Patent Publication Number: US-2022235361-A1

Title: RNAi Inhibition of Alpha-ENaC Expression

Description:
CROSS REFERENCE TO RELATED APPLICATIONS 
     This application is a divisional of U.S. patent application Ser. No. 17/560,475, filed Dec. 23, 2021, which is a divisional of U.S. patent application Ser. No. 16/752,325, filed Jan. 24, 2020, now U.S. patent Ser. No. 11/208,662, which is a divisional of U.S. patent application Ser. No. 15/874,396, filed Jan. 18, 2018, now U.S. patent Ser. No. 10/544,418, which is a continuation of U.S. patent application Ser. No. 15/260,824, filed on Sep. 9, 2016, now U.S. Pat. No. 9,914,927, which is a continuation of U.S. patent application Ser. No. 14/729,104, filed on Jun. 3, 2015, now U.S. Pat. No. 9,476,052, which is a divisional of U.S. patent application Ser. No. 12/683,146, filed on Jan. 6, 2010, now U.S. Pat. No. 9,074,212, which is a divisional of U.S. patent application Ser. No. 12/140,112, filed on Jun. 16, 2008, now U.S. Pat. No. 7,718,632, which claims priority to EP Application No. 07110376.6, filed Jun. 15, 2007 and EP Application No. 07114265.7, filed Aug. 13, 2007, the contents of each of which are incorporated herein by reference in their entirety. 
    
    
     TECHNICAL FIELD 
     The invention relates to the field of ENaC-mediated airway ion transport and compositions and methods for modulating alpha-ENaC expression, and more particularly to the down-regulation of alpha-ENaC by oligonucleotides via RNA interference which are administered locally to the lungs and nasal passage via inhalation/intranasal administration, or are administered systemically, e.g. by via intravenous injection. 
     BACKGROUND 
     RNA interference or “RNAi” is a term initially coined by Fire and co-workers to describe the observation that double-stranded RNA (dsRNA) can block gene expression when it is introduced into worms (Fire et al.,  Nature  391:806-811, 1998). Short dsRNA directs gene-specific, post-transcriptional silencing in many organisms, including vertebrates, and has provided a new tool for studying gene function. This technology has been reviewed numerous times recently, see, for example Novina, C. D, and Sharp, P., Nature 2004, 430:161, and Sandy, P., et al., Biotechniques 2005, 39:215, hereby incorporated by reference. 
     The mucosal surfaces at the interface between the environment and the body have evolved a number of protective mechanisms. A principal form of such innate defense is to cleanse these surfaces with liquid. Typically, the quantity of the liquid layer on a mucosal surface reflects the balance between epithelial liquid secretion, often reflecting anion (Cl −  and/or HCO 3   − ) secretion coupled with water (and a cation counter-ion), and epithelial liquid absorption, often reflecting Na +  absorption, coupled with water and counter anion (Cl −  and/or HCO 3   − ). Many diseases of mucosal surfaces are caused by too little protective liquid on those mucosal surfaces created by an imbalance between secretion (too little) and absorption (relatively too much). The defective salt transport processes that characterize these mucosal dysfunctions reside in the epithelial layer of the mucosal surface. One approach to replenish the protective liquid layer on mucosal surfaces is to “re-balance” the system by blocking Na +  channel mediated liquid absorption. The epithelial protein that mediates the rate-limiting step of Na and liquid absorption is the epithelial Na +  channel (ENaC). Alpha-ENaC is positioned on the apical surface of the epithelium, i.e. the mucosal surface-environmental interface. Inhibition of alpha-ENaC mediated Na +  mediated liquid absorption may achieve therapeutic utility. Therefore, there is a need for the development of effective therapies for the treatment and prevention of diseases or disorders in which alpha-ENaC is implicated, e.g. cystic fibrosis in humans and animals, and particularly for therapies with high efficiency. One prerequisite for high efficiency is that the active ingredient is not degraded too quickly in a physiological environment. 
     SUMMARY 
     The present invention provides specific compositions and methods that are useful in reducing alpha-ENaC levels in a subject, e.g., a mammal, such as a human, e.g. by inhaled, intranasal or intratracheal administration of such agents. 
     The present invention specifically provides iRNA agents consisting of, consisting essentially of or comprising at least 15 or more contiguous nucleotides for alpha-ENaC, and more particularly agents comprising 15 or more contiguous nucleotides from one of the sequences provided in Tables 1A-1D. The iRNA agent preferably comprises less than 30 nucleotides per strand, e.g., 21-23 nucleotides, such as those provided in Tables 1A-1D. The double stranded iRNA agent can either have blunt ends or more preferably have overhangs of 1-4 nucleotides from one or both 3′ ends of the agent. 
     Further, the iRNA agent can either contain only naturally occurring ribonucleotide subunits, or can be synthesized so as to contain one or more modifications to the sugar, phosphate or base of one or more of the ribonucleotide subunits that is included in the agent. The iRNA agent can be further modified so as to be attached to a ligand that is selected to improve stability, distribution or cellular uptake of the agent, e.g. cholesterol. The iRNA agents can further be in isolated form or can be part of a pharmaceutical composition used for the methods described herein, particularly as a pharmaceutical composition formulated for delivery to the lungs or nasal passage or formulated for parental administration. The pharmaceutical compositions can contain one or more iRNA agents, and in some embodiments, will contain two or more iRNA agents, each one directed to a different segment the alpha-ENaC gene. 
     One aspect of the present invention relates to a double-stranded oligonucleotide comprising at least one non-natural nucleobase. In certain embodiments, the non-natural nucleobase is difluorotolyl, nitroindolyl, nitropyrrolyl, or nitroimidazolyl. In a preferred embodiment, the non-natural nucleobase is difluorotolyl. In certain embodiments, only one of the two oligonucleotide strands comprising the double-stranded oligonucleotide contains a non-natural nucleobase. In certain embodiments, both of the oligonucleotide strands comprising the double-stranded oligonucleotide independently contain a non-natural nucleobase. 
     The present invention further provides methods for reducing the level of alpha-ENaC mRNA in a cell. Such methods comprise the step of administering one of the iRNA agents of the present invention to a subject as further described below. The present methods utilize the cellular mechanisms involved in RNA interference to selectively degrade the target RNA in a cell and are comprised of the step of contacting a cell with one of the iRNA agents of the present invention. Such methods can be performed directly on a cell or can be performed on a mammalian subject by administering to a subject one of the iRNA agents/pharmaceutical compositions of the present invention. Reduction of target RNA in a cell results in a reduction in the amount of encoded protein produced, and in an organism, results in reduction of epithelial potential difference, decreased fluid absorption and increased mucociliary clearance. 
     The methods and compositions of the invention, e.g., the methods and iRNA agent compositions can be used with any dosage and/or formulation described herein, as well as with any route of administration described herein. 
     The details of one or more embodiments of the invention are set forth in the accompanying drawings and the description below. Other features, objects, and advantages of the invention will be apparent from this description, the drawings, and from the claims. This application incorporates all cited references, patents, and patent applications by references in their entirety for all purposes. 
    
    
     
       BRIEF DESCRIPTION OF THE FIGURES 
         FIG. 1 : Restriction digest map of pXoon contsruct for cloned cynomolgous α-EnaC. 
         FIG. 2 : Cloning of the predicted off-target and the on-target recognition sites into the AY535007 dual luciferase reporter construct. Fragments consist of 19nt of the predicted target site and 10 nt of flanking sequence at both the 5′ and 3′ ends. 
     
    
    
