
                   TGCCATCGTGCCCCAGTCTTTTCTCAAAGATGATTCTATTGATAATAAAGTGTTGACAAGATCCGATA 
                 
               
               
                   
               
               
                 
                   AAAATAGAGGGAAGAGTGATAACGTCCCCTCAGAAGAAGTTGTCAAGAAAATGAAAAATTATTGGCGG 
                 
               
               
                   
               
               
                 
                   CAGCTGCTGAACGCCAAACTGATCACACAACGGAAGTTCGATAATCTGACTAAGGCTGAACGAGGTGG 
                 
               
               
                   
               
               
                 
                   CCTGTCTGAGTTGGATAAAGCCGGCTTCATCAAAAGGCAGCTTGTTGAGACACGCCAGATCACCAAGC 
                 
               
               
                   
               
               
                 
                   ACGTGGCCCAAATTCTCGATTCACGCATGAACACCAAGTACGATGAAAATGACAAACTGATTCGAGAG 
                 
               
               
                   
               
               
                 
                   GTGAAAGTTATTACTCTGAAGTCTAAGCTGGTCTCAGATTTGAGAAAGGACTTTGAGTTTTATAAGGT 
                 
               
               
                   
               
               
                 
                   GAGAGAGATCAACAATTACCACCATGCGCATGATGCCTACCTGAATGCAGTGGTAGGCACTGCACTTA 
                 
               
               
                   
               
               
                 
                   TCAAAAAATATCCCAAGCTTGAATCTGAATTTGTTTACGGAGACTATAAAGTGTACGATGTTAGGAAA 
                 
               
               
                   
               
               
                 
                   ATGATCGCAAAGTCTGAGCAGGAAATAGGCAAGGCCACCGCTAAGTACTTCTTTTACAGCAATATTAT 
                 
               
               
                   
               
               
                 
                   GAATTTTTTCAAGACCGAGATTACACTGGCCAATGGAGAGATTCGGAAGCGACCACTTATCGAAACAA 
                 
               
               
                   
               
               
                 
                   ACGGAGAAACAGGAGAAATCGTGTGGGACAAGGGTAGGGATTTCGCGACAGTCCGGAAGGTCCTGTCC 
                 
               
               
                   
               
               
                 
                   ATGCCGCAGGTGAACATCGTTAAAAAGACCGAAGTACAGACCGGAGGCTTCTCCAAGGAAAGTATCCT 
                 
               
               
                   
               
               
                 
                   CCCGAAAAGGAACAGCGACAAGCTGATCGCACGCAAAAAAGATTGGGACCCCAAGAAATACGGCGGAT 
                 
               
               
                   
               
               
                 
                   TCGATTCTCCTACAGTCGCTTACAGTGTACTGGTTGTGGCCAAAGTGGAGAAAGGGAAGTCTAAAAAA 
                 
               
               
                   
               
               
                 
                   CTCAAAAGCGTCAAGGAACTGCTGGGCATCACAATCATGGAGCGATCAAGCTTCGAAAAAAACCCCAT 
                 
               
               
                   
               
               
                 
                   CGACTTTCTCGAGGCGAAAGGATATAAAGAGGTCAAAAAAGACCTCATCATTAAGCTTCCCAAGTACT 
                 
               
               
                   
               
               
                 
                   CTCTCTTTGAGCTTGAAAACGGCCGGAAACGAATGCTCGCTAGTGCGGGCGAGCTGCAGAAAGGTAAC 
                 
               
               
                   
               
               
                 
                   GAGCTGGCACTGCCCTCTAAATACGTTAATTTCTTGTATCTGGCCAGCCACTATGAAAAGCTCAAAGG 
                 
               
               
                   
               
               
                 
                   GTCTCCCGAAGATAATGAGCAGAAGCAGCTGTTCGTGGAACAACACAAACACTACCTTGATGAGATCA 
                 
               
               
                   
               
               
                 
                   TCGAGCAAATAAGCGAATTCTCCAAAAGAGTGATCCTCGCCGACGCTAACCTCGATAAGGTGCTTTCT 
                 
               
               