     DETAILED DESCRIPTION OF CERTAIN PREFERRED EMBODIMENTS OF THE INVENTION 
     For ease of exposition the term “nucleotide” or “ribonucleotide” is sometimes used herein in reference to one or more monomeric subunits of an RNA agent. It will be understood that the usage of the term “ribonucleotide” or “nucleotide” herein can, in the case of a modified RNA or nucleotide surrogate, also refer to a modified nucleotide, or surrogate replacement moiety, as further described below, at one or more positions. 
     An “RNA agent” as used herein, is an unmodified RNA, modified RNA, or nucleoside surrogate, each of which is described herein or is well known in the RNA synthetic art. While numerous modified RNAs and nucleoside surrogates are described, preferred examples include those which have greater resistance to nuclease degradation than do unmodified RNAs. Preferred examples include those that have a 2′ sugar modification, a modification in a single strand overhang, preferably a 3′ single strand overhang, or, particularly if single stranded, a 5′-modification which includes one or more phosphate groups or one or more analogs of a phosphate group. 
     An “iRNA agent” (abbreviation for “interfering RNA agent”) as used herein, is an RNA agent, which can downregulate the expression of a target gene, e.g. ENaC gene SCNN1A. While not wishing to be bound by theory, an iRNA agent may act by one or more of a number of mechanisms, including post-transcriptional cleavage of a target mRNA sometimes referred to in the art as RNAi, or pre-transcriptional or pre-translational mechanisms. 
     A “ds iRNA agent” (abbreviation for “double stranded iRNA agent”), as used herein, is an iRNA agent which includes more than one, and preferably two, strands in which interstrand hybridization can form a region of duplex structure. A “strand” herein refers to a contigouous sequence of nucleotides (including non-naturally occurring or modified nucleotides). The two or more strands may be, or each form a part of, separate molecules, or they may be covalently interconnected, e.g., by a linker, e.g., a polyethyleneglycol linker, to form one molecule. At least one strand can include a region which is sufficiently complementary to a target RNA. Such strand is termed the “antisense strand.” A second strand of the dsRNA agent, which comprises a region complementary to the antisense strand, is termed the “sense strand.” However, a ds iRNA agent can also be formed from a single RNA molecule which is at least partly self-complementary, forming, e.g., a hairpin or panhandle structure, including a duplex region. The latter are herein referred to as short hairpin RNAs or shRNAs. In such case, the term “strand” refers to one of the regions of the RNA molecule that is complementary to another region of the same RNA molecule. 
     Although, in mammalian cells, long ds iRNA agents can induce the interferon response which is frequently deleterious, short ds iRNA agents do not trigger the interferon response, at least not to an extent that is deleterious to the cell and/or host (Manche et al.,  Mol. Cell. Biol.  12:5238, 1992; Lee et al.,  Virology  199:491, 1994; Castelli et al.,  J. Exp. Med.  186:967, 1997; Zheng et al.,  RNA  10:1934, 2004; Heidel et al.,  Nature Biotechnol.  22 1579). The iRNA agents of the present invention include molecules which are sufficiently short that they do not trigger a deleterious non-specific interferon response in normal mammalian cells. Thus, the administration of a composition including an iRNA agent (e.g., formulated as described herein) to a subject can be used to decrease expression of alpha-ENaC in the subject, while circumventing an interferon response. Molecules that are short enough that they do not trigger a deleterious interferon response are termed siRNA agents or siRNAs herein. “siRNA agent” or “siRNA” as used herein, refers to an iRNA agent, e.g., a ds iRNA agent, that is sufficiently short that it does not induce a deleterious interferon response in a mammalian, and particularly a human, cell, e.g., it has a duplexed region of less than 60 but preferably less than 50, 40, or 30 nucleotide pairs. 
     The isolated iRNA agents described herein, including ds iRNA agents and siRNA agents, can mediate the decreased expression of alpha-ENaC, e.g., by RNA degradation. For convenience, such RNA is also referred to herein as the RNA to be silenced. Such a nucleic acid is also referred to as a “target RNA”, sometimes “target RNA molecule” or sometimes “target gene”. 
     As used herein, the phrase “mediates RNAi” refers to the ability of an agent to silence, in a sequence-specific manner, a target gene. “Silencing a target gene” means the process whereby a cell containing and/or expressing a certain product of the target gene when not in contact with the agent, will contain and/or express at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% less of such gene product when contacted with the agent, as compared to a similar cell which has not been contacted with the agent. Such product of the target gene can, for example, be a messenger RNA (mRNA), a protein, or a regulatory element. 
     As used herein, the term “complementary” is used to indicate a sufficient degree of complementarity such that stable and specific binding occurs between a compound of the invention and a target RNA molecule, e.g., alpha-ENaC mRNA. Specific binding requires a sufficient degree of complementarity to avoid non-specific binding of the oligomeric compound to non-target sequences under conditions in which specific binding is desired, i.e., under physiological conditions in the case of in vivo assays or therapeutic treatment, or in the case of in vitro assays, under conditions in which the assays are performed. The non-target sequences typically differ from the target sequences by at least 2, 3 or 4 nucleotides. 
     As used herein, an iRNA agent is “sufficiently complementary” to a target RNA, e.g., a target mRNA (e.g., alpha-ENaC mRNA) if the iRNA agent reduces the production of a protein encoded by the target RNA in a cell. The iRNA agent may also be “exactly complementary” to the target RNA, e.g., the target RNA and the iRNA agent anneal, preferably to form a hybrid made exclusively of Watson-Crick basepairs in the region of exact complementarity. A “sufficiently complementary” iRNA agent can include an internal region (e.g., of at least 10 nucleotides) that is exactly complementary to a target alpha-ENaC RNA. Moreover, in some embodiments, the iRNA agent specifically discriminates a single-nucleotide difference. In this case, the iRNA agent only mediates RNAi if exact complementarity is found in the region (e.g., within 7 nucleotides of) the single-nucleotide difference. Preferred iRNA agents will be based on or consist of or comprise the sense and antisense sequences provided in Tables 1A-1D. 
     As used herein, “essentially identical” when used referring to a first nucleotide sequence in comparison to a second nucleotide sequence means that the first nucleotide sequence is identical to the second nucleotide sequence except for up to one, two or three nucleotide substitutions (e.g., adenosine replaced by uracil). “Essentially retaining the ability to inhibit alpha-ENaC expression in cultured human cells,” as used herein referring to an iRNA agent not identical to but derived from one of the iRNA agents of Tables 1A-1D by deletion, addition or substitution of nucleotides, means that the derived iRNA agent possesses an inhibitory activity not less than 20% of the inhibitory activity of the iRNA agent of Tables 1A-1D from which it was derived. For example, an iRNA agent derived from an iRNA agent of Tables 1A-1D which lowers the amount of alpha-ENaC mRNA present in cultured human cells by 70% may itself lower the amount of mRNA present in cultured human cells by at least 50% in order to be considered as essentially retaining the ability to inhibit alpha-ENaC replication in cultured human cells. Optionally, an iRNA agent of the invention may lower the amount of alpha-ENaC mRNA present in cultured human cells by at least 50%. 
     As used herein, a “subject” refers to a mammalian organism undergoing treatment for a disorder mediated by alpha-ENaC. The subject can be any mammal, such as a cow, horse, mouse, rat, dog, pig, goat, or a primate. In the preferred embodiment, the subject is a human. 
     Design and Selection of iRNA Agents 
     As used herein, “disorders associated with alpha-ENaC expression” refers to any biological or pathological state that (1) is mediated at least in part by the presence of alpha-ENaC and (2) whose outcome can be affected by reducing the level of the alpha-ENaC present. Specific disorders associated with alpha-ENaC expression are noted below. 
     The present invention is based on the design, synthesis and generation of iRNA agents that target alpha-ENaC and the demonstration of silencing of the alpha-ENaC gene in vitro in cultured cells after incubation with an iRNA agent, and the resulting protective effect towards alpha-ENaC mediated disorders. 
     An iRNA agent can be rationally designed based on sequence information and desired characteristics. For example, an iRNA agent can be designed according to the relative melting temperature of the candidate duplex. Generally, the duplex should have a lower melting temperature at the 5′ end of the antisense strand than at the 3′ end of the antisense strand. 
     The present invention provides compositions containing siRNA(s) and/or shRNA(s) targeted to one or more alpha-ENaC transcripts. 
     For any particular gene target that is selected, the design of siRNAs or shRNAs for use in accordance with the present invention will preferably follow certain guidelines. Also, in many cases, the agent that is delivered to a cell according to the present invention may undergo one or more processing steps before becoming an active suppressing agent (see below for further discussion); in such cases, those of ordinary skill in the art will appreciate that the relevant agent will preferably be designed to include sequences that may be necessary for its processing. 
     Diseases mediated by dysfunction of the epithelial sodium channel, include diseases associated with the regulation of fluid volumes across epithelial membranes. For example, the volume of airway surface liquid is a key regulator of mucociliary clearance and the maintenance of lung health. The blockade of the epithelial sodium channel will promote fluid accumulation on the mucosal side of the airway epithelium thereby promoting mucus clearance and preventing the accumulation of mucus and sputum in respiratory tissues (including lung airways). Such diseases include respiratory diseases, such as cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory tract infections (acute and chronic; viral and bacterial) and lung carcinoma. Diseases mediated by blockade of the epithelial sodium channel also include diseases other than respiratory diseases that are associated with abnormal fluid regulation across an epithelium, perhaps involving abnormal physiology of the protective surface liquids on their surface, e.g., xerostomia (dry mouth) or keratoconjunctivitis sire (dry eye). Furthermore, blockade of the epithelial sodium channel in the kidney could be used to promote diuresis and thereby induce a hypotensive effect. 
     Treatment in accordance with the invention may be symptomatic or prophylactic. 
     Asthma includes both intrinsic (non-allergic) asthma and extrinsic (allergic) asthma, mild asthma, moderate asthma, severe asthma, bronchitic asthma, exercise-induced asthma, occupational asthma and asthma induced following bacterial infection. Treatment of asthma is also to be understood as embracing treatment of subjects, e.g., of less than 4 or 5 years of age, exhibiting wheezing symptoms and diagnosed or diagnosable as “wheezy infants”, an established patient category of major medical concern and now often identified as incipient or early-phase asthmatics. (For convenience this particular asthmatic condition is referred to as “wheezy-infant syndrome”.) 
     Prophylactic efficacy in the treatment of asthma will be evidenced by reduced frequency or severity of symptomatic attack, e.g., of acute asthmatic or bronchoconstrictor attack, improvement in lung function or improved airways hyperreactivity. It may further be evidenced by reduced requirement for other, symptomatic therapy, i.e., therapy for or intended to restrict or abort symptomatic attack when it occurs, e.g., anti-inflammatory (e.g., corticosteroid) or bronchodilatory. Prophylactic benefit in asthma may, in particular, be apparent in subjects prone to “morning dipping”. “Morning dipping” is a recognized asthmatic syndrome, common to a substantial percentage of asthmatics and characterized by asthma attack, e.g., between the hours of about 4-6 am, i.e., at a time normally substantially distant from any previously administered symptomatic asthma therapy. 
     Chronic obstructive pulmonary disease includes chronic bronchitis or dyspnea associated therewith, emphysema, as well as exacerbation of airways hyperreactivity consequent to other drug therapy, in particular, other inhaled drug therapy. The invention is also applicable to the treatment of bronchitis of whatever type or genesis including, e.g., acute, arachidic, catarrhal, croupus, chronic or phthinoid bronchitis. 
     Based on the results shown herein, the present invention provides iRNA agents that reduce alpha-ENaC expression in cultured cells and in a subject, e.g. a mammalian, for example a human. Tables 1A-1D provide exemplary iRNA agents targeting alpha-ENaC, based on the standard nomenclature abbreviations given in Table A. 
     Table 1A, Seq Id No.s 305-608, Table 1B and Table 1D, Seq Id No.s 1519-1644 list siRNAs that do not comprise nucleotide modifications except for one phosphorothioate linkage between the 3′-terminal and the penultimate thymidines. The remaining Seq Ids in Tables 1A-1D lists siRNAs wherein all nucleotides comprising pyrimidine bases are 2′-O-methyl-modified nucleotides in the sense strand, and all uridines in a sequence context of 5′-ua-3′ as well as all cytidines in a sequence context of or 5′-ca-3′ are 2′-O-methyl-modified nucleotides in the antisense strand. 
     Based on these results, the invention specifically provides an iRNA agent that includes a sense strand having at least 15 contiguous nucleotides of the sense strand sequences of the agents provided in Tables 1A-1D, and an antisense strand having at least 15 contiguous nucleotides of the antisense sequences of the agents provided in Tables 1A-1D. 
     The iRNA agents shown in Tables 1A-1D are composed of two strands of 19 nucleotides in length which are complementary or identical to the target sequence, plus a 3′-TT overhang. The present invention provides agents that comprise at least 15, or at least 16, 17, or 18, or 19 contiguous nucleotides from these sequences. However, while these lengths may potentially be optimal, the iRNA agents are not meant to be limited to these lengths. The skilled person is well aware that shorter or longer iRNA agents may be similarly effective, since, within certain length ranges, the efficacy is rather a function of the nucleotide sequence than strand length. For example, Yang, et al.,  PNAS  99:9942-9947 (2002), demonstrated similar efficacies for iRNA agents of lengths between 21 and 30 base pairs. Others have shown effective silencing of genes by iRNA agents down to a length of approx. 15 base pairs (Byrom, et al., “Inducing RNAi with siRNA Cocktails Generated by RNase III”  Tech Notes  10(1), Ambion, Inc., Austin, Tex.). 
     Therefore, it is possible and contemplated by the instant invention to select from the sequences provided in Tables 1A-1D a partial sequence of between 15 to 19 nucleotides for the generation of an iRNA agent derived from one of the sequences provided in Tables 1A-1D. Alternatively, one may add one or several nucleotides to one of the sequences provided in Tables 1A-1D, or an agent comprising 15 contiguous nucleotides from one of these agents, preferably, but not necessarily, in such a fashion that the added nucleotides are complementary to the respective sequence of the target gene, e.g., alpha-ENaC. For example, the first 15 nucleotides from one of the agents can be combined with the 8 nucleotides found 5′ to these sequence in alpha-ENaC mRNA to obtain an agent with 23 nucleotides in the sense and antisense strands. All such derived iRNA agents are included in the iRNA agents of the present invention, provided they essentially retain the ability to inhibit alpha-ENaC replication in cultured human cells. 
     The antisense strand of an iRNA agent should be equal to or at least, 14, 15, 16, 17, 18, 19, 25, 29, 40, or 50 nucleotides in length. It should be equal to or less than 60, 50, 40, or 30, nucleotides in length. Preferred ranges are 15-30, 17 to 25, 19 to 23, and 19 to 21 nucleotides in length. 
     The sense strand of an iRNA agent should be equal to or at least 14, 15, 16 17, 18, 19, 25, 29, 40, or 50 nucleotides in length. It should be equal to or less than 60, 50, 40, or 30 nucleotides in length. Preferred ranges are 15-30, 17 to 25, 19 to 23, and 19 to 21 nucleotides in length. 
     The double stranded portion of an iRNA agent should be equal to or at least, 15, 16 17, 18, 19, 20, 21, 22, 23, 24, 25, 29, 40, or 50 nucleotide pairs in length. It should be equal to or less than 60, 50, 40, or 30 nucleotides pairs in length. Preferred ranges are 15-30, 17 to 25, 19 to 23, and 19 to 21 nucleotides pairs in length. 
     Generally, the iRNA agents of the instant invention include a region of sufficient complementarity to the alpha-ENaC mRNA, and are of sufficient length in terms of nucleotides, that the iRNA agent, or a fragment thereof, can mediate down regulation of the alpha-ENaC gene. It is not necessary that there be perfect complementarity between the iRNA agent and the target gene, but the correspondence must be sufficient to enable the iRNA agent, or a cleavage product thereof, to direct sequence specific silencing, e.g., by RNAi cleavage of an alpha-ENaC mRNA. 
     Therefore, the iRNA agents of the instant invention include agents comprising a sense strand and antisense strand each comprising a sequence of at least 16, 17 or 18 nucleotides which is essentially identical, as defined below, to one of the sequences of Tables 1A-1D, except that not more than 1, 2 or 3 nucleotides per strand, respectively, have been substituted by other nucleotides (e.g. adenosine replaced by uracil), while essentially retaining the ability to inhibit alpha-ENaC expression in cultured human cells. These agents will therefore possess at least 15 nucleotides identical to one of the sequences of Tables 1A-1D, but 1, 2 or 3 base mismatches with respect to either the target alpha-ENaC sequence or between the sense and antisense strand are introduced. Mismatches to the target alpha-ENaC RNA sequence, particularly in the antisense strand, are most tolerated in the terminal regions and if present are preferably in a terminal region or regions, e.g., within 6, 5, 4, or 3 nucleotides of a 5′ and/or 3′ terminus, most preferably within 6, 5, 4, or 3 nucleotides of the 5′-terminus of the sense strand or the 3′-terminus of the antisense strand. The sense strand need only be sufficiently complementary with the antisense strand to maintain the overall double stranded character of the molecule. 
     It is preferred that the sense and antisense strands be chosen such that the iRNA agent includes a single strand or unpaired region at one or both ends of the molecule. Thus, an iRNA agent contains sense and antisense strands, preferably paired to contain an overhang, e.g., one or two 5′ or 3′ overhangs but preferably a 3′ overhang of 2-3 nucleotides. Most embodiments will have a 3′ overhang. Preferred siRNA agents will have single-stranded overhangs, preferably 3′ overhangs, of 1 to 4, or preferably 2 or 3 nucleotides, in length, at one or both ends of the iRNA agent. The overhangs can be the result of one strand being longer than the other, or the result of two strands of the same length being staggered. The unpaired nucleotides forming the overhang can be ribonucleotides, or they can be deoxyribonucleotides, preferably thymidine. 5′-ends are preferably phosphorylated, or they may be unphosphorylated. 
     Preferred lengths for the duplexed region are between 15 and 30, most preferably 18, 19, 20, 21, 22, and 23 nucleotides in length, e.g., in the siRNA agent range discussed above. siRNA agents can resemble in length and structure the natural Dicer processed products from long dsRNAs. Embodiments in which the two strands of the siRNA agent are linked, e.g., covalently linked, are also included. Hairpin, or other single strand structures which provide the required double stranded region, and preferably a 3′ overhang are also within the invention. 
     Evaluation of Candidate iRNA Agents 
     As noted above, the present invention provides a system for identifying siRNAs that are useful as inhibitors of alpha-ENaC. Since, as noted above, shRNAs are processed intracellularly to produce siRNAs having duplex portions with the same sequence as the stem structure of the shRNA, the system is equally useful for identifying shRNAs that are useful as inhibitors of alpha-ENaC. For purposes of description this section will refer to siRNAs, but the system also encompasses corresponding shRNAs. Specifically, the present invention demonstrates the successful preparation of siRNAs targeted to inhibit alpha-ENaC activity. The techniques and reagents described herein can readily be applied to design potential new siRNAs, targeted to other genes or gene regions, and tested for their activity in inhibiting alpha-ENaC as discussed herein. 
     In various embodiments of the invention potential alpha-ENaC inhibitors can be tested for suppression of endogenous alpha-ENaC expression by introducing candidate siRNA(s) into cells (e.g., by exogenous administration or by introducing a vector or construct that directs endogenous synthesis of siRNA into the cell), or in laboratory animals by pulmonary or nasal administration. Alternately, potential alpha-ENaC inhibitors can be tested in vitro by transient co-transfection of candidate siRNA(s) together with an alpha-ENaC-expression plasmid. The ability of the candidate siRNA(s) to reduce target transcript levels and/or to inhibit or suppress one or more aspects or features of alpha-ENaC activity such as epithelial potential difference or airway surface fluid absorption is then assessed. 
     Cells or laboratory animals to which inventive siRNA compositions have been delivered (test cells/animals) may be compared with similar or comparable cells or laboratory animals that have not received the inventive composition (control cells/animals, e.g., cells/animals that have received either no siRNA or a control siRNA such as an siRNA targeted to a non-endogenous transcript such as green fluorescent protein (GFP)). The ion transport phenotype of the test cells/animals can be compared with the phenotype of control cells/animals, providing that the inventive siRNA share sequence cross-reactivity with the test cell type/species. Production of alpha-ENaC protein and short circuit current (in vitro or ex vivo) may be compared in the test cells/animals relative to the control cells/animals. Other indicia of alpha-ENaC activity, including ex vivo epithelial potential difference or in vivo mucocilliary clearance or whole body magnetic resonance imaging (MRI), can be similarly compared. Generally, test cells/animals and control cells/animals would be from the same species and, for cells, of similar or identical cell type. For example, cells from the same cell line could be compared. When the test cell is a primary cell, typically the control cell would also be a primary cell. 
     For example, the ability of a candidate siRNA to inhibit alpha-ENaC activity may conveniently be determined by (i) delivering the candidate siRNA to cells (ii) assessing the expression levels of alpha-ENaC mRNA relative to an endogenously expressed control gene (iii) comparing the amiloride-sensitive current in an in vitro cell model produced in the presence of the siRNA with the amount produced in the absence of the siRNA. This latter assay may be used to test siRNAs that target any target transcript that may influence alpha-ENaC activity indirectly and is not limited to siRNAs that target the transcripts that encode the ENaC channel subunits. 
     The ability of a candidate siRNA to reduce the level of the target transcript may be assessed by measuring the amount of the target transcript using, for example, Northern blots, nuclease protection assays, probe hybridization, reverse transcription (RT)-PCR, real-time RT-PCR, microarray analysis, etc. The ability of a candidate siRNA to inhibit production of a polypeptide encoded by the target transcript (either at the transcriptional or post-transcriptional level) may be measured using a variety of antibody-based approaches including, but not limited to, Western blots, immunoassays, ELISA, flow cytometry, protein microarrays, etc. In general, any method of measuring the amount of either the target transcript or a polypeptide encoded by the target transcript may be used. 
     In general, certain preferred alpha-ENaC iRNA inhibitors reduce the target transcript level at least about 2 fold, preferably at least about 4 fold, more preferably at least about 8 fold, at least about 16 fold, at least about 64 fold or to an even greater degree relative to the level that would be present in the absence of the inhibitor (e.g., in a comparable control cell lacking the inhibitor). In general, certain preferred alpha-ENaC iRNA inhibitors inhibit ENaC channel activity, so that the activity is lower in a cell containing the inhibitor than in a control cell not containing the inhibitor by at least about 2 fold, preferably at least about 4 fold, more preferably at least about 8 fold, at least about 16 fold, at least about 64 fold, at least about 100 fold, at least about 200 fold, or to an even greater degree. 
     Certain preferred alpha-ENaC iRNA inhibitors inhibit ENaC channel activity for at least 12 hours, at least 24 hours, at least 36 hours, at least 48 hours, at least 60 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 144 hours or at least 168 hours following administration of the siRNA and infection of the cells. Certain preferred alpha-ENaC inhibitors prevent (i.e., reduce to undetectable levels) or significantly reduce alpha-ENaC activity for at least 24 hours, at least 36 hours, at least 48 hours, or at least 60 hours following administration of the siRNA. According to various embodiments of the invention a significant reduction in alpha-ENaC activity is a reduction to less than approximately 90% of the level that would occur in the absence of the siRNA, a reduction to less than approximately 75% of the level that would occur in the absence of the siRNA, a reduction to less than approximately 50% of the level that would occur in the absence of the siRNA, a reduction to less than approximately 25% of the level that would occur in the absence of the siRNA, or a reduction to less than approximately 10% of the level that would occur in the absence of the siRNA. Reduction in alpha-ENaC activity may be measured using any suitable method including, but not limited to, short circuit current measurement of amiloride sensitivity in vitro, epithelial potential difference ex vivo or in vivo mucocilliary clearance or whole body/lung MRI. 
     Stability Testing, Modification, and Retesting of iRNA Agents 
     A candidate iRNA agent can be evaluated with respect to stability, e.g., its susceptibility to cleavage by an endonuclease or exonuclease, such as when the iRNA agent is introduced into the body of a subject. Methods can be employed to identify sites that are susceptible to modification, particularly cleavage, e.g., cleavage by a component found in the body of a subject. Such methods may include the isolation and identification of most abundant fragments formed by degradation of the candidate iRNA agent after its incubation with isolated biological media in vitro, e.g. serum, plasma, sputum, cerebrospinal fluid, or cell or tissue homogenates, or after contacting a subject with the candidate iRNA agent in vivo, thereby identifying sites prone to cleavage. Such methods are, for example, without limitation, in International Patent Application Publication No. WO2005115481, filed on May 27, 2005. 
     When sites susceptible to cleavage are identified, a further iRNA agent can be designed and/or synthesized wherein the potential cleavage site is made resistant to cleavage, e.g. by introduction of a 2′-modification on the site of cleavage, e.g. a 2′-O-methyl group. This further iRNA agent can be retested for stability, and this process may be iterated until an iRNA agent is found exhibiting the desired stability. 
     In Vivo Testing 
     An iRNA agent identified as being capable of inhibiting alpha-ENaC gene expression can be tested for functionality in vivo in an animal model (e.g., in a mammal, such as in mouse, rat, guinea-pig or primate). For example, the iRNA agent can be administered to an animal, and the iRNA agent evaluated with respect to its biodistribution, stability, and its ability to inhibit alpha-ENaC expression or modulate a biological or pathological process mediated at least in part by alpha-ENaC. 
     The iRNA agent can be administered directly to the target tissue, such as by injection, or the iRNA agent can be administered to the animal model in the same manner that it would be administered to a human. Preferably, the iRNA agent is delivered to the subject&#39;s airways, such as by intranasal, inhaled or intratracheal administration. 
     The iRNA agent can also be evaluated for its intracellular distribution. The evaluation can include determining whether the iRNA agent was taken up into the cell. The evaluation can also include determining the stability (e.g., the half-life) of the iRNA agent. Evaluation of an iRNA agent in vivo can be facilitated by use of an iRNA agent conjugated to a traceable marker (e.g., a fluorescent marker such as fluorescein; a radioactive label, such as  35 S,  32 P,  33 P, or  3 H; gold particles; or antigen particles for immunohistochemistry). 
     The iRNA agent can be evaluated with respect to its ability to down regulate alpha-alpha-ENaC expression. Levels of alpha-ENaC gene expression in vivo can be measured, for example, by in situ hybridization, or by the isolation of RNA from tissue prior to and following exposure to the iRNA agent. Where the animal needs to be sacrificed in order to harvest the tissue, an untreated control animal will serve for comparison. alpha-ENaC RNA can be detected by any desired method, including but not limited to RT-PCR, northern blot, branched-DNA assay, or RNAase protection assay. Alternatively, or additionally, alpha-ENaC gene expression can be monitored by performing western blot analysis or immunostaining on tissue extracts treated with the iRNA agent. 
     Potential alpha-ENaC inhibitors can be tested using any variety of animal models that have been developed. Compositions comprising candidate siRNA(s), constructs or vectors capable of directing synthesis of such siRNAs within a host cell, or cells engineered or manipulated to contain candidate siRNAs may be administered to an animal. The ability of the composition to suppress alpha-ENaC expression and/or to modify ENaC-dependent phenotypes and/or lessen their severity relative to animals that have not received the potential alpha-ENaC inhibitor is assessed. Such models include, but are not limited to, murine, rat, guinea pig, sheep and non-human primate models for ENaC-dependent phenotypes, all of which are known in the art and are used for testing the efficacy of potential alpha-ENaC therapeutics. 
     Utilising the systems invented for identifying candidate therapeutic siRNA agents, suitable therapeutic agents are selected from Duplex identifiers ND-8302, ND-8332, ND-8348, ND-8356, ND-8357, ND-8373, ND-8381, ND-8396, ND-8450 and ND-8453, more suitably selected from ND-8356, ND-8357 and ND-8396. 
     iRNA Chemistry Described herein are isolated iRNA agents, e.g., ds RNA agents that mediate RNAi to inhibit expression of the alpha-ENaC gene. 
     RNA agents discussed herein include otherwise unmodified RNA as well as RNA which has been modified, e.g., to improve efficacy, and polymers of nucleoside surrogates. Unmodified RNA refers to a molecule in which the components of the nucleic acid, namely sugars, bases, and phosphate moieties, are the same or essentially the same as that which occur in nature, preferably as occur naturally in the human body. The art has referred to rare or unusual, but naturally occurring, RNAs as modified RNAs, see, e.g., Limbach et al.  Nucleic Acids Res.  22: 2183-2196, 1994. Such rare or unusual RNAs, often termed modified RNAs (apparently because they are typically the result of a post-transcriptional modification) are within the term unmodified RNA, as used herein. Modified RNA as used herein refers to a molecule in which one or more of the components of the nucleic acid, namely sugars, bases, and phosphate moieties, are different from that which occurs in nature, preferably different from that which occurs in the human body. While they are referred to as modified “RNAs,” they will of course, because of the modification, include molecules which are not RNAs. Nucleoside surrogates are molecules in which the ribophosphate backbone is replaced with a non-ribophosphate construct that allows the bases to the presented in the correct spatial relationship such that hybridization is substantially similar to what is seen with a ribophosphate backbone, e.g., non-charged mimics of the ribophosphate backbone. Examples of the above are discussed herein. 
     Modifications described herein can be incorporated into any double-stranded RNA and RNA-like molecule described herein, e.g., an iRNA agent. It may be desirable to modify one or both of the antisense and sense strands of an iRNA agent. As nucleic acids are polymers of subunits or monomers, many of the modifications described below occur at a position which is repeated within a nucleic acid, e.g., a modification of a base, or a phosphate moiety, or the non-linking oxygen of a phosphate moiety. In some cases the modification will occur at all of the subject positions in the nucleic acid but in many, and in fact in most, cases it will not. By way of example, a modification may only occur at a 3′ or 5′ terminal position, may only occur in a terminal region, e.g. at a position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10 nucleotides of a strand. A modification may occur in a double strand region, a single strand region, or in both. E.g., a phosphorothioate modification at a non-linking 0 position may only occur at one or both termini, may only occur in a terminal regions, e.g., at a position on a terminal nucleotide or in the last 2, 3, 4, 5, or 10 nucleotides of a strand, or may occur in double strand and single strand regions, particularly at termini. Similarly, a modification may occur on the sense strand, antisense strand, or both. In some cases, the sense and antisense strand will have the same modifications or the same class of modifications, but in other cases the sense and antisense strand will have different modifications, e.g., in some cases it may be desirable to modify only one strand, e.g. the sense strand. 
     Two prime objectives for the introduction of modifications into iRNA agents is their stabilization towards degradation in biological environments and the improvement of pharmacological properties, e.g. pharmacodynamic properties, which are further discussed below. Other suitable modifications to a sugar, base, or backbone of an iRNA agent are described in PCT Application No. PCT/US2004/01193, filed Jan. 16, 2004. An iRNA agent can include a non-naturally occurring base, such as the bases described in PCT Application No. PCT/US2004/011822, filed Apr. 16, 2004. An iRNA agent can include a non-naturally occurring sugar, such as a non-carbohydrate cyclic carrier molecule. Exemplary features of non-naturally occurring sugars for use in iRNA agents are described in PCT Application No. PCT/US2004/11829, filed Apr. 16, 2003. 
     An iRNA agent can include an internucleotide linkage (e.g., the chiral phosphorothioate linkage) useful for increasing nuclease resistance. In addition, or in the alternative, an iRNA agent can include a ribose mimic for increased nuclease resistance. Exemplary internucleotide linkages and ribose mimics for increased nuclease resistance are described in PCT Application No. PCT/US2004/07070, filed on Mar. 8, 2004. 
     An iRNA agent can include ligand-conjugated monomer subunits and monomers for oligonucleotide synthesis. Exemplary monomers are described in U.S. application Ser. No. 10/916,185, filed on Aug. 10, 2004. 
     An iRNA agent can have a ZXY structure, such as is described in PCT Application No. PCT/US2004/07070, filed on Mar. 8, 2004. 
     An iRNA agent can be complexed with an amphipathic moiety. Exemplary amphipathic moieties for use with iRNA agents are described in PCT Application No. PCT/US2004/07070, filed on Mar. 8, 2004. 
     In another embodiment, the iRNA agent can be complexed to a delivery agent that features a modular complex. The complex can include a carrier agent linked to one or more of (preferably two or more, more preferably all three of): (a) a condensing agent (e.g., an agent capable of attracting, e.g., binding, a nucleic acid, e.g., through ionic or electrostatic interactions); (b) a fusogenic agent (e.g., an agent capable of fusing and/or being transported through a cell membrane); and (c) a targeting group, e.g., a cell or tissue targeting agent, e.g., a lectin, glycoprotein, lipid or protein, e.g., an antibody, that binds to a specified cell type. iRNA agents complexed to a delivery agent are described in PCT Application No. PCT/US2004/07070, filed on Mar. 8, 2004. 
     An iRNA agent can have non-canonical pairings, such as between the sense and antisense sequences of the iRNA duplex. Exemplary features of non-canonical iRNA agents are described in PCT Application No. PCT/US2004/07070, filed on Mar. 8, 2004. 
     Enhanced Nuclease Resistance 
     An iRNA agent, e.g., an iRNA agent that targets alpha-ENaC, can have enhanced resistance to nucleases. 
     One way to increase resistance is to identify cleavage sites and modify such sites to inhibit cleavage, as described in U.S. Application No. 60/559,917, filed on May 4, 2004. For example, the dinucleotides 5′-ua-3′, 5′-ca-3′, 5′-ug-3′, 5′-uu-3′, or 5′-cc-3′ can serve as cleavage sites. In certain embodiments, all the pyrimidines of an iRNA agent carry a 2′-modification in either the sense strand, the antisense strand, or both strands, and the iRNA agent therefore has enhanced resistance to endonucleases. Enhanced nuclease resistance can also be achieved by modifying the 5′ nucleotide, resulting, for example, in at least one 5′-uridine-adenine-3′ (5′-ua-3′) dinucleotide wherein the uridine is a 2′-modified nucleotide; at least one 5′-cytidine-adenine-3′ (5′-ca-3′) dinucleotide, wherein the 5′-cytidine is a 2′-modified nucleotide; at least one 5′-uridine-guanine-3′ (5′-ug-3′) dinucleotide, wherein the 5′-uridine is a 2′-modified nucleotide; at least one 5′-uridine-uridine-3′ (5′-uu-3′) dinucleotide, wherein the 5′-uridine is a 2′-modified nucleotide; or at least one 5′-cytidine-cytidine-3′ (5′-cc-3′) dinucleotide, wherein the 5′-cytidine is a 2′-modified nucleotide, as described in International Application No. PCT/US2005/018931, filed on May 27, 2005. The iRNA agent can include at least 2, at least 3, at least 4 or at least 5 of such dinucleotides. In a particularly preferred embodiment, the 5′ nucleotide in all occurrences of the sequence motifs 5′-ua-3′ and 5′-ca-3′ in either the sense strand, the antisense strand, or both strands is a modified nucleotide. Preferably, the 5′ nucleotide in all occurrences of the sequence motifs 5′-ua-3′, 5′-ca-3′ and 5′-ug-3′ in either the sense strand, the antisense strand, or both strands is a modified nucleotide. More preferably, all pyrimidine nucleotides in the sense strand are modified nucleotides, and the 5′ nucleotide in all occurrences of the sequence motifs 5′-ua-3′ and 5′-ca-3′ in the antisense strand are modified nucleotides, or where the antisense strand does comprise neither of a 5′-ua-3′ and a 5′-ca-3′ motif, in all occurrences of the sequence motif 5′-ug-3′. 
     Preferably, the 2′-modified nucleotides include, for example, a 2′-modified ribose unit, e.g., the 2′-hydroxyl group (OH) can be modified or replaced with a number of different “oxy” or “deoxy” substituents. 
     Examples of “oxy”-2′ hydroxyl group modifications include alkoxy or aryloxy (OR, e.g., R=H, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl or sugar); polyethyleneglycols (PEG), O(CH 2 CH 2 O) n CH 2 CH 2 OR; “locked” nucleic acids (LNA) in which the 2′ hydroxyl is connected, e.g., by a methylene bridge, to the 4′ carbon of the same ribose sugar; O-AMINE and aminoalkoxy, O(CH 2 ) n AMINE, (e.g., AMINE=NH 2 ; alkylamino, dialkylamino, heterocyclyl amino, arylamino, diaryl amino, heteroaryl amino, or diheteroaryl amino, ethylene diamine, polyamino). It is noteworthy that oligonucleotides containing only the methoxyethyl group (MOE), (OCH 2 CH 2 OCH 3 , a PEG derivative), exhibit nuclease stabilities comparable to those modified with the robust phosphorothioate modification. 
     “Deoxy” modifications include hydrogen (i.e. deoxyribose sugars, which are of particular relevance to the overhang portions of partially ds RNA); halo (e.g., fluoro); amino (e.g. NH 2 ; alkylamino, dialkylamino, heterocyclyl, arylamino, diaryl amino, heteroaryl amino, diheteroaryl amino, or amino acid); NH(CH 2 CH 2 NH) n CH 2 CH 2 -AMINE (AMINE=NH 2 ; alkylamino, dialkylamino, heterocyclyl amino, arylamino, diaryl amino, heteroaryl amino, or diheteroaryl amino), —NHC(O)R (R=alkyl, cycloalkyl, aryl, aralkyl, heteroaryl or sugar), cyano; mercapto; alkyl-thio-alkyl; thioalkoxy; and alkyl, cycloalkyl, aryl, alkenyl and alkynyl, which may be optionally substituted with e.g., an amino functionality. 
     Preferred substitutents are 2′-methoxyethyl, 2′-OCH 3 , 2′-O-allyl, 2′-C-allyl, and 2′-fluoro. 
     The inclusion of furanose sugars in the oligonucleotide backbone can also decrease endonucleolytic cleavage. An iRNA agent can be further modified by including a 3′ cationic group, or by inverting the nucleoside at the 3′-terminus with a 3′-3′ linkage. In another alternative, the 3′-terminus can be blocked with an aminoalkyl group, e.g., a 3′ C5-aminoalkyl dT. Other 3′ conjugates can inhibit 3′-5′ exonucleolytic cleavage. While not being bound by theory, a 3′ conjugate, such as naproxen or ibuprofen, may inhibit exonucleolytic cleavage by sterically blocking the exonuclease from binding to the 3′-end of oligonucleotide. Even small alkyl chains, aryl groups, or heterocyclic conjugates or modified sugars (D-ribose, deoxyribose, glucose etc.) can block 3′-5′-exonucleases. 
     Nucleolytic cleavage can also be inhibited by the introduction of phosphate linker modifications, e.g., phosphorothioate linkages. Thus, preferred iRNA agents include nucleotide dimers enriched or pure for a particular chiral form of a modified phosphate group containing a heteroatom at a nonbridging position normally occupied by oxygen. The heteroatom can be S, Se, Nr 2 , or Br 3 . When the heteroatom is S, enriched or chirally pure Sp linkage is preferred. Enriched means at least 70, 80, 90, 95, or 99% of the preferred form. Modified phosphate linkages are particularly efficient in inhibiting exonucleolytic cleavage when introduced near the 5′- or 3′-terminal positions, and preferably the 5′-terminal positions, of an iRNA agent. 
     5′ conjugates can also inhibit 5′-3′ exonucleolytic cleavage. While not being bound by theory, a 5′ conjugate, such as naproxen or ibuprofen, may inhibit exonucleolytic cleavage by sterically blocking the exonuclease from binding to the 5′-end of oligonucleotide. Even small alkyl chains, aryl groups, or heterocyclic conjugates or modified sugars (D-ribose, deoxyribose, glucose etc.) can block 3′-5′-exonucleases. 
     An iRNA agent can have increased resistance to nucleases when a duplexed iRNA agent includes a single-stranded nucleotide overhang on at least one end. In preferred embodiments, the nucleotide overhang includes 1 to 4, preferably 2 to 3, unpaired nucleotides. In a preferred embodiment, the unpaired nucleotide of the single-stranded overhang that is directly adjacent to the terminal nucleotide pair contains a purine base, and the terminal nucleotide pair is a G-C pair, or at least two of the last four complementary nucleotide pairs are G-C pairs. In further embodiments, the nucleotide overhang may have 1 or 2 unpaired nucleotides, and in an exemplary embodiment the nucleotide overhang is 5′-gc-3′. In preferred embodiments, the nucleotide overhang is on the 3′-end of the antisense strand. In one embodiment, the iRNA agent includes the motif 5′-cgc-3′ on the 3′-end of the antisense strand, such that a 2-nt overhang 5′-gc-3′ is formed. 
     Thus, an iRNA agent can include modifications so as to inhibit degradation, e.g., by nucleases, e.g., endonucleases or exonucleases, found in the body of a subject. These monomers are referred to herein as NRMs, or Nuclease Resistance promoting Monomers, the corresponding modifications as NRM modifications. In many cases these modifications will modulate other properties of the iRNA agent as well, e.g., the ability to interact with a protein, e.g., a transport protein, e.g., serum albumin, or a member of the RISC, or the ability of the first and second sequences to form a duplex with one another or to form a duplex with another sequence, e.g., a target molecule. 
     One or more different NRM modifications can be introduced into an iRNA agent or into a sequence of an iRNA agent. An NRM modification can be used more than once in a sequence or in an iRNA agent. 
     NRM modifications include some which can be placed only at the terminus and others which can go at any position. Some NRM modifications can inhibit hybridization so it is preferable to use them only in terminal regions, and preferable to not use them at the cleavage site or in the cleavage region of a sequence which targets a subject sequence or gene, particularly on the antisense strand. They can be used anywhere in a sense strand, provided that sufficient hybridization between the two strands of the ds iRNA agent is maintained. In some embodiments it is desirable to put the NRM at the cleavage site or in the cleavage region of a sense strand, as it can minimize off-target silencing. 
     In most cases, NRM modifications will be distributed differently depending on whether they are comprised on a sense or antisense strand. If on an antisense strand, modifications which interfere with or inhibit endonuclease cleavage should not be inserted in the region which is subject to RISC mediated cleavage, e.g., the cleavage site or the cleavage region (As described in Elbashir et al., 2001, Genes and Dev. 15: 188, hereby incorporated by reference). Cleavage of the target occurs about in the middle of a 20 or 21 nt antisense strand, or about 10 or 11 nucleotides upstream of the first nucleotide on the target mRNA which is complementary to the antisense strand. As used herein cleavage site refers to the nucleotides on either side of the cleavage site, on the target or on the iRNA agent strand which hybridizes to it. Cleavage region means the nucleotides within 1, 2, or 3 nucleotides of the cleavage site, in either direction. 
     Such modifications can be introduced into the terminal regions, e.g., at the terminal position or with 2, 3, 4, or 5 positions of the terminus, of a sense or antisense strand. 
     Tethered Ligands 
     The properties of an iRNA agent, including its pharmacological properties, can be influenced and tailored, for example, by the introduction of ligands, e.g. tethered ligands. In addition, pharmacological properties of an iRNA agent can be improved by incorporating a ligand in a formulation of the iRNA agent when the iRNA agent either has or does have a tethered ligand. 
     A wide variety of entities, e.g., ligands, can be tethered to an iRNA agent or used as formulation conjugate or additive, e.g., to the carrier of a ligand-conjugated monomer subunit. Examples are described below in the context of a ligand-conjugated monomer subunit but that is only preferred, entities can be coupled at other points to an iRNA agent. 
     Preferred moieties are ligands, which are coupled, preferably covalently, either directly or indirectly, via an intervening tether to the carrier. In preferred embodiments, the ligand is attached to the carrier via an intervening tether. The ligand or tethered ligand may be present on the ligand-conjugated monomer when the ligand-conjugated monomer is incorporated into the growing strand. In some embodiments, the ligand may be incorporated into a “precursor” ligand-conjugated monomer subunit after a “precursor” ligand-conjugated monomer subunit has been incorporated into the growing strand. For example, a monomer having, e.g., an amino-terminated tether, e.g., TAP-(CH 2 ) n NH 2  may be incorporated into a growing sense or antisense strand. In a subsequent operation, i.e., after incorporation of the precursor monomer subunit into the strand, a ligand having an electrophilic group, e.g., a pentafluorophenyl ester or aldehyde group, can subsequently be attached to the precursor ligand-conjugated monomer by coupling the electrophilic group of the ligand with the terminal nucleophilic group of the precursor ligand-conjugated monomer subunit tether. 
     In preferred embodiments, a ligand alters the distribution, targeting or lifetime of an iRNA agent into which it is incorporated. In preferred embodiments a ligand provides an enhanced affinity for a selected target, e.g., molecule, cell or cell type, compartment, e.g., a cellular or organ compartment, tissue, organ or region of the body, as, e.g., compared to a species absent such a ligand. 
     Preferred ligands can improve transport, hybridization, and specificity properties and may also improve nuclease resistance of the resultant natural or modified oligoribonucleotide, or a polymeric molecule comprising any combination of monomers described herein and/or natural or modified ribonucleotides. 
     Ligands in general can include therapeutic modifiers, e.g., for enhancing uptake; diagnostic compounds or reporter groups e.g., for monitoring distribution; cross-linking agents; nuclease-resistance conferring moieties; and natural or unusual nucleobases. General examples include lipophilic molecules, lipids, lectins, steroids (e.g., uvaol, hecigenin, diosgenin), terpenes (e.g., triterpenes, e.g., sarsasapogenin, Friedelin, epifriedelanol derivatized lithocholic acid), vitamins, carbohydrates (e.g., a dextran, pullulan, chitin, chitosan, synthetic (eg Oligo Lactate 15-mer) and natural (eg low and medium molecular weight) polymers, inulin, cyclodextrin or hyaluronic acid), proteins, protein binding agents, integrin targeting molecules, polycationics, peptides, polyamines, and peptide mimics. Other examples include folic acid or epithelial cell receptor ligands, such as transferin. 
     The ligand may be a naturally occurring or recombinant or synthetic molecule, such as a synthetic polymer, e.g., a synthetic polyamino acid. Examples of polyamino acids include polylysine (PLL), poly L-aspartic acid, poly L-glutamic acid, styrene-maleic acid anhydride copolymer, poly(L-lactide-co-glycolied) copolymer, divinyl ether-maleic anhydride copolymer, N-(2-hydroxypropyl)methacrylamide copolymer (HMPA), polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyurethane, poly(2-ethylacrylic acid), N-isopropylacrylamide polymers, or polyphosphazine. Example of polyamines include: polyethylenimine, polylysine (PLL), spermine, spermidine, polyamine, pseudopeptide-polyamine, peptidomimetic polyamine, dendrimer polyamine, arginine, amidine, protamine, cationic moieties, e.g., cationic lipid, cationic porphyrin, quaternary salt of a polyamine, or an alpha helical peptide. 
     Ligands can also include targeting groups, e.g., a cell or tissue targeting agent, e.g., a thyrotropin, melanotropin, surfactant protein A, mucin carbohydrate, a glycosylated polyaminoacid, transferrin, bisphosphonate, polyglutamate, polyaspartate, or an Arg-Gly-Asp (RGD) peptide or RGD peptide mimetic. 
     Ligands can be proteins, e.g., glycoproteins, lipoproteins, e.g. low density lipoprotein (LDL), or albumins, e.g. human serum albumin (HSA), or peptides, e.g., molecules having a specific affinity for a co-ligand, or antibodies e.g., an antibody, that binds to a specified cell type such as a cancer cell, endothelial cell, or bone cell. Ligands may also include hormones and hormone receptors. They can also include non-peptidic species, such as cofactors, multivalent lactose, multivalent galactose, N-acetyl-galactosamine, N-acetyl-glucosamine, multivalent mannose, or multivalent fucose. 
     The ligand can be a substance, e.g, a drug, which can increase the uptake of the iRNA agent into the cell, for example, by disrupting the cell&#39;s cytoskeleton, e.g., by disrupting the cell&#39;s microtubules, microfilaments, and/or intermediate filaments. The drug can be, for example, taxon, vincristine, vinblastine, cytochalasin, nocodazole, japlakinolide, latrunculin A, phalloidin, swinholide A, indanocine, myoservin, tetracyclin. 
     In one aspect, the ligand is a lipid or lipid-based molecule. Such a lipid or lipid-based molecule preferably binds a serum protein, e.g., human serum albumin (HSA). An HSA binding ligand allows for distribution of the conjugate to a target tissue, e.g., liver tissue, including parenchymal cells of the liver. Other molecules that can bind HSA can also be used as ligands. For example, neproxin or aspirin can be used. A lipid or lipid-based ligand can (a) increase resistance to degradation of the conjugate, (b) increase targeting or transport into a target cell or cell membrane, and/or (c) can be used to adjust binding to a serum protein, e.g., HSA. 
     A lipid based ligand can be used to modulate, e.g., control the binding of the conjugate to a target tissue. For example, a lipid or lipid-based ligand that binds to HSA more strongly will be less likely to be targeted to the kidney and therefore less likely to be cleared from the body. 
     In a preferred embodiment, the lipid based ligand binds HSA. Preferably, it binds HSA with a sufficient affinity such that the conjugate will be preferably distributed to a non-kidney tissue. However, it is preferred that the affinity not be so strong that the HSA-ligand binding cannot be reversed. 
     In another aspect, the ligand is a moiety, e.g., a vitamin or nutrient, which is taken up by a target cell, e.g., a proliferating cell. These are particularly useful for treating disorders characterized by unwanted cell proliferation, e.g., of the malignant or non-malignant type, e.g., cancer cells. Exemplary vitamins include vitamin A, E, and K. Other exemplary vitamins include the B vitamins, e.g., folic acid, B12, riboflavin, biotin, pyridoxal or other vitamins or nutrients taken up by cancer cells. 
     In another aspect, the ligand is a cell-permeation agent, preferably a helical cell-permeation agent. Preferably, the agent is amphipathic. An exemplary agent is a peptide such as tat or antennapedia. If the agent is a peptide, it can be modified, including a peptidylmimetic, invertomers, non-peptide or pseudo-peptide linkages, and use of D-amino acids. The helical agent is preferably an alpha-helical agent, which preferably has a lipophilic and a lipophobic phase. The cell permeation agent can be linked covalently to the iRNA agent or be part of an iRNA-peptide complex. 
     5′-Phosphate Modifications 
     In preferred embodiments, iRNA agents are 5′ phosphorylated or include a phosphoryl analog at the 5′ prime terminus. 5′-phosphate modifications of the antisense strand include those which are compatible with RISC-mediated gene silencing. Suitable modifications include: 5′-monophosphate ((HO)2(O)P—O-5′); 5′-diphosphate ((HO)2(O)P—O—P(HO)(O)—O-5′); 5′-triphosphate ((HO)2(O)P—O—(HO)(O)P—O—P(HO)(O)—O-5′); 5′-guanosine cap (7-methylated or non-methylated) (7m-G-O-5′-(HO)(O)P—O—(HO)(O)P—O—P(HO)(O)—O-5′); 5′-adenosine cap (Appp), and any modified or unmodified nucleotide cap structure (N—O-5′-(HO)(O)P—O—(HO)(O)P—O—P(HO)(O)—O-5′); 5′-monothiophosphate (phosphorothioate; (HO)2(S)P—O-5′); 5′-monodithiophosphate (phosphorodithioate; (HO)(HS)(S)P—O-5′), 5′-phosphorothiolate ((HO)2(O)P—S-5′); any additional combination of oxygen/sulfur replaced monophosphate, diphosphate and triphosphates (e.g. 5′-alpha-thiotriphosphate, 5′-gamma-thiotriphosphate, etc.), 5′-phosphoramidates ((HO)2(O)P—NH-5′, (HO)(NH2)(O)P—O-5′), 5′-alkylphosphonates (R=alkyl=methyl, ethyl, isopropyl, propyl, etc., e.g. RP(OH)(O)—O-5′-, (OH)2(O)P-5′-CH2-), 5′-alkyletherphosphonates (R=alkylether=methoxymethyl (MeOCH2-), ethoxymethyl, etc., e.g. RP(OH)(O)—O-5′-). 
     The sense strand can be modified in order to inactivate the sense strand and prevent formation of an active RISC, thereby potentially reducing off-target effects. This can be accomplished by a modification which prevents 5′-phosphorylation of the sense strand, e.g., by modification with a 5′-O-methyl ribonucleotide (see Nykänen et al., (2001) ATP requirements and small interfering RNA structure in the RNA interference pathway. Cell 107, 309-321.) Other modifications which prevent phosphorylation can also be used, e.g., simply substituting the 5′-OH by H rather than 0-Me. Alternatively, a large bulky group may be added to the 5′-phosphate turning it into a phosphodiester linkage. 
     Non-Natural Nucleobases 
     Nitropyrrolyl and nitroindolyl are non-natural nucleobases that are members of a class of compounds known as universal bases. Universal bases are those compounds that can replace any of the four naturally occurring bases without substantially affecting the melting behavior or activity of the oligonucleotide duplex. In contrast to the stabilizing, hydrogen-bonding interactions associated with naturally occurring nucleobases, it is postulated that oligonucleotide duplexes containing 3-nitropyrrolyl nucleobases are stabilized solely by stacking interactions. The absence of significant hydrogen-bonding interactions with nitropyrrolyl nucleobases obviates the specificity for a specific complementary base. In addition, various reports confirm that 4-, 5- and 6-nitroindolyl display very little specificity for the four natural bases. Interestingly, an oligonucleotide duplex containing 5-nitroindolyl was more stable than the corresponding oligonucleotides containing 4-nitroindolyl and 6-nitroindolyl. Procedures for the preparation of 1-(2′-O-methyl-β-D-ribofuranosyl)-5-nitroindole are described in Gaubert, G.; Wengel, J.  Tetrahedron Letters  2004, 45, 5629. Other universal bases amenable to the present invention include hypoxanthinyl, isoinosinyl, 2-aza-inosinyl, 7-deaza-inosinyl, nitroimidazolyl, nitropyrazolyl, nitrobenzimidazolyl, nitroindazolyl, aminoindolyl, pyrrolopyrimidinyl, and structural derivatives thereof. For a more detailed discussion, including synthetic procedures, of nitropyrrolyl, nitroindolyl, and other universal bases mentioned above see Vallone et al., Nucleic Acids Research, 27(17):3589-3596 (1999); Loakes et al., J. Mol. Bio., 270:426-436 (1997); Loakes et al., Nucleic Acids Research, 22(20):4039-4043 (1994); Oliver et al., Organic Letters, Vol. 3(13):1977-1980 (2001); Amosova et al., Nucleic Acids Research, 25(10):1930-1934 (1997); Loakes et al., Nucleic Acids Research, 29(12):2437-2447 (2001); Bergstrom et al., J. Am. Chem. Soc., 117:1201-1209 (1995); Franchetti et al., Biorg. Med. Chem. Lett. 11:67-69 (2001); and Nair et al., Nucelosides, Nucleotides &amp; Nucleic Acids, 20(4-7):735-738 (2001). 
     Difluorotolyl is a non-natural nucleobase that functions as a universal base. Difluorotolyl is an isostere of the natural nucleobase thymine. But unlike thymine, difluorotolyl shows no appreciable selectivity for any of the natural bases. Other aromatic compounds that function as universal bases and are amenable to the present invention are 4-fluoro-6-methylbenzimidazole and 4-methylbenzimidazole. In addition, the relatively hydrophobic isocarbostyrilyl derivatives 3-methyl isocarbostyrilyl, 5-methyl isocarbostyrilyl, and 3-methyl-7-propynyl isocarbostyrilyl are universal bases which cause only slight destabilization of oligonucleotide duplexes compared to the oligonucleotide sequence containing only natural bases. Other non-natural nucleobases contemplated in the present invention include 7-azaindolyl, 6-methyl-7-azaindolyl, imidizopyridinyl, 9-methyl-imidizopyridinyl, pyrrolopyrizinyl, isocarbostyrilyl, 7-propynyl isocarbostyrilyl, propynyl-7-azaindolyl, 2,4,5-trimethylphenyl, 4-methylindolyl, 4,6-dimethylindolyl, phenyl, napthalenyl, anthracenyl, phenanthracenyl, pyrenyl, stilbenyl, tetracenyl, pentacenyl, and structural derivates thereof. For a more detailed discussion, including synthetic procedures, of difluorotolyl, 4-fluoro-6-methylbenzimidazole, 4-methylbenzimidazole, and other non-natural bases mentioned above, see: Schweitzer et al., J. Org. Chem., 59:7238-7242 (1994); Berger et al., Nucleic Acids Research, 28(15):2911-2914 (2000); Moran et al., J. Am. Chem. Soc., 119:2056-2057 (1997); Morales et al., J. Am. Chem. Soc., 121:2323-2324 (1999); Guckian et al., J. Am. Chem. Soc., 118:8182-8183 (1996); Morales et al., J. Am. Chem. Soc., 122(6):1001-1007 (2000); McMinn et al., J. Am. Chem. Soc., 121:11585-11586 (1999); Guckian et al., J. Org. Chem., 63:9652-9656 (1998); Moran et al., Proc. Natl. Acad. Sci., 94:10506-10511 (1997); Das et al., J. Chem. Soc., Perkin Trans., 1:197-206 (2002); Shibata et al., J. Chem. Soc., Perkin Trans., 1:1605-1611 (2001); Wu et al., J. Am. Chem. Soc., 122(32):7621-7632 (2000); O&#39;Neill et al., J. Org. Chem., 67:5869-5875 (2002); Chaudhuri et al., J. Am. Chem. Soc., 117:10434-10442 (1995); and U.S. Pat. No. 6,218,108. 
     Transport of iRNA Agents into Cells 
     Not wishing to be bound by any theory, the chemical similarity between cholesterol-conjugated iRNA agents and certain constituents of lipoproteins (e.g. cholesterol, cholesteryl esters, phospholipids) may lead to the association of iRNA agents with lipoproteins (e.g. LDL, HDL) in blood and/or the interaction of the iRNA agent with cellular components having an affinity for cholesterol, e.g. components of the cholesterol transport pathway. Lipoproteins as well as their constituents are taken up and processed by cells by various active and passive transport mechanisms, for example, without limitation, endocytosis of LDL-receptor bound LDL, endocytosis of oxidized or otherwise modified LDLs through interaction with Scavenger receptor A, Scavenger receptor B1-mediated uptake of HDL cholesterol in the liver, pinocytosis, or transport of cholesterol across membranes by ABC (ATP-binding cassette) transporter proteins, e.g. ABC-A1, ABC-G1 or ABC-G4. Hence, cholesterol-conjugated iRNA agents could enjoy facilitated uptake by cells possessing such transport mechanisms, e.g. cells of the liver. As such, the present invention provides evidence and general methods for targeting iRNA agents to cells expressing certain cell surface components, e.g. receptors, by conjugating a natural ligand for such component (e.g. cholesterol) to the iRNA agent, or by conjugating a chemical moiety (e.g. cholesterol) to the iRNA agent which associates with or binds to a natural ligand for the component (e.g. LDL, HDL). 
     Other Embodiments 
     An iRNA agent, can be produced in a cell in vivo, e.g., from exogenous DNA templates that are delivered into the cell. For example, the DNA templates can be inserted into vectors and used as gene therapy vectors. Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (U.S. Pat. No. 5,328,470), or by stereotactic injection (see, e.g., Chen et al. Proc. Natl. Acad. Sci. USA 91:3054-3057, 1994). The pharmaceutical preparation of the gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. The DNA templates, for example, can include two transcription units, one that produces a transcript that includes the top strand of an iRNA agent and one that produces a transcript that includes the bottom strand of an iRNA agent. When the templates are transcribed, the iRNA agent is produced, and processed into siRNA agent fragments that mediate gene silencing. 
     Formulation 
     The present invention also includes pharmaceutical compositions and formulations which include the dsRNA compounds of the invention. The pharmaceutical compositions of the present invention may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical, pulmonary, e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal, oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration. 
     Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable. Coated condoms, gloves and the like may also be useful. Preferred topical formulations include those in which the dsRNAs of the invention are in admixture with a topical delivery agent such as lipids, liposomes, fatty acids, fatty acid esters, steroids, chelating agents and surfactants. Preferred lipids and liposomes include neutral (e.g. dioleoylphosphatidyl ethanolamine=DOPE, dimyristoylphosphatidyl choline=DMPC, distearolyphosphatidyl choline) negative (e.g. dimyristoylphosphatidyl glycerol=DMPG) and cationic (e.g. dioleoyltetramethylaminopropyl=DOTAP and dioleoylphosphatidyl ethanolamine=DOTMA), e.g. (+/−)-N-(3-aminopropyl)-N,N-dimethyl-2,3-bis (dodecyloxy)-1-propanaminium bromide=GAP-DLRIE). DsRNAs of the invention may be encapsulated within liposomes or may form complexes thereto, in particular to cationic liposomes. Alternatively, dsRNAs may be complexed to lipids, in particular to cationic lipids. Preferred fatty acids and esters include but are not limited arachidonic acid, oleic acid, eicosanoic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein, dilaurin, glyceryl 1-monocaprate, 1-dodecylazacycloheptan-2-one, an acylcarnitine, an acylcholine, or a C 1-10  alkyl ester (e.g. isopropylmyristate IPM), monoglyceride, diglyceride or pharmaceutically acceptable salt thereof. Topical formulations are described in detail in U.S. patent application Ser. No. 09/315,298 filed on May 20, 1999 which is incorporated herein by reference in its entirety. 
     Compositions and formulations for oral administration include powders or granules, microparticulates, nanoparticulates, suspensions or solutions in water or non-aqueous media, capsules, gel capsules, sachets, tablets or minitablets. Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders may be desirable. Preferred oral formulations are those in which dsRNAs of the invention are administered in conjunction with one or more penetration enhancers, surfactants, and chelators. Preferred surfactants include fatty acids and/or esters or salts thereof, bile acids and/or salts thereof. Preferred bile acids/salts include chenodeoxycholic acid (CDCA) and ursodeoxychenodeoxycholic acid (UDCA), cholic acid, dehydrocholic acid, deoxycholic acid, glucholic acid, glycholic acid, glycodeoxycholic acid, taurocholic acid, taurodeoxycholic acid, sodium tauro-24,25-dihydro-fusidate and sodium glycodihydrofusidate. Preferred fatty acids include arachidonic acid, undecanoic acid, oleic acid, lauric acid, caprylic acid, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein, dilaurin, glyceryl 1-monocaprate, 1-dodecylazacycloheptan-2-one, an acylcarnitine, an acylcholine, or a monoglyceride, a diglyceride or a pharmaceutically acceptable salt thereof (e.g. sodium). Also preferred are combinations of penetration enhancers, for example, fatty acids/salts in combination with bile acids/salts. A particularly preferred combination is the sodium salt of lauric acid, capric acid and UDCA. Further penetration enhancers include polyoxyethylene-9-lauryl ether, polyoxyethylene-20-cetyl ether. DsRNAs of the invention may be delivered orally, in granular form including sprayed dried particles, or complexed to form micro or nanoparticles. DsRNA complexing agents include poly-amino acids; polyimines; polyacrylates; polyalkylacrylates, polyoxethanes, polyalkylcyanoacrylates; cationized gelatins, albumins, starches, acrylates, polyethyleneglycols (PEG) and starches; polyalkylcyanoacrylates; DEAE-derivatized polyimines, pollulans, celluloses and starches. Particularly preferred complexing agents include chitosan, N-trimethylchitosan, poly-L-lysine, polyhistidine, polyornithine, polyspermines, protamine, polyvinylpyridine, polythiodiethylaminomethylethylene P(TDAE), polyaminostyrene (e.g. p-amino), poly(methylcyanoacrylate), poly(ethylcyanoacrylate), poly(butylcyanoacrylate), poly(isobutylcyanoacrylate), poly(isohexylcynaoacrylate), DEAE-methacrylate, DEAE-hexylacrylate, DEAE-acrylamide, DEAE-albumin and DEAE-dextran, polymethylacrylate, polyhexylacrylate, poly(D,L-lactic acid), poly(DL-lactic-co-glycolic acid (PLGA), alginate, and polyethyleneglycol (PEG). Oral formulations for dsRNAs and their preparation are described in detail in U.S. application. Ser. No. 08/886,829 (filed Jul. 1, 1997), Ser. No. 09/108,673 (filed Jul. 1, 1998), Ser. No. 09/256,515 (filed Feb. 23, 1999), Ser. No. 09/082,624 (filed May 21, 1998) and Ser. No. 09/315,298 (filed May 20, 1999), each of which is incorporated herein by reference in their entirety. 
     Compositions and formulations for parenteral, intrathecal or intraventricular administration may include sterile aqueous solutions which may also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients. 
     Pharmaceutical compositions of the present invention include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions may be generated from a variety of components that include, but are not limited to, preformed liquids, self-emulsifying solids and self-emulsifying semisolids. 
     The pharmaceutical formulations of the present invention, which may conveniently be presented in unit dosage form, may be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s). In general, the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product. 
     The compositions of the present invention may be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, gel capsules, liquid syrups, soft gels, suppositories, and enemas. The compositions of the present invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media. Aqueous suspensions may further contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and/or dextran. The suspension may also contain stabilizers. 
     In one embodiment of the present invention the pharmaceutical compositions may be formulated and used as foams. Pharmaceutical foams include formulations such as, but not limited to, emulsions, microemulsions, creams, jellies and liposomes. While basically similar in nature these formulations vary in the components and the consistency of the final product. The preparation of such compositions and formulations is generally known to those skilled in the pharmaceutical and formulation arts and may be applied to the formulation of the compositions of the present invention. 
     Emulsions 
     The compositions of the present invention may be prepared and formulated as emulsions. Emulsions are typically heterogenous systems of one liquid dispersed in another in the form of droplets usually exceeding 0.1 μm in diameter (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199; Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., Volume 1, p. 245; Block in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 2, p. 335; Higuchi et al., in Remington&#39;s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1985, p. 301). Emulsions are often biphasic systems comprising two immiscible liquid phases intimately mixed and dispersed with each other. In general, emulsions may be of either the water-in-oil (w/o) or the oil-in-water (o/w) variety. When an aqueous phase is finely divided into and dispersed as minute droplets into a bulk oily phase, the resulting composition is called a water-in-oil (w/o) emulsion. Alternatively, when an oily phase is finely divided into and dispersed as minute droplets into a bulk aqueous phase, the resulting composition is called an oil-in-water (o/w) emulsion. Emulsions may contain additional components in addition to the dispersed phases, and the active drug which may be present as a solution in either the aqueous phase, oily phase or itself as a separate phase. Pharmaceutical excipients such as emulsifiers, stabilizers, dyes, and anti-oxidants may also be present in emulsions as needed. Pharmaceutical emulsions may also be multiple emulsions that are comprised of more than two phases such as, for example, in the case of oil-in-water-in-oil (o/w/o) and water-in-oil-in-water (w/o/w) emulsions. Such complex formulations often provide certain advantages that simple binary emulsions do not. Multiple emulsions in which individual oil droplets of an o/w emulsion enclose small water droplets constitute a w/o/w emulsion. Likewise a system of oil droplets enclosed in globules of water stabilized in an oily continuous phase provides an o/w/o emulsion. 
     Emulsions are characterized by little or no thermodynamic stability. Often, the dispersed or discontinuous phase of the emulsion is well dispersed into the external or continuous phase and maintained in this form through the means of emulsifiers or the viscosity of the formulation. Either of the phases of the emulsion may be a semisolid or a solid, as is the case of emulsion-style ointment bases and creams. Other means of stabilizing emulsions entail the use of emulsifiers that may be incorporated into either phase of the emulsion. Emulsifiers may broadly be classified into four categories: synthetic surfactants, naturally occurring emulsifiers, absorption bases, and finely dispersed solids (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). 
     Synthetic surfactants, also known as surface active agents, have found wide applicability in the formulation of emulsions and have been reviewed in the literature (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), Marcel Dekker, Inc., New York, N.Y., 1988, volume 1, p. 199). Surfactants are typically amphiphilic and comprise a hydrophilic and a hydrophobic portion. The ratio of the hydrophilic to the hydrophobic nature of the surfactant has been termed the hydrophile/lipophile balance (HLB) and is a valuable tool in categorizing and selecting surfactants in the preparation of formulations. Surfactants may be classified into different classes based on the nature of the hydrophilic group: nonionic, anionic, cationic and amphoteric (Rieger, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 285). 
     Naturally occurring emulsifiers used in emulsion formulations include lanolin, beeswax, phosphatides, lecithin and acacia. Absorption bases possess hydrophilic properties such that they can soak up water to form w/o emulsions yet retain their semisolid consistencies, such as anhydrous lanolin and hydrophilic petrolatum. Finely divided solids have also been used as good emulsifiers especially in combination with surfactants and in viscous preparations. These include polar inorganic solids, such as heavy metal hydroxides, nonswelling clays such as bentonite, attapulgite, hectorite, kaolin, montmorillonite, colloidal aluminum silicate and colloidal magnesium aluminum silicate, pigments and nonpolar solids such as carbon or glyceryl tristearate. 
     A large variety of non-emulsifying materials are also included in emulsion formulations and contribute to the properties of emulsions. These include fats, oils, waxes, fatty acids, fatty alcohols, fatty esters, humectants, hydrophilic colloids, preservatives and antioxidants (Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 335; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). 
     Hydrophilic colloids or hydrocolloids include naturally occurring gums and synthetic polymers such as polysaccharides (for example, acacia, agar, alginic acid, carrageenan, guar gum, karaya gum, and tragacanth), cellulose derivatives (for example, carboxymethylcellulose and carboxypropylcellulose), and synthetic polymers (for example, carbomers, cellulose ethers, and carboxyvinyl polymers). These disperse or swell in water to form colloidal solutions that stabilize emulsions by forming strong interfacial films around the dispersed-phase droplets and by increasing the viscosity of the external phase. 
     Since emulsions often contain a number of ingredients such as carbohydrates, proteins, sterols and phosphatides that may readily support the growth of microbes, these formulations often incorporate preservatives. Commonly used preservatives included in emulsion formulations include methyl paraben, propyl paraben, quaternary ammonium salts, benzalkonium chloride, esters of p-hydroxybenzoic acid, and boric acid. Antioxidants are also commonly added to emulsion formulations to prevent deterioration of the formulation. Antioxidants used may be free radical scavengers such as tocopherols, alkyl gallates, butylated hydroxyanisole, butylated hydroxytoluene, or reducing agents such as ascorbic acid and sodium metabisulfite, and antioxidant synergists such as citric acid, tartaric acid, and lecithin. 
     The application of emulsion formulations via dermatological, oral and parenteral routes and methods for their manufacture have been reviewed in the literature (Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). Emulsion formulations for oral delivery have been very widely used because of ease of formulation, as well as efficacy from an absorption and bioavailability standpoint (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245; Idson, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 199). Mineral-oil base laxatives, oil-soluble vitamins and high fat nutritive preparations are among the materials that have commonly been administered orally as o/w emulsions. 
     In one embodiment of the present invention, the compositions of dsRNAs and nucleic acids are formulated as microemulsions. A microemulsion may be defined as a system of water, oil and amphiphile which is a single optically isotropic and thermodynamically stable liquid solution (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245). Typically microemulsions are systems that are prepared by first dispersing an oil in an aqueous surfactant solution and then adding a sufficient amount of a fourth component, generally an intermediate chain-length alcohol to form a transparent system. Therefore, microemulsions have also been described as thermodynamically stable, isotropically clear dispersions of two immiscible liquids that are stabilized by interfacial films of surface-active molecules (Leung and Shah, in: Controlled Release of Drugs: Polymers and Aggregate Systems, Rosoff, M., Ed., 1989, VCH Publishers, New York, pages 185-215). Microemulsions commonly are prepared via a combination of three to five components that include oil, water, surfactant, cosurfactant and electrolyte. Whether the microemulsion is of the water-in-oil (w/o) or an oil-in-water (o/w) type is dependent on the properties of the oil and surfactant used and on the structure and geometric packing of the polar heads and hydrocarbon tails of the surfactant molecules (Schott, in Remington&#39;s Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 1985, p. 271). 
     The phenomenological approach utilizing phase diagrams has been extensively studied and has yielded a comprehensive knowledge, to one skilled in the art, of how to formulate microemulsions (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245; Block, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 335). Compared to conventional emulsions, microemulsions offer the advantage of solubilizing water-insoluble drugs in a formulation of thermodynamically stable droplets that are formed spontaneously. 
     Surfactants used in the preparation of microemulsions include, but are not limited to, ionic surfactants, non-ionic surfactants, Brij 96, polyoxyethylene oleyl ethers, polyglycerol fatty acid esters, tetraglycerol monolaurate (ML310), tetraglycerol monooleate (MO310), hexaglycerol monooleate (PO310), hexaglycerol pentaoleate (PO500), decaglycerol monocaprate (MCA750), decaglycerol monooleate (MO750), decaglycerol sequioleate (SO750), decaglycerol decaoleate (DAO750), alone or in combination with cosurfactants. The cosurfactant, usually a short-chain alcohol such as ethanol, 1-propanol, and 1-butanol, serves to increase the interfacial fluidity by penetrating into the surfactant film and consequently creating a disordered film because of the void space generated among surfactant molecules. Microemulsions may, however, be prepared without the use of cosurfactants and alcohol-free self-emulsifying microemulsion systems are known in the art. The aqueous phase may typically be, but is not limited to, water, an aqueous solution of the drug, glycerol, PEG300, PEG400, polyglycerols, propylene glycols, and derivatives of ethylene glycol. The oil phase may include, but is not limited to, materials such as Captex 300, Captex 355, Capmul MCM, fatty acid esters, medium chain (C 8 -C 12 ) mono, di, and tri-glycerides, polyoxyethylated glyceryl fatty acid esters, fatty alcohols, polyglycolized glycerides, saturated polyglycolized C 8 -C 10  glycerides, vegetable oils and silicone oil. 
     Microemulsions are particularly of interest from the standpoint of drug solubilization and the enhanced absorption of drugs. Lipid based microemulsions (both o/w and w/o) have been proposed to enhance the oral bioavailability of drugs, including peptides (Constantinides et al., Pharmaceutical Research, 1994, 11, 1385-1390; Ritschel, Meth. Find. Exp. Clin. Pharmacol., 1993, 13, 205). Microemulsions afford advantages of improved drug solubilization, protection of drug from enzymatic hydrolysis, possible enhancement of drug absorption due to surfactant-induced alterations in membrane fluidity and permeability, ease of preparation, ease of oral administration over solid dosage forms, improved clinical potency, and decreased toxicity (Constantinides et al., Pharmaceutical Research, 1994, 11, 1385; Ho et al., J. Pharm. Sci., 1996, 85, 138-143). Often microemulsions may form spontaneously when their components are brought together at ambient temperature. This may be particularly advantageous when formulating thermolabile drugs, peptides or dsRNAs. Microemulsions have also been effective in the transdermal delivery of active components in both cosmetic and pharmaceutical applications. It is expected that the microemulsion compositions and formulations of the present invention will facilitate the increased systemic absorption of dsRNAs and nucleic acids from the gastrointestinal tract, as well as improve the local cellular uptake of dsRNAs and nucleic acids within the gastrointestinal tract, vagina, buccal cavity and other areas of administration. 
     Microemulsions of the present invention may also contain additional components and additives such as sorbitan monostearate (Grill 3), Labrasol, and penetration enhancers to improve the properties of the formulation and to enhance the absorption of the dsRNAs and nucleic acids of the present invention. Penetration enhancers used in the microemulsions of the present invention may be classified as belonging to one of five broad categories-surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92). Each of these classes has been discussed above. 
     Liposomes 
     There are many organized surfactant structures besides microemulsions that have been studied and used for the formulation of drugs. These include monolayers, micelles, bilayers and vesicles. Vesicles, such as liposomes, have attracted great interest because of their specificity and the duration of action they offer from the standpoint of drug delivery. As used in the present invention, the term “liposome” means a vesicle composed of amphiphilic lipids arranged in a spherical bilayer or bilayers. 
     Liposomes are unilamellar or multilamellar vesicles which have a membrane formed from a lipophilic material and an aqueous interior. The aqueous portion contains the composition to be delivered. Cationic liposomes possess the advantage of being able to fuse to the cell wall. Non-cationic liposomes, although not able to fuse as efficiently with the cell membrane, are taken up by macrophages in vivo. 
     In order to cross intact mammalian skin, lipid vesicles must pass through a series of fine pores, each with a diameter less than 50 nm, under the influence of a suitable transdermal gradient. Therefore, it is desirable to use a liposome which is highly deformable and able to pass through such fine pores. 
     Further advantages of liposomes include; liposomes obtained from natural phospholipids are biocompatible and biodegradable; liposomes can incorporate a wide range of water and lipid soluble drugs; liposomes can protect encapsulated drugs in their internal compartments from metabolism and degradation (Rosoff, in Pharmaceutical Dosage Forms, Lieberman, Rieger and Banker (Eds.), 1988, Marcel Dekker, Inc., New York, N.Y., volume 1, p. 245). Important considerations in the preparation of liposome formulations are the lipid surface charge, vesicle size and the aqueous volume of the liposomes. 
     Liposomes are useful for the transfer and delivery of active ingredients to the site of action. Because the liposomal membrane is structurally similar to biological membranes, when liposomes are applied to a tissue, the liposomes start to merge with the cellular membranes and as the merging of the liposome and cell progresses, the liposomal contents are emptied into the cell where the active agent may act. 
     Liposomal formulations have been the focus of extensive investigation as the mode of delivery for many drugs. There is growing evidence that for topical administration, liposomes present several advantages over other formulations. Such advantages include reduced side-effects related to high systemic absorption of the administered drug, increased accumulation of the administered drug at the desired target, and the ability to administer a wide variety of drugs, both hydrophilic and hydrophobic, into the skin. 
     Several reports have detailed the ability of liposomes to deliver agents including high-molecular weight DNA into the skin. Compounds including analgesics, antibodies, hormones and high-molecular weight DNAs have been administered to the skin. The majority of applications resulted in the targeting of the upper epidermis 
     Liposomes fall into two broad classes. Cationic liposomes are positively charged liposomes which interact with the negatively charged DNA molecules to form a stable complex. The positively charged DNA/liposome complex binds to the negatively charged cell surface and is internalized in an endosome. Due to the acidic pH within the endosome, the liposomes are ruptured, releasing their contents into the cell cytoplasm (Wang et al., Biochem. Biophys. Res. Commun., 1987, 147, 980-985). 
     Liposomes which are pH-sensitive or negatively-charged, entrap DNA rather than complex with it. Since both the DNA and the lipid are similarly charged, repulsion rather than complex formation occurs. Nevertheless, some DNA is entrapped within the aqueous interior of these liposomes. pH-sensitive liposomes have been used to deliver DNA encoding the thymidine kinase gene to cell monolayers in culture. Expression of the exogenous gene was detected in the target cells (Zhou et al., Journal of Controlled Release, 1992, 19, 269-274). 
     One major type of liposomal composition includes phospholipids other than naturally derived phosphatidylcholine. Neutral liposome compositions, for example, can be formed from dimyristoyl phosphatidylcholine (DMPC) or dipalmitoyl phosphatidylcholine (DPPC). Anionic liposome compositions generally are formed from dimyristoyl phosphatidylglycerol, while anionic fusogenic liposomes are formed primarily from dioleoyl phosphatidylethanolamine (DOPE). Another type of liposomal composition is formed from phosphatidylcholine (PC) such as, for example, soybean PC, and egg PC. Another type is formed from mixtures of phospholipid and/or phosphatidylcholine and/or cholesterol. 
     Several studies have assessed the topical delivery of liposomal drug formulations to the skin. Application of liposomes containing interferon to guinea pig skin resulted in a reduction of skin herpes sores while delivery of interferon via other means (e.g. as a solution or as an emulsion) were ineffective (Weiner et al., Journal of Drug Targeting, 1992, 2, 405-410). Further, an additional study tested the efficacy of interferon administered as part of a liposomal formulation to the administration of interferon using an aqueous system, and concluded that the liposomal formulation was superior to aqueous administration (du Plessis et al., Antiviral Research, 1992, 18, 259-265). 
     Non-ionic liposomal systems have also been examined to determine their utility in the delivery of drugs to the skin, in particular systems comprising non-ionic surfactant and cholesterol. Non-ionic liposomal formulations comprising Novasome™ I (glyceryl dilaurate/cholesterol/polyoxyethylene-10-stearyl ether) and Novasome™ II (glyceryl distearate/cholesterol/polyoxyethylene-10-stearyl ether) were used to deliver cyclosporin-A into the dermis of mouse skin. Results indicated that such non-ionic liposomal systems were effective in facilitating the deposition of cyclosporin-A into different layers of the skin (Hu et al. S.T.P.Pharma. Sci., 1994, 4, 6, 466). 
     Liposomes also include “sterically stabilized” liposomes, a term which, as used herein, refers to liposomes comprising one or more specialized lipids that, when incorporated into liposomes, result in enhanced circulation lifetimes relative to liposomes lacking such specialized lipids. Examples of sterically stabilized liposomes are those in which part of the vesicle-forming lipid portion of the liposome (A) comprises one or more glycolipids, such as monosialoganglioside G m 1, or (B) is derivatized with one or more hydrophilic polymers, such as a polyethylene glycol (PEG) moiety. While not wishing to be bound by any particular theory, it is thought in the art that, at least for sterically stabilized liposomes containing gangliosides, sphingomyelin, or PEG-derivatized lipids, the enhanced circulation half-life of these sterically stabilized liposomes derives from a reduced uptake into cells of the reticuloendothelial system (RES) (Allen et al., FEBS Letters, 1987, 223, 42; Wu et al., Cancer Research, 1993, 53, 3765). 
     Various liposomes comprising one or more glycolipids are known in the art. Papahadjopoulos et al. (Ann. N.Y. Acad. Sci., 1987, 507, 64) reported the ability of monosialoganglioside G m 1, galactocerebroside sulfate and phosphatidylinositol to improve blood half-lives of liposomes. These findings were expounded upon by Gabizon et al. (Proc. Natl. Acad. Sci. U.S.A., 1988, 85, 6949). U.S. Pat. No. 4,837,028 and WO 88/04924, both to Allen et al., disclose liposomes comprising (1) sphingomyelin and (2) the ganglioside G m 1 or a galactocerebroside sulfate ester. U.S. Pat. No. 5,543,152 (Webb et al.) discloses liposomes comprising sphingomyelin. Liposomes comprising 1,2-sn-dimyristoylphosphat-idylcholine are disclosed in WO 97/13499 (Lim et al). 