                   
               
               
                 
                   GCTTACAATAAGCACAGGGATAAGCCCATCAGGGAGCAGGCAGAAAACATTATCCACTTGTTTACTCT 
                 
               
               
                   
               
               
                 GACCAACTTGGGCGCGCCTGCAGCCTTCAAGTACTTCGACACCACCATAGACAGAAAGCGGTACACCT 
               
               
                   
               
               
                 
                   CTACAAAGGAGGTCCTGGACGCCACACTGATTCATCAGTCAATTACGGGGCTCTATGAAACAAGAATC 
                 
               
               
                   
               
               
                   GACCTCTCTCAGCTCGGTGGAGACAGCAGGGCTGAC cccaagaagaagaggaaggtggctagCATGTC 
               
               
                   
               
               
                 TGTGGCTTTCGCGGCCCCGAGGCAGCGAGGCAAGGGGGAGATCACTCCCGCTGCGATTCAGAAGATGT 
               
               
                   
               
               
                 TGGATGACAATAACCATCTTATTCAGTGTATAATGGACTCTCAGAATAAAGGAAAGACCTCAGAGTGT 
               
               
                   
               
               
                 TCTCAGTATCAGCAGATGTTGCACACAAACTTGGTATACCTTGCTACAATAGCAGATTCTAATCAAAA 
               
               
                   
               
               
                 TATGCAGTCTCTTTTACCAGCACCACCCACACAGAATATGCCTATGGGTCCTGGAGGGATGAATCAGA 
               
               
                   
               
               
                 GCGGCCCTCCCCCACCTCCACGCTCTCACAACATGCCTTCAGATGGAATGGTAGGTGGGGGTCCTCCT 
               
               
                   
               
               
                 GCACCGCACATGCAGAACCAGATGAACGGCCAGATGCCTGGGCCTAACCATATGCCTATGCAGGGACC 
               
               
                   
               
               
                 TGGACCCAATCAACTCAATATGACAAACAGTTCCATGAATATGCCTTCAAGTAGCCATGGATCCATGG 
               
               
                   
               
               
                 GAGGTTAGAACCATTCTGTGCCATCATCACAGAGCATGCCAGTACAGAATCAGATGACAATGAGTCAG 
               
               
                   
               
               
                 GGACAACCAATGGGAAACTATGGTCCCAGACCAAATATGAGTATGCAGCCAAACCAAGGTCCAATGAT 
               
               
                   
               
               
                 GCATCAGCAGCCTCCTTCTCAGCAATACAATATGCCACAGGGAGGCGGACAGCATTACCAAGGACAGC 
               
               
                   
               
               
                 AGCCACCTATGGGAATGATGGGTCAAGTTAACCAAGGCAATCATATGATGGGTCAGAGACAGATTCCT 
               
               
                   
               
               
                 CCCTATAGACCTCCTCAACAGGGCCCACCACAGCAGTACTCAGGCCAGGAAGACTATTACGGGGACCA 
               
               
                   
               
               
                 ATACAGTCATGGTGGACAAGGTCCTCCAGAAGGCATGAACCAGCAATATTACCCTGATGGAAATTCAC 
               
               
                   
               
               
                 AGTATGGCCAACAGCAAGATGCATACCAGGGACCACCTCCACAACAGGGATATCCACCCCAGCAGCAG 
               
               
                   
               
               
                 CAGTACCCAGGGCAGCAAGGTTACCCAGGACAGCAGCAGGGCTACGGTCCTTCACAGGGTGGTCCAGG 
               
               
                   
               
               
                 TCCTCAGTATCCTAACTACCCACAGGGACAAGGTCAGCAGTATGGAGGATATAGACCAACACAGCCTG 
               
               
                   
               
               
                 GACCACCACAGCCACCCCAGCAGAGGCCTTATGGATATGACCAGGGACAGTATGGAAATTACCAGCAG 
               
               
                   
               
               
                 Amino acid sequence fordCas9-SS18 
               
               
                 (corresponding to SEQ ID NO: 89); capital 
               
               
                 underllned = dCas9; capital no underline = SS18. 
               
               
                 SEQ ID NO: 90 
                   
               
               
                 
                   MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTA 
                 
                   
               
               
                   
               
               
                 
                   RRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIY 
                 
               
               
                   
               
               
                 
                   HLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS 
                 
               
               
                   
               
               
                 
                   GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYD 
                 
               
               
                   
               
               
                 
                   DDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR 
                 
               
               
                   
               
               
                 
                   QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG 
                 
               
               
                   
               
               
                 
                   SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPW 
                 
               
               
                   
               
               
                 
                   NFEEVVDKGASAQSFTERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQ 
                 
               
               
                   
               
               
                 
                   KKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEEN 
                 
               
               
                   
               
               
                 
                   EDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTIL 
                 
               
               
                   
               
               
                 
                   DFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELV 
                 
               
               
                   
               
               
                 
                   KVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL 
                 
               
               
                   
               
               
                 
                   QNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWR 
                 
               
               
                   
               
               
                 
                   QLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIRE 
                 
               
               
                   
               
               
                 
                   VKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAvvGTALIKKYPKLESEFVYGDYKVYDVRK 
                 
               
               
                   
               
               
                 
                   MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS 
                 
               
               
                   
               
               
                 
                   MPQVNTVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVARVEKGKSKK 
                 
               
               
                   
               
               
                 
                   LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN 
                 
               
               
                   
               
               
                 
                   ELALPSKYVNFLYLASHYEKTKGSPEDNFOKOLFVFOHKHYLDETTEQJSEFSKRVILADANLDKVLS 
                 
               
               
                   
               
               
                 
                   AYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRI 
                 
               
               
                   
               
               
                   DLSQLGGDSRAD pkkkrkvasMSVAFAAPRQRGKGEITPAAIQKMLDDNNHLIQCIMDSQNKGKTSEC 
               
               
                   
               
               
                 SQYQQMLHTNLVYLATIADSNQNMQSLLPAPPTQNMPMGPGGMNQSGPPPPPRSHNMPSDGMVGGGPP 
               
               
                   
               
               
                 APHMQNQMNGQMPGPNHMPMQGPGPNQLNMTNSSMNMPSSSHGSMGGYNHSVPSSQSMPVQNQMTMSQ 
               
               
                   
               
               
                 GOPMGNYGPRPNMSMOPNQGPMMHQQPPSOQYNMPQGGGQHYQGOQPPMGMMGQVNQGNHMMGQROIP 
               
               
                   
               
               
                 PYRPPQQGPPQQYSGQEDYYGDQYSHGGQGPPEGMNQQYYPDGNSQYGQQQDAYQGPPPQQGYPPQQQ 
               
               
                   
               
               
                 QYPGQQGYPGQQQGYGPSQGGPGPQYPNYPQGQGQQYGGYRPTQPGPPQPPQQRPYGYDQGQYGNYQQ 
               
               
                   
               
               
                 Amino acid sequence of VP64 
               
               
                 SEQ ID NO: 91 
                   
               
               
                 RADALDDFDLDMLGSDALDDFDLDHLGSDALDDFDLDMLGSDALDDFDLDM 
                   
               
               
                   
               
               
                 DNA sequence of VP64 
               
               
                 SEQ ID NO: 92 
                   
               
               
                 cgggctgacgcattggacgattttgatctggatatgctgggaagtgacgccctcgatgattttgacct 
                   
               
               
                   
               
               
                 tgacatgcttggttcggatgcccttgatgactttgacctcgacatgctcggcagtgacgcccttgatg 
               
               
                   
               
               
                 atttcgacctggacatg 
               
               
                   
               
               
                 Polypeptide sequence of Tet1CD 
               
               
                 SEQ ID NO: 93 
                   
               
               
                 LPTCSCLDRVIQKDKGPYYTHLGAGPSVAAVREIMENRYGQKGNAIRIEIVVYTGKEGKSSHGCPIAK 
                   
               
               