     Many liposomes comprising lipids derivatized with one or more hydrophilic polymers, and methods of preparation thereof, are known in the art. Sunamoto et al. (Bull. Chem. Soc. Jpn., 1980, 53, 2778) described liposomes comprising a nonionic detergent, 2C 1215G , that contains a PEG moiety. Illum et al. (FEBS Lett., 1984, 167, 79) noted that hydrophilic coating of polystyrene particles with polymeric glycols results in significantly enhanced blood half-lives. Synthetic phospholipids modified by the attachment of carboxylic groups of polyalkylene glycols (e.g., PEG) are described by Sears (U.S. Pat. Nos. 4,426,330 and 4,534,899). Klibanov et al. (FEBS Lett., 1990, 268, 235) described experiments demonstrating that liposomes comprising phosphatidylethanolamine (PE) derivatized with PEG or PEG stearate have significant increases in blood circulation half-lives. Blume et al. (Biochimica et Biophysica Acta, 1990, 1029, 91) extended such observations to other PEG-derivatized phospholipids, e.g., DSPE-PEG, formed from the combination of distearoylphosphatidylethanolamine (DSPE) and PEG. Liposomes having covalently bound PEG moieties on their external surface are described in European Patent No. EP 0 445 131 B1 and WO 90/04384 to Fisher. Liposome compositions containing 1-20 mole percent of PE derivatized with PEG, and methods of use thereof, are described by Woodle et al. (U.S. Pat. Nos. 5,013,556 and 5,356,633) and Martin et al. (U.S. Pat. No. 5,213,804 and European Patent No. EP 0 496 813 B1). Liposomes comprising a number of other lipid-polymer conjugates are disclosed in WO 91/05545 and U.S. Pat. No. 5,225,212 (both to Martin et al.) and in WO 94/20073 (Zalipsky et al.) Liposomes comprising PEG-modified ceramide lipids are described in WO 96/10391 (Choi et al). U.S. Pat. No. 5,540,935 (Miyazaki et al.) and U.S. Pat. No. 5,556,948 (Tagawa et al.) describe PEG-containing liposomes that can be further derivatized with functional moieties on their surfaces. 
     A limited number of liposomes comprising nucleic acids are known in the art. WO 96/40062 to Thierry et al. discloses methods for encapsulating high molecular weight nucleic acids in liposomes. U.S. Pat. No. 5,264,221 to Tagawa et al. discloses protein-bonded liposomes and asserts that the contents of such liposomes may include dsRNA. U.S. Pat. No. 5,665,710 to Rahman et al. describes certain methods of encapsulating oligodeoxynucleotides in liposomes. WO 97/04787 to Love et al. discloses liposomes comprising dsRNAs targeted to the raf gene. 
     Transfersomes are yet another type of liposomes, and are highly deformable lipid aggregates which are attractive candidates for drug delivery vehicles. Transfersomes may be described as lipid droplets which are so highly deformable that they are easily able to penetrate through pores which are smaller than the droplet. Transfersomes are adaptable to the environment in which they are used, e.g. they are self-optimizing (adaptive to the shape of pores in the skin), self-repairing, frequently reach their targets without fragmenting, and often self-loading. To make transfersomes it is possible to add surface edge-activators, usually surfactants, to a standard liposomal composition. Transfersomes have been used to deliver serum albumin to the skin. The transfersome-mediated delivery of serum albumin has been shown to be as effective as subcutaneous injection of a solution containing serum albumin. 
     Surfactants find wide application in formulations such as emulsions (including microemulsions) and liposomes. The most common way of classifying and ranking the properties of the many different types of surfactants, both natural and synthetic, is by the use of the hydrophile/lipophile balance (HLB). The nature of the hydrophilic group (also known as the “head”) provides the most useful means for categorizing the different surfactants used in formulations (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p. 285). 
     If the surfactant molecule is not ionized, it is classified as a nonionic surfactant. Nonionic surfactants find wide application in pharmaceutical and cosmetic products and are usable over a wide range of pH values. In general their HLB values range from 2 to about 18 depending on their structure. Nonionic surfactants include nonionic esters such as ethylene glycol esters, propylene glycol esters, glyceryl esters, polyglyceryl esters, sorbitan esters, sucrose esters, and ethoxylated esters. Nonionic alkanolamides and ethers such as fatty alcohol ethoxylates, propoxylated alcohols, and ethoxylated/propoxylated block polymers are also included in this class. The polyoxyethylene surfactants are the most popular members of the nonionic surfactant class. 
     If the surfactant molecule carries a negative charge when it is dissolved or dispersed in water, the surfactant is classified as anionic. Anionic surfactants include carboxylates such as soaps, acyl lactylates, acyl amides of amino acids, esters of sulfuric acid such as alkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkyl benzene sulfonates, acyl isethionates, acyl taurates and sulfosuccinates, and phosphates. The most important members of the anionic surfactant class are the alkyl sulfates and the soaps. 
     If the surfactant molecule carries a positive charge when it is dissolved or dispersed in water, the surfactant is classified as cationic. Cationic surfactants include quaternary ammonium salts and ethoxylated amines. The quaternary ammonium salts are the most used members of this class. 
     If the surfactant molecule has the ability to carry either a positive or negative charge, the surfactant is classified as amphoteric. Amphoteric surfactants include acrylic acid derivatives, substituted alkylamides, N-alkylbetaines and phosphatides. 
     The use of surfactants in drug products, formulations and in emulsions has been reviewed (Rieger, in Pharmaceutical Dosage Forms, Marcel Dekker, Inc., New York, N.Y., 1988, p. 285). 
     Penetration Enhancers 
     In one embodiment, the present invention employs various penetration enhancers to effect the efficient delivery of nucleic acids, particularly dsRNAs, to the skin of animals. Most drugs are present in solution in both ionized and nonionized forms. However, usually only lipid soluble or lipophilic drugs readily cross cell membranes. It has been discovered that even non-lipophilic drugs may cross cell membranes if the membrane to be crossed is treated with a penetration enhancer. In addition to aiding the diffusion of non-lipophilic drugs across cell membranes, penetration enhancers also enhance the permeability of lipophilic drugs. 
     Penetration enhancers may be classified as belonging to one of five broad categories, i.e., surfactants, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92). Each of the above mentioned classes of penetration enhancers are described below in greater detail. 
     Surfactants: In connection with the present invention, surfactants (or “surface-active agents”) are chemical entities which, when dissolved in an aqueous solution, reduce the surface tension of the solution or the interfacial tension between the aqueous solution and another liquid, with the result that absorption of dsRNAs through the mucosa is enhanced. In addition to bile salts and fatty acids, these penetration enhancers include, for example, sodium lauryl sulfate, polyoxyethylene-9-lauryl ether and polyoxyethylene-20-cetyl ether) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, p. 92); and perfluorochemical emulsions, such as FC-43 (Takahashi et al., J. Pharm. Pharmacol., 1988, 40, 252). 
     Fatty acids: Various fatty acids and their derivatives which act as penetration enhancers include, for example, oleic acid, lauric acid, capric acid (n-decanoic acid), myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, tricaprate, monoolein (1-monooleoyl-rac-glycerol), dilaurin, caprylic acid, arachidonic acid, glycerol 1-monocaprate, 1-dodecylazacycloheptan-2-one, acylcarnitines, acylcholines, C 1 -C 10  alkyl esters thereof (e.g., methyl, isopropyl and t-butyl), and mono- and di-glycerides thereof (i.e., oleate, laurate, caprate, myristate, palmitate, stearate, linoleate, etc.) (Lee et al., Critical Reviews in Therapeutic Drug Carryier Systems, 1991, p. 92; Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33; El Hariri et al., J. Pharm. Pharmacol., 1992, 44, 651-654). 
     Bile salts: The physiological role of bile includes the facilitation of dispersion and absorption of lipids and fat-soluble vitamins (Brunton, Chapter 38 in: Goodman &amp; Gilman&#39;s The Pharmacological Basis of Therapeutics, 9th Ed., Hardman et al. Eds., McGraw-Hill, New York, 1996, pp. 934-935). Various natural bile salts, and their synthetic derivatives, act as penetration enhancers. Thus the term “bile salts” includes any of the naturally occurring components of bile as well as any of their synthetic derivatives. The bile salts of the invention include, for example, cholic acid (or its pharmaceutically acceptable sodium salt, sodium cholate), dehydrocholic acid (sodium dehydrocholate), deoxycholic acid (sodium deoxycholate), glucholic acid (sodium glucholate), glycholic acid (sodium glycocholate), glycodeoxycholic acid (sodium glycodeoxycholate), taurocholic acid (sodium taurocholate), taurodeoxycholic acid (sodium taurodeoxycholate), chenodeoxycholic acid (sodium chenodeoxycholate), ursodeoxycholic acid (UDCA), sodium tauro-24,25-dihydro-fusidate (STDHF), sodium glycodihydrofusidate and polyoxyethylene-9-lauryl ether (POE) (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92; Swinyard, Chapter 39 In: Remington&#39;s Pharmaceutical Sciences, 18th Ed., Gennaro, ed., Mack Publishing Co., Easton, Pa., 1990, pages 782-783; Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33; Yamamoto et al., J. Pharm. Exp. Ther., 1992, 263, 25; Yamashita et al., J. Pharm. Sci., 1990, 79, 579-583). 
     Chelating Agents: Chelating agents, as used in connection with the present invention, can be defined as compounds that remove metallic ions from solution by forming complexes therewith, with the result that absorption of dsRNAs through the mucosa is enhanced. With regards to their use as penetration enhancers in the present invention, chelating agents have the added advantage of also serving as DNase inhibitors, as most characterized DNA nucleases require a divalent metal ion for catalysis and are thus inhibited by chelating agents (Jarrett, J. Chromatogr., 1993, 618, 315-339). Chelating agents of the invention include but are not limited to disodium ethylenediaminetetraacetate (EDTA), citric acid, salicylates (e.g., sodium salicylate, 5-methoxysalicylate and homovanilate), N-acyl derivatives of collagen, laureth-9 and N-amino acyl derivatives of beta-diketones (enamines)(Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92; Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33; Buur et al., J. Control Rel., 1990, 14, 43-51). 
     Non-chelating non-surfactants: As used herein, non-chelating non-surfactant penetration enhancing compounds can be defined as compounds that demonstrate insignificant activity as chelating agents or as surfactants but that nonetheless enhance absorption of dsRNAs through the alimentary mucosa (Muranishi, Critical Reviews in Therapeutic Drug Carrier Systems, 1990, 7, 1-33). This class of penetration enhancers include, for example, unsaturated cyclic ureas, 1-alkyl- and 1-alkenylazacyclo-alkanone derivatives (Lee et al., Critical Reviews in Therapeutic Drug Carrier Systems, 1991, page 92); and non-steroidal anti-inflammatory agents such as diclofenac sodium, indomethacin and phenylbutazone (Yamashita et al., J. Pharm. Pharmacol., 1987, 39, 621-626). 
     Agents that enhance uptake of dsRNAs at the cellular level may also be added to the pharmaceutical and other compositions of the present invention. For example, cationic lipids, such as lipofectin (Junichi et al, U.S. Pat. No. 5,705,188), cationic glycerol derivatives, and polycationic molecules, such as polylysine (Lollo et al., PCT Application WO 97/30731) and other peptides, are also known to enhance the cellular uptake of dsRNAs. 
     Other agents may be utilized to enhance the penetration of the administered nucleic acids, including glycols such as ethylene glycol and propylene glycol, pyrrols such as 2-pyrrol, azones, and terpenes such as limonene and menthone. 
     Carriers 
     Certain compositions of the present invention also incorporate carrier compounds in the formulation. As used herein, “carrier compound” or “carrier” can refer to a nucleic acid, or analog thereof, which is inert (i.e., does not possess biological activity per se) but is recognized as a nucleic acid by in vivo processes that reduce the bioavailability of a nucleic acid having biological activity by, for example, degrading the biologically active nucleic acid or promoting its removal from circulation. The coadministration of a nucleic acid and a carrier compound, typically with an excess of the latter substance, can result in a substantial reduction of the amount of nucleic acid recovered in the liver, kidney or other extracirculatory reservoirs, presumably due to competition between the carrier compound and the nucleic acid for a common receptor. For example, the recovery of a partially phosphorothioate dsRNA in hepatic tissue can be reduced when it is coadministered with polyinosinic acid, dextran sulfate, polycytidic acid or 4-acetamido-4′isothiocyano-stilbene-2,2′-disulfonic acid (Miyao et al., Antisense Res. Dev., 1995, 5, 115-121; Takakura et al., Antisense &amp; Nucl. Acid Drug Dev., 1996, 6, 177-183. 
     Excipients 
     In contrast to a carrier compound, a “pharmaceutical carrier” or “excipient” is a pharmaceutically acceptable solvent, suspending agent or any other pharmacologically inert vehicle for delivering one or more nucleic acids to an animal. The excipient may be liquid or solid and is selected, with the planned manner of administration in mind, so as to provide for the desired bulk, consistency, etc., when combined with a nucleic acid and the other components of a given pharmaceutical composition. Typical pharmaceutical carriers include, but are not limited to, binding agents (e.g., pregelatinized maize starch, polyvinylpyrrolidone or hydroxypropyl methylcellulose, etc.); fillers (e.g., lactose and other sugars, microcrystalline cellulose, pectin, gelatin, calcium sulfate, ethyl cellulose, polyacrylates or calcium hydrogen phosphate, etc.); lubricants (e.g., magnesium stearate, talc, silica, colloidal silicon dioxide, stearic acid, metallic stearates, hydrogenated vegetable oils, corn starch, polyethylene glycols, sodium benzoate, sodium acetate, etc.); disintegrants (e.g., starch, sodium starch glycolate, etc.); and wetting agents (e.g., sodium lauryl sulphate, etc). 
     Pharmaceutically acceptable organic or inorganic excipient suitable for non-parenteral administration which do not deleteriously react with nucleic acids can also be used to formulate the compositions of the present invention. Suitable pharmaceutically acceptable carriers include, but are not limited to, water, salt solutions, alcohols, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, polyvinylpyrrolidone and the like. 
     Formulations for topical administration of nucleic acids may include sterile and non-sterile aqueous solutions, non-aqueous solutions in common solvents such as alcohols, or solutions of the nucleic acids in liquid or solid oil bases. The solutions may also contain buffers, diluents and other suitable additives. Pharmaceutically acceptable organic or inorganic excipients suitable for non-parenteral administration which do not deleteriously react with nucleic acids can be used. 
     Suitable pharmaceutically acceptable excipients include, but are not limited to, water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylose, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulose, polyvinylpyrrolidone and the like. 
     Pharmaceutical Compositions for the Delivery to the Respiratory Tract 
     Another aspect of the invention provides for the delivery of iRNA agents to the respiratory tract, particularly for the treatment of cystic fibrosis. The respiratory tract includes the upper airways, including the oropharynx and larynx, followed by the lower airways, which include the trachea followed by bifurcations into the bronchi and bronchioli. The upper and lower airways are called the conductive airways. The terminal bronchioli then divide into respiratory bronchioli which then lead to the ultimate respiratory zone, the alveoli, or deep lung. The epithelium of the conductive airways is the primary target of inhaled therapeutic aerosols for delivery of iRNA agents such as alpha-ENaC iRNA agents. 
     Pulmonary delivery compositions can be delivered by inhalation by the patient of a dispersion so that the composition, preferably the iRNA agent, within the dispersion can reach the lung where it can, for example, be readily absorbed through the alveolar region directly into blood circulation. Pulmonary delivery can be effective both for systemic delivery and for localized delivery to treat diseases of the lungs. 
     Pulmonary delivery can be achieved by different approaches, including the use of nebulized, aerosolized, micellular and dry powder-based formulations; administration by inhalation may be oral and/or nasal. Delivery can be achieved with liquid nebulizers, aerosol-based inhalers, and dry powder dispersion devices. Metered-dose devices are preferred. One of the benefits of using an atomizer or inhaler is that the potential for contamination is minimized because the devices are self contained. Dry powder dispersion devices, for example, deliver drugs that may be readily formulated as dry powders. An iRNA composition may be stably stored as lyophilized or spray-dried powders by itself or in combination with suitable powder carriers. The delivery of a composition for inhalation can be mediated by a dosing timing element which can include a timer, a dose counter, time measuring device, or a time indicator which when incorporated into the device enables dose tracking, compliance monitoring, and/or dose triggering to a patient during administration of the aerosol medicament. 
     Examples of pharmaceutical devices for aerosol delivery include metered dose inhalers (MDIs), dry powder inhalers (DPIs), and air-jet nebulizers. Exemplary delivery systems by inhalation which can be readily adapted for delivery of the subject iRNA agents are described in, for example, U.S. Pat. Nos. 5,756,353; 5,858,784; and PCT applications WO98/31346; WO98/10796; WO00/27359; WO01/54664; WO02/060412. Other aerosol formulations that may be used for delivering the iRNA agents are described in U.S. Pat. Nos. 6,294,153; 6,344,194; 6,071,497, and PCT applications WO02/066078; WO02/053190; WO01/60420; WO00/66206. Further, methods for delivering iRNA agents can be adapted from those used in delivering other oligonucleotides (e.g., an antisense oligonucleotide) by inhalation, such as described in Templin et al., Antisense Nucleic Acid Drug Dev, 2000, 10:359-68; Sandrasagra et al., Expert Opin Biol Ther, 2001, 1:979-83; Sandrasagra et al., Antisense Nucleic Acid Drug Dev, 2002, 12:177-81. 
     The delivery of the inventive agents may also involve the administration of so called “pro-drugs”, i.e. formulations or chemical modifications of a therapeutic substance that require some form of processing or transport by systems innate to the subject organism to release the therapeutic substance, preferably at the site where its action is desired; this latter embodiment may be used in conjunction with delivery of the respiratory tract, but also together with other embodiments of the present invention. For example, the human lungs can remove or rapidly degrade hydrolytically cleavable deposited aerosols over periods ranging from minutes to hours. In the upper airways, ciliated epithelia contribute to the “mucociliary excalator” by which particles are swept from the airways toward the mouth. Pavia, D., “Lung Mucociliary Clearance,” in Aerosols and the Lung: Clinical and Experimental Aspects, Clarke, S. W. and Pavia, D., Eds., Butterworths, London, 1984. In the deep lungs, alveolar macrophages are capable of phagocytosing particles soon after their deposition. Warheit et al. Microscopy Res. Tech., 26: 412-422 (1993); and Brain, J. D., “Physiology and Pathophysiology of Pulmonary Macrophages,” in The Reticuloendothelial System, S. M. Reichard and J. Filkins, Eds., Plenum, New. York., pp. 315-327, 1985. 
     In preferred embodiments, particularly where systemic dosing with the iRNA agent is desired, the aerosoled iRNA agents are formulated as microparticles. Microparticles having a diameter of between 0.5 and ten microns can penetrate the lungs, passing through most of the natural barriers. A diameter of less than ten microns is required to bypass the throat; a diameter of 0.5 microns or greater is required to avoid being exhaled. 
     Other Components 
     Compositions of the present invention may additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art-established usage levels. Thus, for example, the compositions may contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti-inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the components of the compositions of the present invention. The formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings and/or aromatic substances and the like which do not deleteriously interact with the nucleic acid(s) of the formulation. 
     Aqueous suspensions may contain substances which increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and/or dextran. The suspension may also contain stabilizers. 
     Certain embodiments of the invention provide pharmaceutical combinations and compositions containing (a) one or more dsRNA agents and (b) one or more other therapeutic agents which function by a non-RNA interference mechanism. 
     Accordingly, the invention includes a combination of an iRNA of the present invention with an anti-inflammatory, bronchodilatory, antihistamine, anti-tussive, antibiotic or DNase drug substance, said epithelial sodium channel blocker and said drug substance being in the same or different pharmaceutical composition. 
     Suitable antibiotics include macrolide antibiotics, e.g., tobramycin (TOBI™). 
     Suitable DNase drug substances include dornase alfa (Pulmozyme™), a highly-purified solution of recombinant human deoxyribonuclease I (rhDNase), which selectively cleaves DNA. Dornase alfa is used to treat cystic fibrosis. 
     Other useful combinations of epithelial sodium channel blockers with anti-inflammatory drugs are those with antagonists of chemokine receptors, e.g., CCR-1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5 antagonists, such as Schering-Plough antagonists SC-351125, SCH-55700 and SCH-D; Takeda antagonists, such as N-[[4-[[[6,7-dihydro-2-(4-methyl-phenyl)-5H-benzo-cyclohepten-8-yl]carbonyl]amino]phenyl]-methyl]tetrahydro-N,N-dimethyl-2H-pyran-4-amin-ium chloride (TAK-770); and CCR-5 antagonists described in U.S. Pat. No. 6,166,037 (particularly claims 18 and 19), WO 00/66558 (particularly claim  8 ), WO 00/66559 (particularly claim  9 ), WO 04/018425 and WO 04/026873. 
     Suitable anti-inflammatory drugs include steroids, in particular, glucocorticosteroids, such as budesonide, beclamethasone dipropionate, fluticasone propionate, ciclesonide or mometasone furoate, or steroids described in WO 02/88167, WO 02/12266, WO 02/100879, WO 02/00679 (especially those of Examples 3, 11, 14, 17, 19, 26, 34, 37, 39, 51, 60, 67, 72, 73, 90, 99 and 101), WO 03/35668, WO 03/48181, WO 03/62259, WO 03/64445, WO 03/72592, WO 04/39827 and WO 04/66920; non-steroidal glucocorticoid receptor agonists, such as those described in DE 10261874, WO 00/00531, WO 02/10143, WO 03/82280, WO 03/82787, WO 03/86294, WO 03/104195, WO 03/101932, WO 04/05229, WO 04/18429, WO 04/19935 and WO 04/26248; LTD4 antagonists, such as montelukast and zafirlukast; PDE4 inhibitors, such as cilomilast (Ariflo® GlaxoSmithKline), Roflumilast (Byk Gulden), V-11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering-Plough), Arofylline (Almirall Prodesfarma), PD189659/PD168787 (Parke-Davis), AWD-12-281 (Asta Medica), CDC-801 (Celgene), SelCID™ CC-10004 (Celgene), VM554/UM565 (Vernalis), T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo), and those disclosed in WO 92/19594, WO 93/19749, WO 93/19750, WO 93/19751, WO 98/18796, WO 99/16766, WO 01/13953, WO 03/104204, WO 03/104205, WO 03/39544, WO 04/000814, WO 04/000839, WO 04/005258, WO 04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/018431, WO 04/018449, WO 04/018450, WO 04/018451, WO 04/018457, WO 04/018465, WO 04/019944, WO 04/019945, WO 04/045607 and WO 04/037805; adenosine A2B receptor antagonists such as those described in WO 02/42298; and beta-2 adrenoceptor agonists, such as albuterol (salbutamol), metaproterenol, terbutaline, salmeterol fenoterol, procaterol, and especially, formoterol, carmoterol and pharmaceutically acceptable salts thereof, and compounds (in free or salt or solvate form) of formula (I) of WO 0075114, which document is incorporated herein by reference, preferably compounds of the Examples thereof, especially indacaterol and pharmaceutically acceptable salts thereof, as well as compounds (in free or salt or solvate form) of formula (I) of WO 04/16601, and also compounds of EP 1440966, JP 05025045, WO 93/18007, WO 99/64035, USP 2002/0055651, WO 01/42193, WO 01/83462, WO 02/66422, WO 02/70490, WO 02/76933, WO 03/24439, WO 03/42160, WO 03/42164, WO 03/72539, WO 03/91204, WO 03/99764, WO 04/16578, WO 04/22547, WO 04/32921, WO 04/33412, WO 04/37768, WO 04/37773, WO 04/37807, WO 04/39762, WO 04/39766, WO 04/45618, WO 04/46083, WO 04/80964, WO 04/108765 and WO 04/108676. 
     Suitable bronchodilatory drugs include anticholinergic or antimuscarinic agents, in particular, ipratropium bromide, oxitropium bromide, tiotropium salts and CHF 4226 (Chiesi), and glycopyrrolate, but also those described in EP 424021, U.S. Pat. Nos. 3,714,357, 5,171,744, WO 01/04118, WO 02/00652, WO 02/51841, WO 02/53564, WO 03/00840, WO 03/33495, WO 03/53966, WO 03/87094, WO 04/018422 and WO 04/05285. 
     Suitable dual anti-inflammatory and bronchodilatory drugs include dual beta-2 adrenoceptor agonist/muscarinic antagonists such as those disclosed in USP 2004/0167167, WO 04/74246 and WO 04/74812. 
     Suitable antihistamine drug substances include cetirizine hydrochloride, acetaminophen, clemastine fumarate, promethazine, loratidine, desloratidine, diphenhydramine and fexofenadine hydrochloride, activastine, astemizole, azelastine, ebastine, epinastine, mizolastine and tefenadine, as well as those disclosed in JP 2004107299, WO 03/099807 and WO 04/026841. 
     Other useful combinations of agents of the invention with anti-inflammatory drugs are those with antagonists of chemokine receptors, e.g., CCR-1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5 antagonists, such as Schering-Plough antagonists SC-351125, SCH-55700 and SCH-D; Takeda antagonists, such as N-[[4-[[[6,7-dihydro-2-(4-methylphenyl)-5H-benzo-cyclohepten-8-yl]carbonyl]amino]phenyl]-methyl]tetrahydro-N,N-dimethyl-2H-pyran-4-amin-ium chloride (TAK-770), and CCR-5 antagonists described in U.S. Pat. No. 6,166,037 (particularly claims 18 and 19), WO 00/66558 (particularly claim  8 ), WO 00/66559 (particularly claim  9 ), WO 04/018425 and WO 04/026873. 
     Other useful additional therapeutic agents may also be selected from the group consisting of cytokine binding molecules, particularly antibodies of other cytokines, in particular a combination with an anti-IL4 antibody, such as described in PCT/EP2005/00836, an anti-IgE antibody, such as Xolair®, an anti-IL31 antibody, an anti-IL31R antibody, an anti-TSLP antibody, an anti-TSLP receptor antibody, an anti-endoglin antibody, an anti-IL1b antibody or an anti-IL13 antibody, such as described in WO05/007699. 
     Two or more combined compounds may be used together in a single formulation, separately, concomitantly or sequentially. 
     Toxicity and therapeutic efficacy of such compounds can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index and it can be expressed as the ratio LD50/ED50. Compounds which exhibit high therapeutic indices are preferred. 
     The data obtained from cell culture assays and animal studies can be used in formulating a range of dosage for use in humans. The dosage of compositions of the invention lies generally within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any compound used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range of the compound or, when appropriate, of the polypeptide product of a target sequence (e.g., achieving a decreased concentration of the polypeptide) that includes the IC50 (i.e., the concentration of the test compound which achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography. 
     In addition to their administration individually or as a plurality, as discussed above, the dsRNAs of the invention can be administered in combination with other known agents effective in treatment of ENaC related disorders. In any event, the administering physician can adjust the amount and timing of dsRNA administration on the basis of results observed using standard measures of efficacy known in the art or described herein. 
     Treatment Methods and Routes of Delivery 
     A composition that includes an iRNA agent, e.g., an iRNA agent that targets alpha-ENaC, can be delivered to a subject by a variety of routes to achieve either local delivery to the site of action or systemic delivery to the subject. Exemplary routes include direct local administration to the site of treatment, such as the lungs and nasal passage as well as intravenous, nasal, oral, and ocular delivery. The preferred means of administering the iRNA agents of the present invention is through direct administration to the lungs and nasal passage as a liquid, aerosol or nebulized solution. 
     Formulations for inhalation or parenteral administration are well known in the art. Such formulation may include sterile aqueous solutions which may also contain buffers, diluents and other suitable additives. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic. 
     The active compounds disclosed herein are preferably administered to the lung(s) or nasal passage of a subject by any suitable means. Active compounds may be administered by administering an aerosol suspension of respirable particles comprised of the active compound or active compounds, which the subject inhales. The active compound can be aerosolized in a variety of forms, such as, but not limited to, dry powder inhalants, metered dose inhalants, or liquid/liquid suspensions. The respirable particles may be liquid or solid. The particles may optionally contain other therapeutic ingredients such as amiloride, benzamil or phenamil, with the selected compound included in an amount effective to inhibit the reabsorption of water from airway mucous secretions, as described in U.S. Pat. No. 4,501,729. 
     The particulate pharmaceutical composition may optionally be combined with a carrier to aid in dispersion or transport. A suitable carrier such as a sugar (i.e., lactose, sucrose, trehalose, mannitol) may be blended with the active compound or compounds in any suitable ratio (e.g., a 1 to 1 ratio by weight). 
     Particles comprised of the active compound for practicing the present invention should include particles of respirable size, that is, particles of a size sufficiently small to pass through the mouth or nose and larynx upon inhalation and into the bronchi and alveoli of the lungs. In general, particles ranging from about 1 to 10 microns in size (more particularly, less than about 5 microns in size) are respirable. Particles of non-respirable size which are included in the aerosol tend to deposit in the throat and be swallowed, and the quantity of non-respirable particles in the aerosol is preferably minimized. For nasal administration, a particle size in the range of 10-500 uM is preferred to ensure retention in the nasal cavity. 
     Liquid pharmaceutical compositions of active compound for producing an aerosol may be prepared by combining the active compound with a suitable vehicle, such as sterile pyrogen free water. The hypertonic saline solutions used to carry out the present invention are preferably sterile, pyrogen-free solutions, comprising from one to fifteen percent (by weight) of the physiologically acceptable salt, and more preferably from three to seven percent by weight of the physiologically acceptable salt. 
     Aerosols of liquid particles comprising the active compound may be produced by any suitable means, such as with a pressure-driven jet nebulizer or an ultrasonic nebulizer. See, e.g., U.S. Pat. No. 4,501,729. Nebulizers are commercially available devices which transform solutions or suspensions of the active ingredient into a therapeutic aerosol mist either by means of acceleration of compressed gas, typically air or oxygen, through a narrow venturi orifice or by means of ultrasonic agitation. 
     Suitable formulations for use in nebulizers consist of the active ingredient in a liquid carrier, the active ingredient comprising up to 40% w/w of the formulation, but preferably less than 20% w/w. The carrier is typically water (and most preferably sterile, pyrogen-free water) or a dilute aqueous alcoholic solution, preferably made isotonic, but may be hypertonic with body fluids by the addition of, for example, sodium chloride. Optional additives include preservatives if the formulation is not made sterile, for example, methyl hydroxybenzoate, antioxidants, flavoring agents, volatile oils, buffering agents and surfactants. 
     Aerosols of solid particles comprising the active compound may likewise be produced with any solid particulate therapeutic aerosol generator. Aerosol generators for administering solid particulate therapeutics to a subject produce particles which are respirable and generate a volume of aerosol containing a predetermined metered dose of a therapeutic at a rate suitable for human administration. One illustrative type of solid particulate aerosol generator is an insufflator. Suitable formulations for administration by insufflation include finely comminuted powders which may be delivered by means of an insufflator or taken into the nasal cavity in the manner of a snuff. In the insufflator, the powder (e.g., a metered dose thereof effective to carry out the treatments described herein) is contained in capsules or cartridges, typically made of gelatin or plastic, which are either pierced or opened in situ and the powder delivered by air drawn through the device upon inhalation or by means of a manually-operated pump. The powder employed in the insufflator consists either solely of the active ingredient or of a powder blend comprising the active ingredient, a suitable powder diluent, such as lactose, and an optional surfactant. The active ingredient typically comprises from 0.1 to 100 w/w of the formulation. 
     A second type of illustrative aerosol generator comprises a metered dose inhaler. Metered dose inhalers are pressurized aerosol dispensers, typically containing a suspension or solution formulation of the active ingredient in a liquefied propellant. During use these devices discharge the formulation through a valve adapted to deliver a metered volume, typically from 10 to 200 ul, to produce a fine particle spray containing the active ingredient. Suitable propellants include certain chlorofluorocarbon compounds, for example, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane and mixtures thereof. The formulation may additionally contain one or more co-solvents, for example, ethanol, surfactants, such as oleic acid or sorbitan trioleate, antioxidant and suitable flavoring agents. 
     An iRNA agent can be incorporated into pharmaceutical compositions suitable for administration. For example, compositions can include one or more species of an iRNA agent and a pharmaceutically acceptable carrier. As used herein the language “pharmaceutically acceptable carrier” is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions. 
     Administration can be provided by the subject or by another person, e.g., a caregiver. A caregiver can be any entity involved with providing care to the human: for example, a hospital, hospice, doctor&#39;s office, outpatient clinic; a healthcare worker such as a doctor, nurse, or other practitioner; or a spouse or guardian, such as a parent. The medication can be provided in measured doses or in a dispenser which delivers a metered dose. 
     The term “therapeutically effective amount” is the amount present in the composition that is needed to provide the desired level of drug in the subject to be treated to give the anticipated physiological response. 
     The term “physiologically effective amount” is that amount delivered to a subject to give the desired palliative or curative effect. 
     The term “pharmaceutically acceptable carrier” means that the carrier can be taken into the lungs with no significant adverse toxicological effects on the lungs. 
     The term “co-administration” refers to administering to a subject two or more agents, and in particular two or more iRNA agents. The agents can be contained in a single pharmaceutical composition and be administered at the same time, or the agents can be contained in separate formulation and administered serially to a subject. So long as the two agents can be detected in the subject at the same time, the two agents are said to be coadministered. 
     The types of pharmaceutical excipients that are useful as carrier include stabilizers such as human serum albumin (HSA), bulking agents such as carbohydrates, amino acids and polypeptides; pH adjusters or buffers; salts such as sodium chloride; and the like. These carriers may be in a crystalline or amorphous form or may be a mixture of the two. 
     Bulking agents that are particularly valuable include compatible carbohydrates, polypeptides, amino acids or combinations thereof. Suitable carbohydrates include monosaccharides such as galactose, D-mannose, sorbose, and the like; disaccharides, such as lactose, trehalose, and the like; cyclodextrins, such as 2-hydroxypropyl-.beta.-cyclodextrin; and polysaccharides, such as raffinose, maltodextrins, dextrans, and the like; alditols, such as mannitol, xylitol, and the like. A preferred group of carbohydrates includes lactose, threhalose, raffinose maltodextrins, and mannitol. Suitable polypeptides include aspartame. Amino acids include alanine and glycine, with glycine being preferred. 
     Suitable pH adjusters or buffers include organic salts prepared from organic acids and bases, such as sodium citrate, sodium ascorbate, and the like; sodium citrate is preferred. 
     Dosage 
     An iRNA agent can be administered at a unit dose less than about 75 mg per kg of bodyweight, or less than about 70, 60, 50, 40, 30, 20, 10, 5, 2, 1, 0.5, 0.1, 0.05, 0.01, 0.005, 0.001, or 0.0005 mg per kg of bodyweight, and less than 200 nmol of iRNA agent (e.g., about 4.4×10 16  copies) per kg of bodyweight, or less than 1500, 750, 300, 150, 75, 15, 7.5, 1.5, 0.75, 0.15, 0.075, 0.015, 0.0075, 0.0015, 0.00075, 0.00015 nmol of iRNA agent per kg of bodyweight. The unit dose, for example, can be administered by injection (e.g., intravenous or intramuscular, intrathecally, or directly into an organ), an inhaled dose, or a topical application. 
     The dosage can be an amount effective to treat or prevent a disease or disorder. It can be given prophylactically or as the primary or a part of a treatment protocol. 
     In one embodiment, the unit dose is administered less frequently than once a day, e.g., less than every 2, 4, 8 or 30 days. In another embodiment, the unit dose is not administered with a frequency (e.g., not a regular frequency). For example, the unit dose may be administered a single time. Because iRNA agent mediated silencing can persist for several days after administering the iRNA agent composition, in many instances, it is possible to administer the composition with a frequency of less than once per day, or, for some instances, only once for the entire therapeutic regimen. 
     In one embodiment, a subject is administered an initial dose, and one or more maintenance doses of an iRNA agent, e.g., a double-stranded iRNA agent, or siRNA agent, (e.g., a precursor, e.g., a larger iRNA agent which can be processed into an siRNA agent, or a DNA which encodes an iRNA agent, e.g., a double-stranded iRNA agent, or siRNA agent, or precursor thereof). The maintenance dose or doses are generally lower than the initial dose, e.g., one-half less of the initial dose. A maintenance regimen can include treating the subject with a dose or doses ranging from 0.01 to 75 mg/kg of body weight per day, e.g., 70, 60, 50, 40, 30, 20, 10, 5, 2, 1, 0.5, 0.1, 0.05, 0.01, 0.005, 0.001, or 0.0005 mg per kg of body weight per day. The maintenance doses are preferably administered no more than once every 5, 10, or 30 days. Further, the treatment regimen may last for a period of time which will vary depending upon the nature of the particular disease, its severity and the overall condition of the patient. In preferred embodiments the dosage may be delivered no more than once per day, e.g., no more than once per 24, 36, 48, or more hours, e.g., no more than once every 5 or 8 days. Following treatment, the patient can be monitored for changes in his condition and for alleviation of the symptoms of the disease state. The dosage of the compound may either be increased in the event the patient does not respond significantly to current dosage levels, or the dose may be decreased if an alleviation of the symptoms of the disease state is observed, if the disease state has been ablated, or if undesired side-effects are observed. 
     The effective dose can be administered in a single dose or in two or more doses, as desired or considered appropriate under the specific circumstances. If desired to facilitate repeated or frequent infusions, implantation of a delivery device, e.g., a pump, semi-permanent stent (e.g., intravenous, intraperitoneal, intracisternal or intracapsular), or reservoir may be advisable. 
     Following successful treatment, it may be desirable to have the patient undergo maintenance therapy to prevent the recurrence of the disease state, wherein the compound of the invention is administered in maintenance doses, ranging from 0.001 g to 100 g per kg of body weight (see U.S. Pat. No. 6,107,094). 
     The concentration of the iRNA agent composition is an amount sufficient to be effective in treating or preventing a disorder or to regulate a physiological condition in humans. The concentration or amount of iRNA agent administered will depend on the parameters determined for the agent and the method of administration, e.g. nasal, buccal, or pulmonary. For example, nasal formulations tend to require much lower concentrations of some ingredients in order to avoid irritation or burning of the nasal passages. It is sometimes desirable to dilute an oral formulation up to 10-100 times in order to provide a suitable nasal formulation. 
     Certain factors may influence the dosage required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and/or age of the subject, and other diseases present. It will also be appreciated that the effective dosage of an iRNA agent such as an siRNA used for treatment may increase or decrease over the course of a particular treatment. Changes in dosage may result and become apparent from the results of diagnostic assays. For example, the subject can be monitored after administering an iRNA agent composition. Based on information from the monitoring, an additional amount of the iRNA agent composition can be administered. 
     Dosing is dependent on severity and responsiveness of the disease condition to be treated, with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of disease state is achieved. Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient. Persons of ordinary skill can easily determine optimum dosages, dosing methodologies and repetition rates. Optimum dosages may vary depending on the relative potency of individual compounds, and can generally be estimated based on EC50s found to be effective in in vitro and in vivo animal models as described above. 
     iRNA agents of the present invention as described herein may be useful in the treatment and (where appropriate) in the prevention of any one of the following diseases/disorders; 
     Cystic fibrosis, Liddles syndrome, renal insufficiency, hypertension, electrolyte imbalances. 
     In particular in some embodiments, iRNA agents of the invention may be used to treat and/or prevent adverse clinical manifestations of these diseases/disorders. 
     The invention is further illustrated by the following examples, which should not be construed as further limiting. 
     EXAMPLES 
     Source of reagents Where the source of a reagent is not specifically given herein, such reagent may be obtained from any supplier of reagents for molecular biology at a quality/purity standard for application in molecular biology. 
     Example 1: Selection of Sequences 
     In order to identify therapeutic siRNAs to downmodulate expression of the alpha subunit of the epithelial sodium channel ENaC (α-ENaC), screening sets were defined based on a bioinformatic analysis. The key drivers for the design of the screening set were predicted specificity of the siRNAs against the transcriptome of the relevant species. For the identification of alpha-ENaC siRNAs and an efficient delivery system a three pronged approach was used: Rat was selected as the test species to address silencing efficacy in vivo after intratracheal delivery, guinea pig was selected as the disease model organism to demonstrate that alpha-ENaC mRNA reduction results in a measurable functional effect. The therapeutic siRNA molecule has to target human alpha-ENaC as well as the alpha-ENaC sequence of at least one toxicology-relevant species, in this case, rhesus monkey. 
     Initial analysis of the relevant alpha-ENaC mRNA sequence revealed few sequences can be identified that fulfil the specificity requirements and at the same time target alpha-ENaC mRNA in all relevant species. Therefore it was decided to design independent screening sets for the therapeutic siRNA and for the surrogate molecules to be tested in the relevant disease model (Tables 1A, 1B, 1C and 1D). 
     All siRNAs recognize the human alpha-ENaC sequence, as a human cell culture system was selected for determination of in vitro activity (H441, see below). Therefore all siRNAs can be used to target human alpha-ENaC mRNA in a therapeutic setting. 
     The therapeutic screening sets were designed to contain only siRNA sequences that are fully complementary to the human and rhesus monkey alpha-ENaC sequences. 
     Design and in silico selection of siRNAs targeting alpha-ENaC (SCNN1A) siRNA design was carried out to identify siRNAs for the four previously defined sets (see above)
         a) “Initial screening set”   b) “Extended screening set”   c) “In vivo surrogate set for rat”   d) “In vivo surrogate set for guinea pig”       

     Initial screening set The aim for in silico selection of an initial screening set was to identify siRNAs specifically targeting human alpha-ENaC, as well as its rhesus monkey ortholog. The human target mRNA (NM_001038.4) was downloaded from NCBI resource (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?CMD=search&amp;DB=nucleotide) during the complete siRNA selection procedure. In order to identify the alpha-ENaC rhesus ( Macaca mulatta ) ortholog, the human sequence was used in a blastn search at Baylor College of Medicine (http://www.hgsc.bcm.tmc.edu/blast/?organism=Mmulatta) against Mmulatta contigs as of 2004 10 01. All hit regions were extracted and assembled by the CAP assembly tool to generate a first assembly sequence. Further, a BLAST search was performed with the human sequence at UCSC (http://genome.ucsc.edu/cgi-bin/hgBlat?command=start&amp;org=Rhesus&amp;db=rheMac2&amp;hgsid=84859356) against Rhesus freeze 12 Mar. 2005. The scaffold hit 84554 was downloaded and used together with the first assembly sequence by CAP to generate the final consensus sequence for rhesus alpha-ENaC. 
     Following extraction of all overlapping 19mer sequences out of the human mRNA, conserved 19mers were identified that had identical sequences in the assembled rhesus consensus sequence. Those 19mer sequences were defined as the pool of human-rhesus cross-reactive siRNA (sense) sequences, represented by 1185 19mers. 
     The corresponding antisense sequences were generated and tested for specificity in human. For this, their predicted potential for interacting with irrelevant target mRNAs (off-target potential) was taken as parameter. Sequences with low off-target potential were defined as preferable and predicted to be more specific. 
     For further selection, candidate siRNAs were ranked according to their predicted potential for interacting with other host sequences (here, without limitation, human). siRNAs with low off-target potential are assumed to be more specific in vivo. For predicting siRNA-specific off-target potential, the following assumptions were made:
         1) off-target potential of a strand can be deduced from the number and distribution of mismatches to an off-target   2) the most relevant off-target, that is the gene predicted to have the highest probability to be silenced due to tolerance of mismatches, determines the off-target potential of the strand   3) positions 2 to 9 (counting 5′ to 3′) of a strand (seed region) may contribute more to off-target potential than rest of sequence (that is non-seed and cleavage site region) (Haley, B., and Zamore, P. D., Nat Struct Mol Biol. 2004, 11:599).   4) positions 10 and 11 (counting 5′ to 3′) of a strand (cleavage site region) may contribute more to off-target potential than non-seed region (that is positions 12 to 18, counting 5′ to 3′)   5) positions 1 and 19 of each strand are not relevant for off-target interactions   6) off-target potential can be expressed by the off-target score of the most relevant off-target, calculated based on number and position of mismatches of the strand to the most homologous region in the off-target gene considering assumptions 3 to 5   7) assuming potential abortion of sense strand activity by internal modifications introduced, only off-target potential of antisense strand will be relevant       