                   
               
               
                 WVLRRSSDEEKVLCLVRQRTGHHCPTAVMVVLIMVWDGIPLPMADRLYTELTENLKSYNGHPTDRRCT 
               
               
                   
               
               
                 LNENRTCTCQGIDPETCGASFSFGCSWSMYFNGCKFGRSPSPRRFRIDPSSPLHEKNLEDNLQSLATR 
               
               
                   
               
               
                 LAPIYKQYAPVAYQNQVEYENVARECRLGSKEGRPFSGVTACLDFCAHPHRDIHNMNNGSTVVCTLTR 
               
               
                   
               
               
                 EDNRSLGVIPODEQLHVLPLYKLSDTDEFGSKEGMEAKIKSGAIEVLAPRRKKRTCFTQPVPRSGKKR 
               
               
                   
               
               
                 AAMMTEVLAHKIRAVEKKPIPRIKRKNNSTTTNNSKPSSLPTLGSNTETVQPEVKSETEPHFILKSSD 
               
               
                   
               
               
                 NTKTYSLMPSAPHPVKEASPGFSWSPKTASATPAPLKNDATASCGFSERSSTPHCTMPSGRLSGANAA 
               
               
                   
               
               
                 AADGPGISQLGEVAPLPTLSAPVMEPLINSEPSTGVTEPLTPHQPNHQPSFLTSPQDLASSPMEEDEQ 
               
               
                   
               
               
                 HSEADEPPSDEPLSDDPLSPAEEKLPHIDEYWSDSEHIFLDANIGGVAIAPAHGSVLIECARRELHAT 
               
               
                   
               
               
                 TPVEHPNRNHPTRLSLVFYQHKNLNKPQHGFELNKIKFEAKEAKNKKMKASEQKDQAANEGPEQSSEV 
               
               
                   
               
               
                 NELNQIPSHKALTLTHDNVVTVSPYALTHVAGPYNHWV 
               
               
                   
               
               
                 Polynucleotide sequence of Tet1CD 
               
               
                 SEQ ID NO: 94 
                   
               
               
                 CTGCCCACCTGCAGCTGTCTTGATCGAGTTATACAAAAAGACAAAGGCCCATATTATACACACCTTGG 
                   
               
               
                   
               
               
                 GGCAGGACCAAGTGTTGCTGCTGTCAGGGAAATCATGGAGAATAGGTATGGTCAAAAAGGAAACGCAA 
               
               
                   
               
               
                 TAAGGATAGAAATAGTAGTGTACACCGGTAAAGAAGGGAAAAGCTCTCATGGGTGTCCAATTGCTAAG 
               
               
                   
               
               
                 TGGGTTTTAAGAAGAAGCAGTGATGAAGAAAAAGTTCTTTGTTTGGTCCGGCAGCGTACAGGCCACCA 
               
               
                   
               
               
                 CTGTCCAACTGCTGTGATGGTGGTGCTCATCATGGTGTGGGATGGCATCCCTCTTCCAATGGCCGACC 
               
               
                   
               
               
                 GGCTATACACAGAGCTCACAGAGAATCTAAAGTCATACAATGGGCACCCTACCGACAGAAGATGCACC 
               
               
                   
               
               
                 CTCAATGAAAATCGTACCTGTACATGTCAAGGAATTGATCCAGAGACTTGTGGAGCTTCATTCTCTTT 
               
               
                   
               
               
                 TGGCTGTTCATGGAGTATGTACTTTAATGGCTGTAAGTTTGGTAGAAGCCCAAGCCCCAGAAGATTTA 
               
               
                   
               
               
                 GAATTGATCCAAGCTCTCCCTTAGATGAAAAAAACCTTGAAGATAACTTACAGAGTTTGGCTAGACGA 
               
               
                   
               
               
                 TTAGCTCCAATTTATAAGCAGTATGCTCCAGTAGCTTACCAAAATCAGGTGGAATATGAAAATGTTGC 
               
               
                   
               
               