     To identify potential off-target genes, 19mer antisense sequences were subjected to a homology search against publicly available human mRNA sequences, assumed to represent the human comprehensive transcriptome. 
     To this purpose, fastA (version 3.4) searches were performed with all 19mer sequences against a human RefSeq database (available version from ftp://ftp.ncbi.nih.gov/refseq/ on Nov. 18, 2005). FastA search was executed with parameters-values-pairs -f 30 -g 30 in order to take into account the homology over the full length of the 19mer without any gaps. In addition, in order to ensure the listing of all relevant off-target hits in the fastA output file the parameter -E 15000 was used. 
     The search resulted in a list of potential off-targets for each input sequence listed by descending sequence homology over the complete 19mer. 
     To rank all potential off-targets according to assumptions 3 to 5, and by this identify the most relevant off-target gene and its off-target score, fastA output files were analyzed by a perl script. 
     The script extracted the following off-target properties for each 19mer input sequence and each off-target gene to calculate the off-target score:
         Number of mismatches in non-seed region   Number of mismatches in seed region   Number of mismatches in cleavage site region
 
The off-target score was calculated by considering assumptions 3 to 5 as follows:
       

     
       
         
           
             
               
                 
                   
                     Off 
                     ⁢ 
                     
                       - 
                     
                     ⁢ 
                     target 
                     ⁢ 
                     
                         
                     
                     ⁢ 
                     score 
                   
                   = 
                       
                   ⁢ 
                   
                     number 
                     ⁢ 
                     
                         
                     
                     ⁢ 
                     of 
                     ⁢ 
                     
                         
                     
                     ⁢ 
                     seed 
                     ⁢ 
                     
                         
                     
                     ⁢ 
                     
                       mismatches 
                       * 
                     
                     ⁢ 
                     10 
                   
                 
               
             
             
               
                 
                   
                     + 
                         
                     ⁢ 
                     number 
                   
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   of 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   cleavage 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   site 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   
                     mismatches 
                     * 
                   
                   ⁢ 
                   1.2 
                 
               
             
             
               
                 
                   
                     + 
                         
                     ⁢ 
                     number 
                   
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   of 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   non 
                   ⁢ 
                   
                     - 
                   
                   ⁢ 
                   seed 
                   ⁢ 
                   
                       
                   
                   ⁢ 
                   
                     mismatches 
                     * 
                   
                   ⁢ 
                   1 
                 
               
             
           
         
       
     
     The most relevant off-target gene for each 19mer sequence was defined as the gene with the lowest off-target score. Accordingly, the lowest off-target score was defined as representative for the off-target potential of each siRNA, represented by the 19mer antisense sequence analyzed. 
     Calculated off-target potential was used as sorting parameter (descending by off-target score) in order to generate a ranking for all human-rhesus cross-reactive siRNA sequences. 
     An off-target score of 3 or more was defined as prerequisite for siRNA selection, whereas all sequences containing 4 or more G&#39;s in a row (poly-G sequences) were excluded, leading to selection of a total of 152 siRNAs targeting human and rhesus ENaC alpha (see Table 1a). 
     Extended screening set The aim for in silico selection of the extended screening set was to identify all further siRNAs targeting human alpha-ENaC with sufficient specificity, that were excluded from the initial set due to missing cross-reactivity to rhesus. The remaining sequences from the pool of 19mers derived from human alpha-ENaC that have not been analyzed before were taken and the corresponding antisense sequences were generated. The most relevant off-target gene and its corresponding off-target scores were calculated as described in section “Initial screening set”. 
     For determining cross-reactivity to mouse and guinea pig ( Cavia porcellus /cobya), alpha-ENaC sequences of these species were downloaded from NCBI nucleotide database 1  (accession numbers NM_011324.1 and AF071230 (full length)/DQ109811 (partial cds), respectively). The two guinea pig sequences were used to generate a guinea pig alpha-ENaC consensus sequence. Every human 19mer sequence was tested for presence in the mouse and guinea pig sequences. Positive sequences were assigned to the pool of human-mouse cross-reactive siRNA (sense) sequences, or human-guinea pig cross-reactive siRNA (sense) sequences. After exclusion of all poly-G sequences, sequences were selected with off-target scores of 3 or more as well as those with off-target scores of 2.2 or 2.4 and cross-reactivity to mouse, rhesus or guinea pig. The total number of siRNAs in the extended screening pool was 344 (see Table 1b). 
     In vivo rat surrogate set The aim for in silico selection of the in vivo rat surrogate set was to identify all siRNAs targeting human and rat alpha-ENaC with sufficient specificity in rat. For identification of human-rat cross-reactive siRNAs, rat alpha-ENaC mRNA sequence was downloaded from NCBI nucleotide database (accession number, NM_031548.2), and all sequences out of the pool of human 19mers were tested for presence in the rat sequence, representing the pool of human-rat cross-reactive siRNA (sense) sequences. 
     The corresponding antisense sequences were generated and tested for specificity in rat. For this, the most relevant off-target gene in rat and its corresponding off-target scores were calculated as described in section “Initial screening set” using the rat mRNA set (RefSeq database) instead of the human transcripts. After exclusion of all poly-G sequences, a ranking was generated considering the rat off-target score in first priority and the human off-target score with second priority. Those 48 sequences from the top of the list were finally selected representing the in vivo rat surrogate set (see Table 1c). 
     In vivo guinea pig surrogate set The aim for in silico selection of the in vivo guinea pig surrogate set was to identify all siRNAs targeting human and guinea pig alpha-ENaC that have not been selected in previous sets. The remaining siRNAs of the previously determined set of human-guinea pig cross-reactive siRNA (sense) sequences were ranked according to human off-target scores. The top 63 sequences (excluding poly-G sequences) were selected, representing the in vivo guinea pig surrogate set (see Table 1d). 
     Example 2: siRNA Synthesis 
     Synthesis of nucleotides comprising natural bases. As the siRNAs from the screening sets are all potentially intended for in vivo administration, siRNAs were synthesised with a modification strategy that protects the siRNAs from degradation by endo- and exonucleases in a biological environment. In this strategy, the 3′-ends of both strands are protected from a 3′-&gt;5′-exonucleotitic activity by a phosphorothioate linkage between the two last nucleobases at the 3′-end. In order to inhibit endo-nucleolytic degradation of the siRNA all pyrimidines in the sense strand of the siRNA were replaced with the corresponding 2′-O-methyl-modified ribonucleotide. To reduce the number of modifications in the antisense strand, which is the more active strand and therefore more sensitive to modifications, we only modified the pyrimidines in the context of previously identified major nuclease cleavage sites with 2′-O-methyl groups. The major cleavage sites are the following two sequence motifs: 5′-UA-3′ and 5′-CA-3′. 
     Since it has also been considered to use siRNAs in formulations that potentially protect the RNAs from the nucleolytic biological environment in the lung, the same set of siRNAs were also synthesized without any protection from endonucleolytic degradation. 
     Where the source of a reagent is not specifically given herein, such reagent may be obtained from any supplier of reagents for molecular biology at a quality/purity standard for application in molecular biology. 
     Single-stranded RNAs were produced by solid phase synthesis on a scale of 1 μmole using an Expedite 8909 synthesizer (Applied Biosystems, Applera Deutschland GmbH, Darmstadt, Germany) and controlled pore glass (CPG, 500 Å, Proligo Biochemie GmbH, Hamburg, Germany) as solid support. RNA and RNA containing 2′-O-methyl nucleotides were generated by solid phase synthesis employing the corresponding phosphoramidites and 2′-O-methyl phosphoramidites, respectively (Proligo Biochemie GmbH, Hamburg, Germany). These building blocks were incorporated at selected sites within the sequence of the oligoribonucleotide chain using standard nucleoside phosphoramidite chemistry such as described in Current protocols in nucleic acid chemistry, Beaucage, S. L. et al. (Edrs.), John Wiley &amp; Sons, Inc., New York, N.Y., USA. Phosphorothioate linkages were introduced by replacement of the iodine oxidizer solution with a solution of the Beaucage reagent (Chruachem Ltd, Glasgow, UK) in acetonitrile (1%). Further ancillary reagents were obtained from Mallinckrodt Baker (Griesheim, Germany). 
     Deprotection and purification of the crude oligoribonucleotides by anion exchange HPLC were carried out according to established procedures. Yields and concentrations were determined by UV absorption of a solution of the respective RNA at a wavelength of 260 nm using a spectral photometer (DU 640B, Beckman Coulter GmbH, Unterschleißheim, Germany). Double stranded RNA was generated by mixing an equimolar solution of complementary strands in annealing buffer (20 mM sodium phosphate, pH 6.8; 100 mM sodium chloride), heated in a water bath at 85-90° C. for 3 minutes and cooled to room temperature over a period of 3-4 hours. The annealed RNA solution was diluted to a concentration of 50 μmole double stranded RNA/L and stored at −20° C. until use. 
     Example 3: siRNA Testing In Vitro 
     The ability of the iRNA agents to inhibit expression of alpha-ENaC was tested in human cell lines in vitro, or in rats in vivo. The iRNA agent is transfected into the cells, e.g., by transfection, allowed to act on the cells for a certain time, e.g., 24 hours, and levels of alpha-ENaC mRNA were determined by branched-DNA analysis. Alternatively, the iRNA agent is administered in vivo via the intratracheal route and the inhibition of alpha-ENaC mRNA expression determined by branched-DNA analysis on the target organ. Complementing these direct assays, we tested the inhibition of target gene expression by RNAi agents for alpha-ENaC mRNA recombinantly expressed in mammalian host cells. 
     Cell lines. H441 cells were obtained from the American Type Culture Collection (ATCC-Number: HTB-174, LCG Promochem GmbH, Wesel, Germany) and were grown in RPMI 1640, 10% fetal calf serum, 100u penicillin/100 μg/mL streptomycin, 2 mM L-glutamine, 10 nM Hepes and 1 mM Sodium-Pyruvate (all from Biochrom AG, Berlin, Germany) at 37° C. under a 5% CO 2 /95% air atmosphere. 
     Primary human bronchial epithelial cells were obtained from Cambrex (Cat #CC-2540) and were routinely grown in BEGM media with singlequots (Cambrex Cat #CC-3170 minus tri-iodothreonine). For polarisation and growth at air liquid interface the cells were grown in a 1:1 mixture of BEGM:DMEM supplemented with 4.5 g/L D-Glucose (Gibco BRL Cat #41965-039) and supplemented with singlequots (Cambrex Cat #CC-4175), as above but minus the tri-iodothreonine and GA1000 aliquots and in the presence of 50 μg/mL Gentamycin (Gibco Brl Cat #10131-015). As cells were maintained in serum-free media, trypsin neutralisation solution was used during passaging steps (Cambrex Cat #CC-5002). For polarisation and culture at air-liquid interface the cells were grown on semipermeable (0.4 micron) polycarbonate supports (Corning Costar Cat #3407 #3460) and cultured throughout at 37° C. under a 5% CO 2 /95% air atmosphere. 
     Cos-1 African green monkey kidney cells (ATCC #CRL-1650) were grown in Dulbecco&#39;s MEM, 4.5 g/L glucose, 10% fetal bovine serum, 2 mM L-glutamine, 1.5 g/L sodium bicarbonate (Gibco BRL), 100u penicillin/100 μg/mL streptomycin. 
     Example 3.1: In Vitro Screen for Active Alpha-ENaC siRNAs and IC50 Determination in H441 
     One day prior to transfection, ENaC-alpha expression was induced in H441 cells (ATCC-Number: HTB-174, LCG Promochem GmbH, Wesel, Germany) by adding 100 nM of dexamthasone. Directly before transfection, cells were seeded at 1.5×10 4  cells/well on 96-well plates (Greiner Bio-One GmbH, Frickenhausen, Germany) in 75 μL of growth medium (RPMI 1640, 10% fetal calf serum, 100u penicillin/100 μg/ml streptomycin, 2 mM L-glutamine, 10 nM Hepes and 1 mM Sodium-Pyruvate, all from Biochrom AG, Berlin, Germany). Transfections were performed in quadruplicates. For each well 0.5 μL Lipofectamine2000 (Invitrogen GmbH, Karlsruhe, Germany) were mixed with 12 μL Opti-MEM (Invitrogen) and incubated for 15 min at room temperature. For the siRNA concentration being 50 nM in the 100 μL transfection volume, 1 μL of a 5 μM siRNA were mixed with 11.5 μL. Opti-MEM per well, combined with the Lipofectamine2000-Opti-MEM mixture and again incubated for 15 minutes at room temperature. siRNA-Lipofectamine2000-complexes were applied completely (25 μL each per well) to the cells and cells were incubated for 24 h at 37° C. and 5% CO 2  in a humidified incubator (Heraeus GmbH, Hanau). 
     Cells were harvested by applying 50 μL of lysis mixture (content of the QuantiGene bDNA-kit from Genospectra, Fremont, USA) to each well containing 100 μL of growth medium and were lysed at 53° C. for 30 min. Afterwards, 50 μL of the lysates were incubated with probesets specific to human ENaC-alpha and human GAPDH (sequence of probesets see below) and proceeded according to the manufacturer&#39;s protocol for QuantiGene. In the end chemoluminescence was measured in a Victor2-Light (Perkin Elmer, Wiesbaden, Germany) as RLUs (relative light units) and values obtained with the hENaC probeset were normalized to the respective GAPDH values for each well. Values obtained with siRNAs directed against ENaC-alpha were related to the value obtained with an unspecific siRNA (directed against HCV) which was set to 100%. The percentage residual expression of alpha-ENaC for siRNA examples is shown in Tables 1A-1D. 
     Effective siRNAs from the screen were further characterized by dose response curves. Transfections of dose response curves were performed at the following concentrations: 100 nM, 16.7 nM, 2.8 nM, 0.46 nM, 77 picoM, 12.8 picoM, 2.1 picoM, 0.35 picoM, 59.5 fM, 9.9 fM and mock (no siRNA) and diluted with Opti-MEM to a final concentration of 12.5 μl according to the above protocol. Data analysis was performed using Microsoft Excel add-in software XL-fit 4.2 (IDBS, Guildford, Surrey, UK) and applying the sigmoidal model number 606. 
     Probesets: 
     Human Alpha-ENaC: 
       
     
       
         
           
               
               
               
               
               
             
               
                   
               
               
                   
                   
                   
                   
                 SEQ. ID. 
               
               
                 FPL Name 
                 Members 
                 Function 
                 Sequence 
                 NO: 
               
               
                   
               
             
            
               
                 hENAC001 
                 .235.255.CE 
                 CE 
                 gtctgtccagggtttccttccTTTTTctcttggaaagaaag 
                 1645 
               
               
                   
                   
                   
                 t 
                   
               
               
                   
               
               
                 hENAC002 
                 .274.293.CE 
                 CE 
                 actgccattcttggtgcagtTTTTTctcttggaaagaaagt 
                 1646 
               
               
                   
               
               
                 hENAC003 
                 .344.367.CE 
                 CE 
                 ctctcctggaagcaggagtgaataTTTTTctcttggaaaga 
                 1647 
               
               
                   
                   
                   
                 aagt 
                   
               
               
                   
               
               
                 hENAC004 
                 .391.411.CE 
                 CE 
                 gccgcggatagaagatgtaggTTTTTctcttggaaagaaag 
                 1648 
               
               
                   
                   
                   
                 t 
                   
               
               
                   
               
               
                 hENAC005 
                 .501.521.CE 
                 CE 
                 gcacttggtgaaacagcccagTTTTTctcttggaaagaaag 
                 1649 
               
               
                   
                   
                   
                 t 
                   
               
               
                   
               
               
                 hENAC006 
                 .539.560.CE 
                 CE 
                 agcagagagctggtagctggtcTTTTTctcttggaaagaaa 
                 1650 
               
               
                   
                   
                   
                 gt 
                   
               
               
                   
               
               
                 hENAC007 
                 .256.273.LE 
                 LE 
                 cgccataatcgcccccaaTTTTTaggcataggacccgtgtc 
                 1651 
               
               
                   
                   
                   
                 t 
                   
               
               
                   
               
               
                 hENAC008 
                 .368.390.LE 
                 LE 
                 cacagccacactccttgatcatgTTTTTaggcataggaccc 
                 1652 
               
               
                   
                   
                   
                 gtgtct 
                   
               
               
                   
               
               
                 hENAC009 
                 .412.431.LE 
                 LE 
                 acagtactccacgttctgggTTTTTaggcataggacccgtg 
                 1653 
               
               
                   
                   
                   
                 tct 
                   
               
               
                   
               
               
                 hENAC010 
                 .455.477.LE 
                 LE 
                 ggagcttatagtagcagtaccccTTTTTaggcataggaccc 
                 1654 
               
               
                   
                   
                   
                 gtgtct 
                   
               
               
                   
               
               
                 hENAC011 
                 .522.538.LE 
                 LE 
                 acgctgcatggcttccgTTTTTaggcataggacccgtgtct 
                 1655 
               
               
                   
               
               
                 hENAC012 
                 .561.580.LE 
                 LE 
                 gagggccatcgtgagtaaccTTTTTaggcataggacccgtg 
                 1656 
               
               
                   
                   
                   
                 tct 
                   
               
               
                   
               
               
                 hENAC013 
                 .214.234.BL 
                 BL 
                 Tcatgctgatggaggtctcca 
                 1657 
               
               
                   
               
               
                 hENAC014 
                 .294.318.BL 
                 BL 
                 Ggtaaaggttctcaacaggaacatc 
                 1658 
               
               
                   
               
               
                 hENAC015 
                 .319.343.BL 
                 BL 
                 Cacacctgctgtgtgtactttgaag 
                 1659 
               
               
                   
               
               
                 hENAC016 
                 .432.454.BL 
                 BL 
                 Caggaactgtgcffictgtagtc 
                 1660 
               
               
                   
               
               
                 hENAC017 
                 .478.500.BL 
                 BL 
                 Gtggtctgaggagaagtcaacct 
                 1661 
               
               
                   
               
               
                 hENAC018 
                 .581.599.BL 
                 BL 
                 Ccattcctgggatgtcacc 
                 1662 
               
               
                   
               
            
           
         
       
     
     Human GAPDH: 
       
     
       
         
           
               
               
               
               
               
             
               
                   
               
               
                 FPL 
                   
                   
                   
                 SEQ. ID. 
               
               
                 Name 
                 Members 
                 Function 
                 Sequence 
                 NO: 
               
               
                   
               
             
            
               
                 hGAP001 
                 AF261085.252.271.CE 
                 CE 
                 gaatttgccatgggtggaatTTTTTctcttggaaagaaag 
                 1663 
               
               
                   
                   
                   
                 t 
                   
               
               
                   
               
               
                 hGAP002 
                 AF261085.333.352.CE 
                 CE 
                 ggagggatctcgctcctggaTTTTTctcttggaaagaaag 
                 1664 
               
               
                   
                   
                   
                 t 
                   
               
               
                   
               
               
                 hGAP003 
                 AF261085.413.431.CE 
                 CE 
                 ccccagccttctccatggtTTTTTctcttggaaagaaagt 
                 1665 
               
               
                   
               
               
                 hGAP004 
                 AF261085.432.450.CE 
                 CE 
                 gctcccccctgcaaatgagTTTTTctcttggaaagaaagt 
                 1666 
               
               
                   
               
               
                 hGAP005 
                 AF261085.272.289.LE 
                 LE 
                 agccttgacggtgccatgTTTTTaggcataggacccgtgt 
                 1667 
               
               
                   
                   
                   
                 ct 
                   
               
               
                   
               
               
                 hGAP006 
                 AF261085.290.310.LE 
                 LE 
                 gatgacaagcttcccgttctcTTTTTaggcataggacccg 
                 1668 
               
               
                   
                   
                   
                 tgtct 
                   
               
               
                   
               
               
                 hGAP007 
                 AF261085.311.332.LE 
                 LE 
                 agatggtgatgggatttccattTTTTTaggcataggaccc 
                 1669 
               
               
                   
                   
                   
                 gtgtct 
                   
               
               
                   
               
               
                 hGAP008 
                 AF261085.353.372.LE 
                 LE 
                 gcatcgccccacttgattttTTTTTaggcataggacccgt 
                 1670 
               
               
                   
                   
                   
                 gtct 
                   
               
               
                   
               
               
                 hGAP009 
                 AF261085.373.391.LE 
                 LE 
                 cacgacgtactcagcgccaTTTTTaggcataggacccgtg 
                 1671 
               
               
                   
                   
                   
                 tct 
                   
               
               
                   
               
               
                 hGAP010 
                 AF261085.451.472.LE 
                 LE 
                 ggcagagatgatgacccttttgTTTTTaggcataggaccc 
                 1672 
               
               
                   
                   
                   
                 gtgtct 
                   
               
               
                   
               
               
                 hGAP011 
                 AF261085.392.412.BL 
                 BL 
                 Ggtgaagacgccagtggactc 
                 1673 
               
               
                   
               
            
           
         
       
     
     The IC 50 s for siRNA examples is shown in Table 2A and 2B. 
     Example 3.2: Transient Alpha-ENaC Knockdown in a Primary Human Bronchial Epithelial Model 
     Human bronchial epithelial cells (donor reference 4F1499) were plated in 24-well plates at 1×10 5  cells per well in 0.5 mL growth medium one day before transfection. The cells were 70% confluent on the day of siRNA transfection. 
     Each siRNA was resuspended at 100 nM in 1 mL of Optimem I (Invitrogen) and in a separate tube, Lipofectamine 2000 (Invitrogen) was diluted to 6 μL/mL in Optimem, giving an amount sufficient for transfection of four replicates in a 24-well plate. After 5 minutes at room temperature, the mixtures were combined to give the desired final concentration of 50 nM siRNA and 3 μL/mL Lipofectamine 2000. The transfection mixture was incubated for a further 20 minutes at room temperature and 420 μL of the siRNA/reagent complex was added to each well as dictated by the experimental design. Plates were gently rocked to ensure complete mixing and then incubated at 37° C. in an incubator at 5% CO 2 /95% air for 4 hours. Subsequently, the transfection mixture was aspirated and the cells were returned to normal culture conditions for a further 20 hours. 
     Cell lysates were prepared for branched-DNA analysis. A 2:1 medium:lysis buffer (Panomics) mixture was prepared and cells were lysed in 200 μL at 53° C. for 30 minutes. After a visual check for complete lysis, the cell lysates were stored at −80° C. for subsequent analysis. Branched-DNA analysis was performed as described above, with alpha-ENaC expression normalized against GAPDH. The branched DNA analysis protocol used differs from that above only in that 20 μL of sample was applied to each well in this case. 
     Table 2C shows the alpha-ENaC expression in primary HBEC for siRNA examples. 
     Example 3.3: In Vitro Inhibition of Exogenously Expressed Cloned Cynomolgous Alpha-ENaC Gene Expression for Selected RNAi Agents in Cos-1 Cells 
     Cloning of the cynomolgous alpha-ENaC sequence. Primer sequences for amplification of 5′-UTR and CDS (nucleotides shown in brackets correspond to the  Macaca mulatta  (Rhesus monkey) α-ENaC cDNA sequence): 
     
       
         
           
               
               
               
            
               
                   
                   
                 (nt 1427) 
               
               
                   
                   
                 P745: 
               
            
           
           
               
               
            
               
                   
                 (SEQ. ID. NO: 1674) 
               
            
           
           
               
               
               
            
               
                   
                   
                 5′-CTCCATGTTCTGCGGCCGCGGATAGAAG-3′ 
               
               
                   
                   
               
               
                   
                   
                 (nt 1) 
               
               
                   
                   
                 P733: 
               
            
           
           
               
               
            
               
                   
                 (SEQ. ID. NO: 1675) 
               
            
           
           
               
               
               
            
               
                   
                   
                 5′-CCGGCCGGCGGGCGGGCT-3′ 
               
               
                   
                   
               
               
                   
                   
                 (nt 17) 
               
               
                   
                   
                 P734: 
               
            
           
           
               
               
            
               
                   
                 (SEQ. ID. NO: 1676) 
               
            
           
           
               
               
               
            
               
                   
                   
                 5-CTCCCCAGCCCGGCCGCT-3′ 
               
               
                   
                   
               
               
                   
                   
                 (nt 28) 
               
               
                   
                   
                 P735: 
               
            
           
           
               
               
            
               
                   
                 (SEQ. ID. NO: 1677) 
               
            
           
           
               
               
               
            
               
                   
                   
                 5′-GGCCGCTGCACCTGTAGGG-3′ 
               
               
                   
                   
               
               
                   
                   
                 Primer sequences for amplification 
               
               
                   
                   
                 of CDS and 3′-UTR: 
               
               
                   
                   
                 (nt 1422) 
               
               
                   
                   
                 P737: 
               
            
           
           
               
               
            
               
                   
                 (SEQ. ID. NO: 1678) 
               
            
           
           
               
               
               
            
               
                   
                   
                 5′-ATGGAGTACTGTGACTACAGG-3′ 
               
               
                   
                   
               
               
                   
                   
                 (nt 3113) 
               
               
                   
                   
                 P740: 
               
            
           
           
               
               
            
               
                   
                 (SEQ. ID. NO: 1679) 
               
            
           
           
               
               
               
            
               
                   
                   
                 5′-TTGAGCATCTGCCTACTTG-3′ 
               
               
                   
                   
               
               
                   
                   
                 Primer sequences for amplification 
               
               
                   
                   
                 of internal part of CDS: 
               
               
                   
                   
                 (nt 1182) 
               
               
                   
                   
                 P713: 
               
            
           
           
               
               
            
               
                   
                 (SEQ. ID. NO: 1679) 
               
            
           
           
               
               
               
            
               
                   
                   
                 5′-5′-ATGGATGATGGTGGCTTTAACTTGCGG-3′ 
               
               
                   
                   
               
               
                   
                   
                 (nt 2108) 
               
               
                   
                   
                 P715: 
               
            
           
           
               
               
            
               
                   
                 (SEQ. ID. NO: 1680) 
               
            
           
           
               
               
               
            
               
                   
                   
                 5′-5′-TCAGGGCCCCCCCAGAGG-3′ 
               
            
           
         
       
     
     Cynomolgus ( Macaca fascicularis ) lung total RNA (#R1534152-Cy-BC) was purchased from BioCat (Germany). Synthesis of cDNA was performed using the SuperScript III First Strand Synthesis System (Invitrogen). Synthesis of cDNA was performed using either random hexamers or oligo dT primers. In addition, cynomolgus lung first strand cDNA was also purchased from BioCat/#C1534160-Cy-BC). For PCR amplification, the Advantage 2 PCR kit (#K1910-1, Clontech) was used. Amplification of the 5′-UTR and parts of the CDS was performed using P745 and a equimolar mixture of P733, P734 and P735. For PCR amplification of the CDS and 3′-UTR, primers P737 and P740 were used. The primers P713 and P715 were used for amplification of parts of the CDS. 
     All PCR products were analysed by agarose gel electrophoresis and then cloned into the pCR2.1 vector using the TOPO TA cloning kit (Invitrogen) in TOP10 bacteria. Clones were then picked and DNA was isolated using the Qiagen Miniprep kit. After restriction enzyme digest with EcoRI and analysis by agarose gel electrophoresis, DNA from correct clones were subjected to sequencing. 
     The sequences were then aligned with the α-ENaC cDNA sequence of Rhesus monkey, and sequences of the individual clones were aligned with each other. The full-length cynomolgus alpha-ENaC cDNA was then cloned by digestion of the 5′-part (5′-UTR and CDS, clone 55) with EcoR I and Not I, digestion of the middle part of the CDS by Not I and BstE II (clone 15), and the 3′-part (CDS and 3′-UTR) by BstE II and EcoR V (clone 80). The digested DNA fragments were then subcloned into pcDNA3.1, digested with EcoR I and EcoR V. The full-length cynomolgus alpha-ENaC cDNA in pcDNA3.1 was then subjected to full-length sequencing (Ingenetix, Vienna, Austria). The cynomolgus alpha-ENaC cDNA sequence corresponds to nt 28-3113 of the Rhesus alpha-ENaC cDNA sequence. Finally the cynomolgus alpha-ENaC cDNA was then excised from pcDNA3.1-cynomolgus alpha-ENaC by digestion with BamH I and EcoR V and subcloned into the vector pXOON. The plasmid map is illustrated in  FIG. 1 .  FIG. 2  depicts the protein (SEQ.I.D.NO:1681) and DNA (SEQ.I.D.NO:1682) sequence of cynomolgous alpha-ENaC. 
     Transfections: COS-1 cells were seeded at 6×10 4  cells/well on 24 well plates each in 0.5 mL of growth medium. One day after seeding the cells were co-transfected with the pXOON cynomolgous alpha-ENaC expression plasmid and the indicated siRNA. For each well, 4 ng of alpha-ENaC expression plasmid and 600 ng carrier plasmid (pNFAT-luc) were co-transfected with the relevant siRNA (final concentration 45 nM) using X-treme gene transfection reagent (Roche) at 3.75 μL/well in a total volume of 7204/well Opti-MEM (Invitrogen) as described below. 
     Transfections were performed in triplicate for each sample. Plasmid/siRNA mastermixes (each for 3.5 wells) were prepared as follows: 14 ng alpha-ENaC expression plasmid, 2.1 μg carrier plasmid and 112 pmoles of relevant siRNA in a total volume 210 μL (Opti-MEM). A lipid mastermix was prepared for the whole transfection (105 μL lipid plus 15754 Opti-MEM for eight triplicate transfection samples). Plasmid/siRNA and lipid were mixed in equal volume to give a total volume of 420 μL transfection mix per triplicate sample (3.5×). Following a 20 minute incubation at room temperature, 120 μL of the relevant transfection mix was added to each well of cells in a final transfection volume of 720 μL (Opti-MEM). Cells were transfected for 24 hours at 37° C. and 5% CO 2  in a humidified incubator (Heraeus GmbH, Hanau, Germany) and harvested for branched-DNA analysis. 
     Cell lysates were prepared for branched DNA analysis. A 2:1 medium:lysis buffer (Panomics) mixture was prepared and cells were lysed in 200 μL at 53° C. for 30 minutes. After a visual check for complete lysis, the cell lysates were stored at −80° C. for subsequent analysis. Branched-DNA analysis was performed as described above, with cyno alpha-ENaC expression normalized against eGFP from the expression plasmid. The branched-DNA analysis protocol used differs from that above only in that 20 μL of sample was applied to each well in this case. 
     Probesets: Cynomolgous Alpha-ENaC: 
     
       
         
           
               
               
               
               
             
               
                   
               
               
                 FPL Name 
                 Function 
                 Sequence 
                 SEQ ID NO: 
               
               
                   
               
             
            
               
                 cyENa001 
                 CE 
                 cgccgtgggctgctgggTTTTTctcttggaaagaaagt 
                 1683 
               
               
                   
               
               
                 cyENa002 
                 CE 
                 ggtaggagcggtggaactcTTTTTctcttggaaagaaagt 
                 1684 
               
               
                   
               
               
                 cyENa003 
                 CE 
                 cagaagaactcgaagagctctcTTTTTctcttggaaagaaagt 
                 1685 
               
               
                   
               
               
                 cyENa004 
                 CE 
                 cccagaaggccgtcttcatTTTTTctcttggaaagaaagt 
                 1686 
               
               
                   
               
               
                 cyENa005 
                 LE 
                 ggtgcagagccagagcactgTTTTTctcttggaaagaaagt 
                 1687 
               
               
                   
               
               
                 cyENa006 
                 LE 
                 gtgccgcaggttctgggTTTTTaggcataggacccgtgtct 
                 1688 
               
               
                   
               
               
                 cyENa007 
                 LE 
                 gatcagggcctcctcctcTTTTTaggcataggacccgtgtct 
                 1689 
               
               
                   
               
               
                 cyENa008 
                 LE 
                 ccgtggatggtggtattgttgTTTTTaggcataggacccgtgtct 
                 1690 
               
               
                   
               
               
                 cyENa009 
                 LE 
                 gcggttgtgctgggagcTTTTTaggcataggacccgtgtct 
                 1691 
               
               
                   
               
               
                 cyENa0010 
                 LE 
                 ttgccagtacatcatgccaaaTTTTTaggcataggacccgtgtct 
                 1692 
               
               
                   
               
               
                 cyENa0011 
                 BL 
                 acaccaggcggatggcg 
                 1693 
               
               
                   
               
            
           
         
       
     
     eGFP: 
     
       
         
           
               
               
               
               
             
               
                   
               
               
                 FPL Name 
                 Function 
                 Sequence 
                 SEQ ID NO: 
               
               
                   
               
             
            
               
                 EGFP001 
                 CE 
                 ggcacgggcagcttgcTTTTTctcttggaaagaaagt 
                 1694 
               
               
                   
               
               
                 EGFP002 
                 CE 
                 ggtagcggctgaagcactgTTTTTctcttggaaagaaagt 
                 1695 
               
               
                   
               
               
                 EGFP003 
                 CE 
                 cctggacgtagccttcgggTTTTTctcttggaaagaaagt 
                 1696 
               
               
                   
               
               
                 EGFP004 
                 CE 
                 ccttgaagaagatggtgcgctTTTTTctcttggaaagaaagt 
                 1697 
               
               
                   
               
               
                 EGFP005 
                 LE 
                 cgaacttcacctcggcgcTTTTTctcttggaaagaaagt 
                 1698 
               
               
                   
               
               
                 EGFP006 
                 LE 
                 ccttcagctcgatgcggtTTTTTctcttggaaagaaagt 
                 1699 
               
               
                   
               
               
                 EGFP007 
                 LE 
                 gtcacgagggtgggccagTTTTTaggcataggacccgtgtct 
                 1700 
               
               
                   
               
               
                 EGFP008 
                 LE 
                 cacgccgtaggtcagggtgTTTTTaggcataggacccgtgtct 
                 1701 
               
               
                   
               
               
                 EGFP009 
                 LE 
                 gtgctgcttcatgtggtcggTTTTTaggcataggacccgtgtct 
                 1702 
               
               
                   
               
               
                 EGFP0010 
                 LE 
                 tcaccagggtgtcgccctTTTTTaggcataggacccgtgtct 
                 1703 
               
               
                   
               
               
                 EGFP0011 
                 BL 
                 cggtggtgcagatgaacttca 
                 1704 
               
               
                   
               
               
                 EGFP0012 
                 BL 
                 catggcggacttgaagaagtc 
                 1705 
               
               
                   
               
               
                 EGFP0013 
                 BL 
                 cgtcctccttgaagtcgatgc 
                 1706 
               
               
                   
               
            
           
         
       
     
     Table 2C shows the alpha-ENaC expression in cynomologous species for siRNA examples. 
     Example 3.4 Screening for Interferon-α induction. To evaluate the ability of siRNA to stimulate interferon-α (IFNα) release, siRNA was incubated with freshly purified peripheral blood mononuclear cells (PBMCs) in vitro for 24 hours. The siRNA was added either directly to PBMCs, or first complexed with a lipid transfection agent (GenePorter 2 or Lipofectamine 2000 or DOTAP transfection agent) and subsequently incubated with PBMCs. As positive controls for IFNα induction, unmodified control sequences DI_A_2216 and DI_A_5167 were included. 
     
       
         
           
               
            
               
                 DI_A_2216: is a single-stranded antisense 
               
               
                 DNA molecule 
               
               
                 (SEQ ID NO: 1707) 
               
               
                 5′-dGsdGsdGdGdGdAdCdGdAdTdCdGdTdCdGsdGsdGsdGsdGsd 
               
               
                   
               
               
                 G-3′ 
               
               
                   
               
               
                 DI_A_5167 is a cholesterol-conjugated siRNA 
               
               
                 5′-GUCAUCACACUGAAUACCAAU-s-chol-3′ 
               
               
                   
               
               
                 (SEQ ID NO: 1708) 
               
               
                 3′-Cs A s C AGUAGUGUGACUUAUGGUUA-5′ 
               
            
           
         
       
     
     After 24 hours, the IFNα was measured by ELISA. The basal IFNα level was determined for untreated cells and was always very close to a water-only control. The addition of transfection agent alone gave no or little increase of IFNα levels. Known stimulatory oligonucleotides were added to cells, either directly or in the presence of transfectant, and the expected increases of IFNα were observed. This setup allows to determine the stimulation of IFNα in human PBMC by siRNA (or other oligonucleotides). 
     Isolation of Human PBMCs: A concentrated fraction of leukocytes (buffy coat) was obtained from the Blood Bank Suhl, Institute for Transfusion Medicine, Germany. These cells were negative for a variety of pathogens, including HIV, HCV, and others. The buffy coat was diluted 1:1 with PBS, added to a tube containing Ficoll, and centrifuged for 20 minutes at 2200 rpm to allow fractionation. This was followed by removal of the turbid layer of white blood cells and transferred to a tube with fresh PBS and Ficoll, which was centrifuged for 15 minutes at 2200 rpm. The turbid layer of white blood cells was again removed, transferred to RPMI 1640 culture medium and centrifuged for 5 minutes at 1,200 rpm to pellet the white blood cells. The cells were resuspended in RPMI, pelleted as above, and resuspended in media with 10% FCS to 1×10 6 /mL. 
     Interferon-α Measurement: Cells in culture were combined with either 500 nM (or 1 μM) oligonucleotide in Optimem or 133 nM oligonucleotide in GP2 or Lipofectamine2000 or DOTAP transfection agent for 24 hours at 37° C. Interferon-α was measured with Bender Med Systems (Vienna, Austria) instant ELISA kit according to the manufacturer&#39;s instructions. 
     Selection of lead therapeutic sequences was based on a level of IFNα induction of less than 15% of the positive control. 
     Example 3.5 Determination of siRNA Stability in Sputum of CF Patients 
     Sputum samples were collected by Dr. Ahmet Uluer, Children&#39;s Hospital Boston. After collection, sputum samples were treated with antibiotic and were UV-irradiated to reduce bacterial content. To determine siRNA stability in sputum samples, siRNAs were incubated in 30 μL sputum at a concentration of 5 μM at 37° C. for the indicated times. The reaction was terminated by addition of proteinase K and samples were incubated at 42° C. for another 20 minutes. A 40-mer RNA molecule made of L-nucleotides (“Spiegelmer”) resistant to nuclease degradation was added and served as calibration standard. Samples were filtered through a 0.2 μm membrane to remove remaining debris. Samples were analyzed by denaturing ion exchange HPLC on a DNAPac PA 200 column (Dionex) at pH 11.0 using a gradient of sodium perchlorate for elution. siRNAs and degradation products were quantified by determination of the area under the peak for each sample. Concentration was normalized to the concentration in the un-incubated samples. 
     The selection of the lead therapeutic sequences (ND8356, ND8357 and ND8396) was based on an observed in vitro stability in CF sputum with a half-life greater than 60 minutes. 
     Example 3.6 Cross-Checking of Lead Therapeutic Sequences Against Known Polymorphisms in Human SCNN1A Gene 
     To exclude known polymorphisms from the target sites of lead candidates, lead siRNA sequences were checked against the NCBI single nucleotide polymorphism (SNP) database (http://www.ncbi.nlm.nih.gov/entrez/query.fcgi? CMD=search&amp;DB=snp). Of the 10 known exon polymorphisms in the human SCNN1A gene, none were shown to be present in the target sites of any of the 10 most potent lead therapeutic candidates. 
     Example 3.7: In Vitro Profiling of Top 5 Predicted Off-Target Sequences 
     A list of alignments for each sequence was sorted by homology over the 19-mer region. Off-targets were scored based on the number and position of the mismatches in accordance with the criteria described in example 1. The top 5 off-target sequences were identified for each lead therapeutic sequences (ND8356, ND8357 and ND8396). On- and off-target sequences were individually cloned into a dual luciferase reporter system. Each cloned fragment encompassed the target 19 nucleotides in addition to 10 nucleotides flanking region, both 5′ and 3′ of the target sequence. The fragments were cloned into a multiple cloning site 3′ to the  Renilla  luciferase sequence, under the control of an SV40 promoter. The activity of each siRNA against both on- and off-target sequences was determined by the relative fluorescence of the target  Renilla  luciferase to the Firefly luciferase, the latter being independently controlled by the HSV-TK promoter. Initially, transfections were performed in COS-7 cells at an siRNA concentration of 50 nM. Luciferase readouts were taken at 24 h post-transfection. At this high concentration of siRNA, no knockdown of greater than 30% was observed against any off-target sequence for any of the three lead siRNAs. Activity against the on-target sequence was demonstrated with a relative reduction in  Renilla  luciferase activity of approximately 80%. IC50 curves were also generated for each siRNA against the on-target sequence and controlled with the off-target sequences identified above. For each lead siRNA, on-target IC50&#39;s in this reporter assay were of similar order of magnitude (10-50 pM) to the IC50&#39;s obtained against the endogenous target in H441 (Example 3.3) indicating that for ND8356, ND8357 and ND8396, potency against the on-target sequence was at least 1000-5000 fold higher than for any of the predicted off-target sequences. 
     Example 3.8: Genotoxicity Profiling. Cytotoxicity Determination: Cytotoxicity was Determined by Using a Cell Counter for the Assessment of Culture Cell Number 
     It is well known that testing cytotoxic concentrations in vitro may induce genotoxic effects such as micronucleus formation. Therefore, we considered increased numbers of cells containing micronuclei appearing at cell counts of around 50% or less (compared to the concurrent negative control) to be cytotoxicity-related if no dose-dependent increase in micronucleated cells could already be observed at concentrations showing moderate toxicity at most. The analysis of a concentration showing at least 50% reduction in cell count is required by the guidelines regulating in vitro mammalian cell assays (OECD and ICH guidelines for the conduction of chromosome aberration testing). In addition, OECD protocols require testing of non-toxic compounds to include at least one precipitating concentration (as long as this doesn&#39;t exceed 10 mM or 5 mg/ml, whichever is lower). Since the in vitro micronucleus test aims to predict the outcome of the regulatory assays, i.e. in vitro chromosomal aberration test, the protocol for the in vitro micronucleus test was designed to meet the requirements for these tests. 
     Test system: TK6 cells are Ebstein-Barr-Virus transfected and immortalized cells (human lymphoblastoid origin derived from the spleen). Determination of the clastogenic and/or aneugenic potential in the micronucleus test in vitro with TK6 cells with/without S9-liver homogenate (2%) from male rats (Aroclor 1254-pretreated). Treatment time: 20 hr (−S9), 3 hr (+S9). Sampling time: 24 hr after the start of 3-hour treatment, 48 hr after the start of 20-hour treatment. For each substance at least three concentrations (2 cultures per concentration) and 2000 cells per concentration were analyzed. 
     The micronucleus inducing effect for a tested concentration was considered positive if the frequency of micronucleated cells was
         &gt;=2% and showed at least a doubling of the concurrent solvent control value, OR   &lt;2% and showed at least a 3-fold increase over the concurrent solvent control value       

     To conclude an experiment to be positive, dose-effect relationship and cytotoxicity have to be taken into account. 
     Summary: Lead therapeutic sequences ND8396, ND8356, ND8357 neither induced increased numbers of cells containing micronuclei after 20-hour treatment without metabolic activation, nor after 3-hour treatment with or without S9. No cytotoxic concentration could be analyzed up to the testing limit of 5 mg/ml. 
     Example 3.9 In Vitro Functional Efficacy in H441: ND8396 
     In order to demonstrate in vitro functional efficacy of lead siRNA against alphaENaC H441 cells were transfected with siRNA and prepared for Ussings chamber analysis of ion transport. For transfection, H441 cells were plated into T25 flasks at 2×10 6  cells per flask in culture medium supplemented with 200 nM Dexamethasone. Cells in each flask were transfected with either ND8396 or a non targeting control siRNA at 30 nM siRNA and 4 mL/mL Lipofectamine 2000 in a total volume of 5 mL (serum free medium). One day after transfection, cells were plated onto 1 cm 2  Snapwell inserts at confluency (2×10 5  cells per insert) to minimise the time required for differentiation and formation of tight junctions and supplied with medium in both the apical and the basolateral chambers. After one additional day of culture the apical medium was removed and the basolateral medium replaced, thus taking the cells to air-liquid interface (ALI) culture. Cells were maintained at ALI for a further six days prior to ion transport analysis. 
     For functional analysis in Ussings chambers, the Snapwell inserts were mounted in Vertical Diffusion Chambers (Costar) and were bathed with continuously gassed Ringer solution (5% CO 2  in 02; pH 7.4) maintained at 37° C. containing: 120 mM NaCl, 25 mM NaHCO 3 , 3.3 mM KH 2 PO 4 , 0.8 mM K 2 HPO 4 , 1.2 mM CaCl 2 , 1.2 mM MgCl 2 , and 10 mM glucose. The solution osmolarity was determined within the range of 280 and 300 mosmol/kgH 2 O. Cells were voltage clamped to 0 mV (model EVC4000; WPI). Transepithelial resistance (RT) was measured by applying a 1 or 2-mV pulse at 30-s intervals, or using the initial potential difference across the cells and the initial current measured, and then calculating RT by Ohm&#39;s law. Data were recorded using a PowerLab workstation (ADInstruments). Following siRNA treatment the basal characteristics of the cells and the amiloride-sensitive short circuit current (I SC  following application of 10 μM amiloride; apical side only) were recorded. ENaC channel activity in each culture was determined by the amiloride-sensitive I SC  in each case. 
     Following assay, cells on the individual inserts were lysed for RNA analysis. A knockdown of 75% at the RNA level at the time of assay (ND8396 as compared to non-targeting control) was correlated with a functional knockdown of the amiloride sensitive current of approximately 30% (ND8396 as compared to non-targeting control). 
     Nucleic acid sequences are represented below using standard nomenclature, and specifically the abbreviations of Table A. 
     
       
         
           
               
             
               
                 TABLE A 
               
             
            
               
                   
               
               
                 Abbreviations of nucleotide monomers used in nucleic 
               
               
                 acid sequence representation. It will be understood 
               
               
                 that these monomers, when present in an oligonucleotide, 
               
               
                 are mutually linked by 5′-3′-phosphodiester bonds. 
               
            
           
           
               
               
               
            
               
                   
                 Abbreviation a   
                 Nucleotide(s) 
               
               
                   
                   
               
               
                   
                 A 
                 adenosine-5′-phosphate 
               
               
                   
                 C 
                 cytidine-5′-phosphate 
               
               
                   
                 G 
                 guanosine-5′-phosphate 
               
               
                   
                 T 
                 2′-deoxy-thymidine-5′-phosphate 
               
               
                   
                 U 
                 uridine-5′-phosphate 
               
               
                   
                 c 
                 2′-O-methylcytidine-5′-phosphate 
               
               
                   
                 u 
                 2′-O-methyluridine-5′-phosphate 
               
               
                   
                 Ts 
                 2′-deoxy-thymidine-5′-phosphorothioate 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 1A 
               
             
            
               
                   
               
               
                 Selected siRNAs in initial screening set (human-rhesus ENaC alpha cross- 
               
               
                 reactive siRNAs). A total of 152 iRNA sequences were identified as an 
               
               
                 initial screening set, both with (sequence strands 1-304) and without 
               
               
                 (sequence strands 305-608) backbone modification. iRNA sequences were 
               
               
                 designed to be fully complementary to both the human and rhesus monkey 
               
               
                 alpha-ENaC sequences, according to the design criteria described in the 
               
               
                 examples section. The percentage residual expression of alpha-ENaC in 
               
               
                 two independent single-dose transfection experiments is shown (refer to 
               
               
                 examples section for methods used). 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 1st 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 screen 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 single 
                   
                 2nd 
                   
               
               
                   
                   
                   
                   
                   
                 dose @ 
                   
                 screen @ 
                   
               
               
                 Duplex 
                 Seq 
                   
                 Seq 
                   
                 60 nM in 
                   
                 60 nM 
                   
               
               
                 ID 
                 ID 
                 Sense 
                 ID 
                 Antisense 
                 H441; MV 
                 SD 
                 in H441 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 ND8285 
                 1 
                 AGcccGuAGcGuGGccuccTgT 
                 2 
                 GGAGGCcACGCuACGGGCUTgT 
                 92% 
                 4% 
                 114%  
                 16%  
               
               
                   
               
               
                 ND8286 
                 3 
                 ccGGcuAAuGGuGcAcGGGTgT 
                 4 
                 CCCGUGcACcAUuACCCGGTgT 
                 60% 
                 1% 
                 84% 
                 4% 
               
               
                   
               
               
                 ND8287 
                 5 
                 AuGcuAucGcGAcAGAAcATgT 
                 6 
                 UGUUCUGUCGCGAuAGcAUTgT 
                 27% 
                 2% 
                 35% 
                 3% 
               
               
                   
               
               
                 ND8288 
                 7 
                 uGcuAucGcGAcAGAAcAATgT 
                 8 
                 UUGUUCUGUCGCGAuAGcATgT 
                 23% 
                 1% 
                 32% 
                 4% 
               
               
                   
               
               
                 ND8289 
                 9 
                 GcccGuuuAuGuAuGcuccTgT 
                 10 
                 GGAGcAuAcAuAAACGGGCTgT 
                 64% 
                 8% 
                 93% 
                 8% 
               
               
                   
               
               
                 ND8290 
                 11 
                 GcccGuAGcGuGGccuccATgT 
                 12 
                 UGGAGGCcACGCuACGGGCTgT 
                 83% 
                 2% 
                 113%  
                 6% 
               
               
                   
               
               
                 ND8291 
                 13 
                 ccGGAAAuuAAAGAGGAGcTgT 
                 14 
                 GCUCCUCUUuAAUUUCCGGTgT 
                 54% 
                 2% 
                 79% 
                 10%  
               
               
                   
               
               
                 ND8292 
                 15 
                 ccGAAGGuuccGAAGccGATgT 
                 16 
                 UCGGCUUCGGAACCUUCGGTgT 
                 40% 
                 1% 
                 54% 
                 8% 
               
               
                   
               
               
                 ND8293 
                 17 
                 GcAAuucGGccuGcuuuucTgT 
                 18 
                 GAAAAGcAGGCCGAAUUGCTgT 
                 41% 
                 2% 
                 51% 
                 4% 
               
               
                   
               
               
                 ND8294 
                 19 
                 GGcGAAuuAcucucAcuucTgT 
                 20 
                 GAAGUGAGAGuAAUUCGCCTgT 
                 19% 
                 1% 
                 25% 
                 6% 
               
               
                   
               
               
                 ND8295 
                 21 
                 GcGAAuuAcucucAcuuccTgT 
                 22 
                 GGAAGUGAGAGuAAUUCGCTgT 
                 19% 
                 5% 
                 20% 
                 1% 
               
               
                   
               
               
                 ND8296 
                 23 
                 AAccAGGcGAAuuAcucucTgT 
                 24 
                 GAGAGuAAUUCGCCUGGUUTgT 
                 92% 
                 4% 
                 115%  
                 19%  
               
               
                   
               
               
                 ND8297 
                 25 
                 GGuAAuGGuGcAcGGGcAGTgT 
                 26 
                 CUGCCCGUGcACcAUuACCTgT 
                 70% 
                 2% 
                 108%  
                 14%  
               
               
                   
               
               
                 ND8298 
                 27 
                 cucAcGAuGGcccucGGuGTgT 
                 28 
                 cACCGAGGGCcAUCGUGAGTgT 
                 61% 
                 3% 
                 97% 
                 14%  
               
               
                   
               
               
                 ND8299 
                 29 
                 GcuccGAAGGuuccGAAGcTgT 
                 30 
                 GCUUCGGAACCUUCGGAGCTgT 
                 16% 
                 2% 
                 19% 
                 3% 
               
               
                   
               
               
                 ND8300 
                 31 
                 GccGAuAcuGGucuccAGGTgT 
                 32 
                 CCUGGAGACcAGuAUCGGCTgT 
                 60% 
                 5% 
                 65% 
                 5% 
               
               
                   
               
               
                 ND8301 
                 33 
                 ccGAuAcuGGucuucAGGcTgT 
                 34 
                 GCCUGGAGACcAGuAUCGGTgT 
                 63% 
                 2% 
                 65% 
                 6% 
               
               
                   
               
               
                 ND8302 
                 35 
                 uGcuGuuGcAccAuAcuuuTgT 
                 36 
                 AAAGuAUGGUGcAAcAGcATgT 
                 19% 
                 1% 
                 25% 
                 3% 
               
               
                   
               
               
                 ND8303 
                 37 
                 AAcGGucuGucccuGAuGcTgT 
                 38 
                 GcAUcAGGGAuAGACCGUUTgT 
                 90% 
                 3% 
                 96% 
                 11%  
               
               
                   
               
               
                 ND8304 
                 39 
                 uuAAcuuGcGGccuGGcGuTgT 
                 40 
                 ACGCcAGGCCGcAAGUuAATgT 
                 97% 
                 3% 
                 101%  
                 11%  
               
               
                   
               
               
                 ND8305 
                 41 
                 GcuGGuuAcucAcGAuGGcTgT 
                 42 
                 GCcAUCGUGAGuAACcAGCTgT 
                 73% 
                 2% 
                 78% 
                 7% 
               
               
                   
               
               
                 ND8306 
                 43 
                 uuAcucAcGAuGGcccucGTgT 
                 44 
                 CGAGGGCcAUCGUGAGuAATgT 
                 91% 
                 7% 
                 93% 
                 6% 
               
               
                   
               
               
                 ND8307 
                 45 
                 GAAGccGAuAcuGGucuccTgT 
                 46 
                 GGAGACcAGuAUCGGCUUCTgT 
                 71% 
                 3% 
                 73% 
                 6% 
               
               
                   
               
               
                 ND8308 
                 47 
                 GAuAcuGGucuccAGGccGTgT 
                 48 
                 CGGCCUGGAGACcAGuAUCTgT 
                 86% 
                 1% 
                 90% 
                 9% 
               
               
                   
               
               
                 ND8309 
                 49 
                 AuAcuGGucuccAGGccGATgT 
                 50 
                 UCGGCCUGGAGACcAGuAUTgT 
                 71% 
                 5% 
                 70% 
                 8% 
               
               
                   
               
               
                 ND8310 
                 51 
                 cAAcGGucuGucccuGAuGTgT 
                 52 
                 cAUcAGGGAcAGACCGUUGTgT 
                 80% 
                 2% 
                 84% 
                 9% 
               
               
                   
               
               
                 ND8311 
                 53 
                 uuuAAcuuGcGGccuGGcGTgT 
                 54 
                 CGCcAGGCCGcAAGUuAAATgT 
                 95% 
                 2% 
                 107%  
                 15%  
               
               
                   
               
               