                 CCGAGAATGTCGGCTTGGCAGCAAGGAAGGTCGACCCTTCTCTGGGGTCACTGCTTGCCTGGACTTCT 
               
               
                   
               
               
                 GTGCTCATCCCCACAGGGACATTCACAACATGAATAATGGAAGCACTGTGGTTTGTACCTTAACTCGA 
               
               
                   
               
               
                 GAAGATAACCGCTCTTTGGGTGTTATTCCTCAAGATGAGCAGCTCCATGTGCTACCTCTTTATAAGCT 
               
               
                   
               
               
                 TTCAGACACAGATGAGTTTGGCTCCAAGGAAGGAATGGAAGCCAAGATCAAATCTGGGGCCATCGAGG 
               
               
                   
               
               
                 TCCTGGCACCCCGCCGCAAAAAAAGAACGTGTTTCACTCAGCCTGTTCCCCGTTCTGGAAAGAAGAGG 
               
               
                   
               
               
                 GCTGCGATGATGACAGAGGTTCTTGCACATAAGATAAGGGCAGTGGAAAAGAAACCTATTCCCCGAAT 
               
               
                   
               
               
                 CAAGCGGAAGAATAACTCAACAACAACAAACAACAGTAAGCCTTCGTCACTGCCAACCTTAGGGAGTA 
               
               
                   
               
               
                 ACACTGAGACCGTGCAACCTGAAGTAAAAAGTGAAACCGAACCCCATTTTATCTTAAAAAGTTCAGAC 
               
               
                   
               
               
                 AACACTAAAACTTATTCGCTGATGCCATCCGCTCCTCACCCAGTGAAAGAGGCATCTCCAGGCTTCTC 
               
               
                   
               
               
                 CTGGTCCCCGAAGACTGCTTCAGCCACACCAGCTCCACTGAAGAATGACGCAACAGCCTCATGCGGGT 
               
               
                   
               
               
                 TTTCAGAAAGAAGCAGCACTCCCCACTGTACGATGCCTTCGGGAAGACTCAGTGGTGCCAATGCTGCA 
               
               
                   
               
               
                 GCTGCTGATGGCCCTGGCATTTCACAGCTTGGCGAAGTGGCTCCTCTCCCCACCCTGTCTGCTCCTGT 
               
               
                   
               
               
                 GATGGAGCCCCTCATTAATTCTGAGCCTTCCACTGGTGTGACTGAGCCGCTAACGCCTCATCAGCCAA 
               
               
                   
               
               
                 ACCACCAGCCCTCCTTCCTCACCTCTCCTCAAGACCTTGCCTCTTCTCCAATGGAAGAAGATGAGCAG 
               
               
                   
               
               
                 CATTCTGAAGCAGATGAGCCTCCATCAGACGAACCCCTATCTGATGACCCCCTGTCACCTGCTGAGGA 
               
               
                   
               
               
                 GAAATTGCCCCACATTGATGAGTATTGGTCAGACAGTGAGCACATCTTTTTGGATGCAAATATTGGTG 
               
               
                   
               
               
                 GGGTGGCCATCGCACCTGCTCACGGCTCGGTTTTGATTGAGTGTGCCCGGCGAGAGCTGCACGCTACC 
               
               
                   
               
               
                 ACTCCTGTTGAGCACCCCAACCGTAATCATCCAACCCGCCTCTCCCTTGTCTTTTACCAGCACAAAAA 
               
               
                   
               
               
                 CCTAAATAAGCCCCAACATGGTTTTGAACTAAACAAGATTAAGTTTGAGGCTAAAGAAGCTAAGAATA 
               
               
                   
               
               
                 AGAAAATGAAGGCCTCAGAGCAAAAAGACCAGGCAGCTAATGAAGGTCCAGAACAGTCCTCTGAAGTA 
               
               
                   
               
               
                 AATGAATTGAACCAAATTCCTTCTCATAAAGCATTAACATTAACCCATGACAATGTTGTCACCGTGTC 
               
               
                   
               
               
                 CCCTTATGCTCTCACACACGTTGCGGGGCCCTATAACCATTGGGTC