                 ND8312 
                 55 
                 uAcucAcGAuGGcccucGGTgT 
                 56 
                 CCGAGGGCcAUCGUGAGuATgT 
                 44% 
                 2% 
                 97% 
                 9% 
               
               
                   
               
               
                 ND8313 
                 57 
                 uuucGGAGAGuAcuucAGcTgT 
                 58 
                 GCUGAAGuACUCUCCGAAATgT 
                 14% 
                 2% 
                 16% 
                 2% 
               
               
                   
               
               
                 ND8314 
                 59 
                 GcAGAcGcucuuuGAccuGTgT 
                 60 
                 cAGGUcAAAGAGCGUCUGCTgT 
                 55% 
                 4% 
                 58% 
                 5% 
               
               
                   
               
               
                 ND8315 
                 61 
                 cuAcAucuucuAuccGcGGTgT 
                 62 
                 CCGCGGAuAGAAGAUGuAGTgT 
                 20% 
                 3% 
                 26% 
                 4% 
               
               
                   
               
               
                 ND8316 
                 63 
                 AGGcGAAuuAcucucAcuuTgT 
                 64 
                 AAGUGAGAGuAAUUCGCCUTgT 
                 24% 
                 1% 
                 25% 
                 2% 
               
               
                   
               
               
                 ND8317 
                 65 
                 ccGcuucAAccAGGucuccTgT 
                 66 
                 GGAGACCUGGUUGAAGCGGTgT 
                 62% 
                 5% 
                 64% 
                 4% 
               
               
                   
               
               
                 ND8318 
                 67 
                 cAAccGcAuGAAGAcGGccTgT 
                 68 
                 GGCCGUCUUcAUGCGGUUGTgT 
                 54% 
                 6% 
                 54% 
                 4% 
               
               
                   
               
               
                 ND8319 
                 69 
                 AuGAAGAcGGccuucuGGGTgT 
                 70 
                 CCcAGAAGGCCGUCUUcAUTgT 
                 44% 
                 4% 
                 44% 
                 6% 
               
               
                   
               
               
                 ND8320 
                 71 
                 AGcAcAAccGcAuGAAGAcTgT 
                 72 
                 GUCUUcAUGCGGUUGUGCUTgT 
                 15% 
                 1% 
                 16% 
                 1% 
               
               
                   
               
               
                 ND8321 
                 73 
                 ucGAGuuccAccGcuccuATgT 
                 74 
                 uAGGAGCGGUGGAACUCGATgT 
                 85% 
                 5% 
                 89% 
                 13%  
               
               
                   
               
               
                 ND8322 
                 75 
                 cuGcuucuAccAGAcAuAcTgT 
                 76 
                 GuAUGUCUGGuAGAAGcAGTgT 
                 46% 
                 4% 
                 44% 
                 3% 
               
               
                   
               
               
                 ND8323 
                 77 
                 GAGGGAGuGGuAccGcuucTgT 
                 78 
                 GAAGCGGuACcACUCCCUCTgT 
                 60% 
                 7% 
                 56% 
                 3% 
               
               
                   
               
               
                 ND8324 
                 79 
                 ccuuuAuGGAuGAuGGuGGTgT 
                 80 
                 CcACcAUcAUCcAuAAAGGTgT 
                 83% 
                 9% 
                 82% 
                 1% 
               
               
                   
               
               
                 ND8325 
                 81 
                 uGAGGGAGuGGuAccGcuuTgT 
                 82 
                 AAGCGGuACcACUCCCUcATgT 
                 77% 
                 6% 
                 72% 
                 2% 
               
               
                   
               
               
                 ND8326 
                 83 
                 ccuGcAAccAGGcGAAuuATgT 
                 84 
                 uAAUUCGCCUGGUUGcAGGTgT 
                 41% 
                 4% 
                 44% 
                 7% 
               
               
                   
               
               
                 ND8327 
                 85 
                 GGccuGGcGuGGAGAccucTgT 
                 86 
                 GAGGUCUCcACGCcAGGCCTgT 
                 101%  
                 5% 
                 95% 
                 4% 
               
               
                   
               
               
                 ND8328 
                 87 
                 uGcuuuucGGAGAGuAcuuTgT 
                 88 
                 AAGuACUCUCCGAAAAGcATgT 
                 96% 
                 1% 
                 39% 
                 3% 
               
               
                   
               
               
                 ND8329 
                 89 
                 cccGuAGcGuGGccuccAGTgT 
                 90 
                 CUGGAGGCcACGCuACGGGTgT 
                 50% 
                 1% 
                 51% 
                 2% 
               
               
                   
               
               
                 ND8330 
                 91 
                 ccGuAGcGuGGccuccAGcTgT 
                 92 
                 GCUGGAGGCcACGCuACGGTgT 
                 86% 
                 9% 
                 84% 
                 3% 
               
               
                   
               
               
                 ND8331 
                 93 
                 ccAGGcGAAuuAcucucAcTgT 
                 94 
                 GUGAGAGuAAUUCGCCUGGTgT 
                 16% 
                 2% 
                 13% 
                 1% 
               
               
                   
               
               
                 ND8332 
                 95 
                 GAAAcuGcuAuAcuuucAATgT 
                 96 
                 UUGAAAGuAuAGcAGUUUCTgT 
                 10% 
                 1% 
                 10% 
                 1% 
               
               
                   
               
               
                 ND8333 
                 97 
                 GcccGGGuAAuGGuGcAcGTgT 
                 98 
                 CGUGcACcAUuACCCGGGCTgT 
                 83% 
                 6% 
                 82% 
                 4% 
               
               
                   
               
               
                 ND8334 
                 99 
                 cccGGGuAAuGGuGcAcGGTgT 
                 100 
                 CCGUGcACcAUuACCCGGGTgT 
                 56% 
                 4% 
                 71% 
                 10%  
               
               
                   
               
               
                 ND8335 
                 101 
                 cGGGuAAuGGuGcAcGGGcTgT 
                 102 
                 GCCCGUGcACcAUuACCCGTgT 
                 42% 
                 3% 
                 91% 
                 8% 
               
               
                   
               
               
                 ND8336 
                 103 
                 GGGuAAuGGuGcAcGGGcATgT 
                 104 
                 UGCCCGUGcACcAUuACCCTgT 
                 65% 
                 5% 
                 71% 
                 7% 
               
               
                   
               
               
                 ND8337 
                 105 
                 uAAuGGuGcAcGGGcAGGATgT 
                 106 
                 UCCUGCCCGUGcACcAUuATgT 
                 46% 
                 3% 
                 46% 
                 4% 
               
               
                   
               
               
                 ND8338 
                 107 
                 cuGGuuAcucAcGAuGGccTgT 
                 108 
                 GGCcAUCGUGAGuAACcAGTgT 
                 74% 
                 5% 
                 79% 
                 10%  
               
               
                   
               
               
                 ND8339 
                 109 
                 GuuAcucAcGAuGGcccucTgT 
                 110 
                 GAGGGCcAUCGUGAGuAACTgT 
                 85% 
                 6% 
                 92% 
                 8% 
               
               
                   
               
               
                 ND8340 
                 111 
                 uGucAcGAuGGucAcccucTgT 
                 112 
                 GAGGGUGACcAUCGUGAcATgT 
                 85% 
                 4% 
                 74% 
                 5% 
               
               
                   
               
               
                 ND8341 
                 113 
                 uGcuccGAAGGuuccGAAGTgT 
                 114 
                 CUUCGGAACCUUCGGAGcATgT 
                 37% 
                 2% 
                 32% 
                 3% 
               
               
                   
               
               
                 ND8342 
                 115 
                 uccGAAGGuuccGAAGccGTgT 
                 116 
                 CGGCUUCGGAACCUUCGGATgT 
                 60% 
                 4% 
                 47% 
                 5% 
               
               
                   
               
               
                 ND8343 
                 117 
                 uuccGAAGccGAuAcuGGuTgT 
                 118 
                 ACcAGuAUCGGCUUCGGAATgT 
                 15% 
                 1% 
                 13% 
                 2% 
               
               
                   
               
               
                 ND8344 
                 119 
                 AGccGAuAcuGGucuccAGTgT 
                 120 
                 CUGGAGACcAGuAUCGGCUTgT 
                 49% 
                 3% 
                 41% 
                 3% 
               
               
                   
               
               
                 ND8345 
                 121 
                 cuuGGuAcuGccucuGAAcTgT 
                 122 
                 GUUcAGAGGcAGuACcAAGTgT 
                 55% 
                 2% 
                 47% 
                 4% 
               
               
                   
               
               
                 ND8346 
                 123 
                 cucccGuAGcAcAcuAuAATgT 
                 124 
                 UuAuAGUGUGCuACGGGAGTgT 
                 67% 
                 3% 
                 57% 
                 5% 
               
               
                   
               
               
                 ND8347 
                 125 
                 ucccGuAGcAcAcuAuAAcTgT 
                 126 
                 GUuAgAGUGUGCuACGGGATgT 
                 29% 
                 1% 
                 26% 
                 3% 
               
               
                   
               
               
                 ND8348 
                 127 
                 uGcAccAuAcuuucuuGuATgT 
                 128 
                 uAcAAGAAAGuAUGGUGcATgT 
                 17% 
                 1% 
                 15% 
                 3% 
               
               
                   
               
               
                 ND8349 
                 129 
                 uuGcccGuuuAuGuAuGcuTgT 
                 130 
                 AGcAgAcAuAAACGGGcAATgT 
                 68% 
                 2% 
                 50% 
                 4% 
               
               
                   
               
               
                 ND8350 
                 131 
                 uGcccGuuuAuGuAuGcucTgT 
                 132 
                 GAGcAuAcAuAAACGGGcATgT 
                 59% 
                 8% 
                 44% 
                 6% 
               
               
                   
               
               
                 ND8351 
                 133 
                 GGAcccuAGAccucuGcAGTgT 
                 134 
                 CUGcAGAGGUCuAGGGUCCTgT 
                 86% 
                 11%  
                 82% 
                 2% 
               
               
                   
               
               
                 ND8352 
                 135 
                 ccuAGAccucuGcAGcccATgT 
                 136 
                 UGGGCUGcAGAGGUCuAGGTgT 
                 69% 
                 7% 
                 79% 
                 3% 
               
               
                   
               
               
                 ND8353 
                 137 
                 uGGcAuGAuGuAcuGGcAATgT 
                 138 
                 UUGCcAGuAcAUcAUGCcATgT 
                 58% 
                 4% 
                 52% 
                 4% 
               
               
                   
               
               
                 ND8354 
                 139 
                 uAcuGGcAAuucGGccuGcTgT 
                 140 
                 GcAGGCCGAAUUGCcAGuATgT 
                 101%  
                 4% 
                 100%  
                 4% 
               
               
                   
               
               
                 ND8355 
                 141 
                 AAuucGGccuGcuuuucGGTgT 
                 142 
                 CCGAAAAGcAGGCCGAAUUTgT 
                 49% 
                 1% 
                 43% 
                 6% 
               
               
                   
               
               
                 ND8356 
                 143 
                 cuGcuuuucGGAGAGuAcuTgT 
                 144 
                 AGuACUCUCCGAAAAGcAGTgT 
                 17% 
                 3% 
                 18% 
                 1% 
               
               
                   
               
               
                 ND8357 
                 145 
                 uucGGAGAGuAcuucAGcuTgT 
                 146 
                 AGCUGAAGuACUCUCCGAATgT 
                 13% 
                 3% 
                 16% 
                 2% 
               
               
                   
               
               
                 ND8358 
                 147 
                 AGcAGAcGcucuuuGAccuTgT 
                 148 
                 AGGUcAAAGAGCGUCUGCUTgT 
                 73% 
                 9% 
                 71% 
                 5% 
               
               
                   
               
               
                 ND8359 
                 149 
                 cuuGcAGcGccuGAGuGucTgT 
                 150 
                 GACCCUcAGGCGCUGcAAGTgT 
                 57% 
                 9% 
                 64% 
                 7% 
               
               
                   
               
               
                 ND8360 
                 151 
                 uGGcuuuAAcuuGcGGccuTgT 
                 152 
                 AGGCCGcAAGUuAAAGCcATgT 
                 102%  
                 12%  
                 106%  
                 10%  
               
               
                   
               
               
                 ND8361 
                 153 
                 GcuuuAAcuuGcGGccuGGTgT 
                 154 
                 CcAGGCCGcAAGUuAAAGCTgT 
                 83% 
                 5% 
                 82% 
                 8% 
               
               
                   
               
               
                 ND8362 
                 155 
                 uAAcuuGcGGccuGGcGuGTgT 
                 156 
                 cACGCcAGGCCGcAAGUuATgT 
                 119%  
                 2% 
                 115%  
                 6% 
               
               
                   
               
               
                 ND8363 
                 157 
                 AccuuuAcccuucAAAGuATgT 
                 158 
                 uACUUUGAAGGGuAAAGGUTgT 
                 17% 
                 3% 
                 13% 
                 2% 
               
               
                   
               
               
                 ND8364 
                 159 
                 GGuuAcucAcGAuGGcccuTgT 
                 160 
                 AGGGCcAUCGUGAGuAACCTgT 
                 104%  
                 9% 
                 117%  
                 17%  
               
               
                   
               
               
                 ND8365 
                 161 
                 cAcGAuGGcccucGGuGAcTgT 
                 162 
                 GUcACCGAGGGCcAUCGUGTgT 
                 148%  
                 13%  
                 100%  
                 9% 
               
               
                   
               
               
                 ND8366 
                 163 
                 AGAuGcuAucGcGAcAGAATgT 
                 164 
                 UUCUGUCGCGAuAGcAUCUTgT 
                 46% 
                 2% 
                 70% 
                 6% 
               
               
                   
               
               
                 ND8367 
                 165 
                 AcGAuGGucAcccuccuGuTgT 
                 166 
                 AcAGGAGGGUGACcAUCGUTgT 
                 85% 
                 6% 
                 128%  
                 10%  
               
               
                   
               
               
                 ND8368 
                 167 
                 cuccGAAGGuuccGAAGccTgT 
                 168 
                 GGCUUCGGAACCUUCGGAGTgT 
                 12% 
                 2% 
                 18% 
                 1% 
               
               
                   
               
               
                 ND8369 
                 169 
                 AAGGuuccGAAGccGAuAcTgT 
                 170 
                 GuAUCGGCUUCGGAACCUUTgT 
                 63% 
                 7% 
                 114%  
                 19%  
               
               
                   
               
               
                 ND8370 
                 171 
                 GGuAcuGccucuGAAcAcuTgT 
                 172 
                 AGUGUUcAGAGGcAGuACCTgT 
                 36% 
                 3% 
                 71% 
                 6% 
               
               
                   
               
               
                 ND8371 
                 173 
                 AGcuuuGAcAAGGAAcuuuTgT 
                 174 
                 AAAGUUCCUUGUcAAAGCUTgT 
                 17% 
                 1% 
                 21% 
                 1% 
               
               
                   
               
               
                 ND8372 
                 175 
                 uuuGAcAAGGAAcuuuccuTgT 
                 176 
                 AGGAAAGUUCCUUGUcAAATgT 
                 16% 
                 2% 
                 26% 
                 4% 
               
               
                   
               
               
                 ND8373 
                 177 
                 uGAcAAGGAAcuuuccuAATgT 
                 178 
                 UuAGGAAAGUUCCUUGUcATgT 
                 12% 
                 1% 
                 22% 
                 5% 
               
               
                   
               
               
                 ND8374 
                 179 
                 cccGuAGcAcAcuAuAAcATgT 
                 180 
                 UGUuAuAGUGUGCuACGGGTgT 
                 41% 
                 2% 
                 75% 
                 3% 
               
               
                   
               
               
                 ND8375 
                 181 
                 cAcuAuAAcAucuGcuGGATgT 
                 182 
                 UCcAGcAGAUGUuAuAGUGTgT 
                 17% 
                 1% 
                 26% 
                 2% 
               
               
                   
               
               
                 ND8376 
                 183 
                 uuGcuGuuGcAccAuAcuuTgT 
                 184 
                 AAGuAUGGUGcAAcAGcAATgT 
                 45% 
                 4% 
                 69% 
                 6% 
               
               
                   
               
               
                 ND8377 
                 185 
                 GuAcuGGcAAuucGGccuGTgT 
                 186 
                 cAGGCCGAAUUGCcAGuACTgT 
                 60% 
                 6% 
                 120%  
                 8% 
               
               
                   
               
               
                 ND8378 
                 187 
                 uucGGccuGcuuuucGGAGTgT 
                 188 
                 CUCCGAAAAGcAGGCCGAATgT 
                 57% 
                 5% 
                 86% 
                 11%  
               
               
                   
               
               
                 ND8379 
                 189 
                 ccuGcuuuucGGAGAGuAcTgT 
                 190 
                 GuACUCUCCGAAAAGcAGGTgT 
                 43% 
                 5% 
                 50% 
                 3% 
               
               
                   
               
               
                 ND8380 
                 191 
                 GcuuuucGGAGAGuAcuucTgT 
                 192 
                 GAAGuACUCUCCGAAAAGCTgT 
                 16% 
                 2% 
                 24% 
                 2% 
               
               
                   
               
               
                 ND8381 
                 193 
                 cuuuucGGAGAGuAcuucATgT 
                 194 
                 UGAAGuACUCUCCGAAAAGTgT 
                 12% 
                 1% 
                 16% 
                 3% 
               
               
                   
               
               
                 ND8382 
                 195 
                 cAAccucAAcucGGAcAAGTgT 
                 196 
                 CUUGUCCGAGUUGAGGUUGTgT 
                 33% 
                 2% 
                 39% 
                 3% 
               
               
                   
               
               
                 ND8383 
                 197 
                 cuAccAGAcAuAcucAucATgT 
                 198 
                 UGAUGAGuAUGUCUGGuAGTgT 
                 13% 
                 1% 
                 23% 
                 6% 
               
               
                   
               
               
                 ND8384 
                 199 
                 cuGucGAGGcuGccAGAGATgT 
                 200 
                 UCUCUGGcAGCCUCGAcAGTgT 
                 11% 
                 1% 
                 18% 
                 3% 
               
               
                   
               
               
                 ND8385 
                 201 
                 AAAcuGcuAuAcuuucAAuTgT 
                 202 
                 AUUGAAAGuAuAGcAGUUUTgT 
                 48% 
                 6% 
                 64% 
                 11%  
               
               
                   
               
               
                 ND8386 
                 203 
                 GGcuuuAAcuuGcGGccuGTgT 
                 204 
                 cAGGCCGcAAGUuAAAGCCTgT 
                 55% 
                 7% 
                 70% 
                 8% 
               
               
                   
               
               
                 ND8387 
                 205 
                 cuuuAAcuuGcGGccuGGcTgT 
                 206 
                 GCcAGGCCGcAAGUuAAAGTgT 
                 40% 
                 11%  
                 87% 
                 14%  
               
               
                   
               
               
                 ND8388 
                 207 
                 AGGuGuGuAuucAcuccuGTgT 
                 208 
                 cAGGAGUGAAuAcAcACCUTgT 
                 45% 
                 3% 
                 41% 
                 5% 
               
               
                   
               
               
                 ND8389 
                 209 
                 AcGAuGGcccucGGuGAcATgT 
                 210 
                 UGUcACCGAGGGCcAUCGUTgT 
                 43% 
                 2% 
                 60% 
                 9% 
               
               
                   
               
               
                 ND8390 
                 211 
                 cuGAAcAcucuGGuuucccTgT 
                 212 
                 GGGAAACcAGAGUGUUcAGTgT 
                 33% 
                 2% 
                 48% 
                 11%  
               
               
                   
               
               
                 ND8391 
                 213 
                 cuAuAAcAucuGcuGGAGuTgT 
                 214 
                 ACUCcAGcAGAUGUuAuAGTgT 
                 16% 
                 1% 
                 17% 
                 4% 
               
               
                   
               
               
                 ND8392 
                 215 
                 GcAccAuAcuuucuuGuAcTgT 
                 216 
                 GuAcAAGAAAGuAUGGUGCTgT 
                 19% 
                 1% 
                 22% 
                 4% 
               
               
                   
               
               
                 ND8393 
                 217 
                 uGucuAGcccAucAuccuGTgT 
                 218 
                 cAGGAUGAUGGGCuAGAcATgT 
                 69% 
                 3% 
                 92% 
                 15%  
               
               
                   
               
               
                 ND8394 
                 219 
                 AGGAcccuAGAccucuGcATgT 
                 220 
                 UGcAGAGGUCuAGGGUCCUTgT 
                 94% 
                 5% 
                 86% 
                 13%  
               
               
                   
               
               
                 ND8395 
                 221 
                 ccAccGcuccuAccGAGAGTgT 
                 222 
                 CUCUCGGuAGGAGCGGUGGTgT 
                 55% 
                 1% 
                 65% 
                 6% 
               
               
                   
               
               
                 ND8396 
                 223 
                 uAccGAGAGcucuucGAGuTgT 
                 224 
                 ACUCGAAGAGCUCUCGGuATgT 
                 11% 
                 1% 
                 11% 
                 1% 
               
               
                   
               
               
                 ND8397 
                 225 
                 AAcAuccuGucGAGGcuGcTgT 
                 226 
                 GcAGCCUCGAcAGGAUGUUTgT 
                 90% 
                 7% 
                 72% 
                 11%  
               
               
                   
               
               
                 ND8398 
                 227 
                 GAAccuuuAcccuucAAAGTgT 
                 228 
                 CUUUGAAGGGuAAAGGUUCTgT 
                 22% 
                 2% 
                 25% 
                 4% 
               
               
                   
               
               
                 ND8399 
                 229 
                 GGuuccGAAGccGAuAcuGTgT 
                 230 
                 cAGuAUCGGCUUCGGAACCTgT 
                 93% 
                 9% 
                 89% 
                 9% 
               
               
                   
               
               
                 ND8400 
                 231 
                 AAGccGAuAcuGGucuccATgT 
                 232 
                 UGGAGACcAGuAUCGGCUUTgT 
                 35% 
                 2% 
                 42% 
                 9% 
               
               
                   
               
               
                 ND8401 
                 233 
                 ucuAGcccAucAuccuGcuTgT 
                 234 
                 AGcAGGAUGAUGGGCuAGATgT 
                 95% 
                 6% 
                 95% 
                 14%  
               
               
                   
               
               
                 ND8402 
                 235 
                 cGGcGccAuccGccuGGuGTgT 
                 236 
                 cACcAGGCGGAUGGCGCCGTgT 
                 81% 
                 8% 
                 89% 
                 17%  
               
               
                   
               
               
                 ND8403 
                 237 
                 uuuucGGAGAGuAcuucAGTgT 
                 238 
                 CUGAAGuACUCUCCGAAAATgT 
                 13% 
                 1% 
                 13% 
                 1% 
               
               
                   
               
               
                 ND8404 
                 239 
                 GAGAGuAcuucAGcuAcccTgT 
                 240 
                 GGGuAGCUGAAGuACUCUCTgT 
                 71% 
                 3% 
                 100%  
                 10%  
               
               
                   
               
               
                 ND8405 
                 241 
                 GAcGcucuuuGAccuGuAcTgT 
                 242 
                 GuAcAGGUcAAAGAGCGUCTgT 
                 84% 
                 5% 
                 92% 
                 13%  
               
               
                   
               
               
                 ND8406 
                 243 
                 uGuGuAuucAcuccuGcuuTgT 
                 244 
                 AAGcAGGAGUGAAuAcAcATgT 
                 78% 
                 2% 
                 89% 
                 8% 
               
               
                   
               
               
                 ND8407 
                 245 
                 AAcAAcAAGAGAAAuGGAGTgT 
                 246 
                 CUCcAUUUCUCUUGUUGUUTgT 
                 66% 
                 3% 
                 66% 
                 21%  
               
               
                   
               
               
                 ND8408 
                 247 
                 AuuGAAGGAuGuGcAGGGcTgT 
                 248 
                 GCCCUGcAcAUCCUUcAAUTgT 
                 25% 
                 1% 
                 36% 
                 6% 
               
               
                   
               
               
                 ND8409 
                 249 
                 ucucAGAGccGcccAAAcuTgT 
                 250 
                 AGUUUGGGCGGCUCUGAGATgT 
                 18% 
                 1% 
                 24% 
                 2% 
               
               
                   
               
               
                 ND8410 
                 251 
                 AAAcAcAAccAAGGGuAcATgT 
                 252 
                 UGuACCCUUGGUUGUGUUUTgT 
                 21% 
                 1% 
                 35% 
                 2% 
               
               
                   
               
               
                 ND8411 
                 253 
                 uAcccGuGcccucAcAGAGTgT 
                 254 
                 CUCUGUGAGGGcACGGGuATgT 
                 57% 
                 2% 
                 67% 
                 4% 
               
               
                   
               
               
                 ND8412 
                 255 
                 uAGcAcAcuAuAAcAucuGTgT 
                 256 
                 cAGAUGUuAuAGUGUGCuATgT 
                 30% 
                 2% 
                 41% 
                 1% 
               
               
                   
               
               
                 ND8413 
                 257 
                 GGuGuGuAuucAcuccuGcTgT 
                 258 
                 GcAGGAGUGAAuAcAcACCTgT 
                 73% 
                 1% 
                 90% 
                 9% 
               
               
                   
               
               
                 ND8414 
                 259 
                 cAuGAucAAGGAGuGuGGcTgT 
                 260 
                 GCcAcACUCCUUGAUcAUGTgT 
                 65% 
                 2% 
                 67% 
                 5% 
               
               
                   
               
               
                 ND8415 
                 261 
                 AcucAcGAuGGcccucGGuTgT 
                 262 
                 ACCGAGGGCcAUCGUGAGUTgT 
                 96% 
                 6% 
                 95% 
                 6% 
               
               
                   
               
               
                 ND8416 
                 263 
                 GGAGcuuuGAcAAGGAAcuTgT 
                 264 
                 AGUUCCUUGUcAAAGCUCCTgT 
                 24% 
                 1% 
                 28% 
                 4% 
               
               
                   
               
               
                 ND8417 
                 265 
                 AuAcccGuGcccucAcAGATgT 
                 266 
                 UCUGUGAGGGcACGGGuAUTgT 
                 54% 
                 1% 
                 62% 
                 2% 
               
               
                   
               
               
                 ND8418 
                 267 
                 GGAGuGGccAAAGucAAcATgT 
                 268 
                 UGUUGACUUUGGCcACUCCTgT 
                 93% 
                 2% 
                 86% 
                 11%  
               
               
                   
               
               
                 ND8419 
                 269 
                 AAcuAcAAAAccAAuucuGTgT 
                 270 
                 cAGAAUUGGUUUUGuAGUUTgT 
                 101%  
                 5% 
                 108%  
                 19%  
               
               
                   
               
               
                 ND8420 
                 271 
                 uGcuGGAGuGuuGcuGuuGTgT 
                 272 
                 cAAcAGcAAcACUCcAGcATgT 
                 29% 
                 1% 
                 26% 
                 1% 
               
               
                   
               
               
                 ND8421 
                 273 
                 AGGucuccuGcAAccAGGcTgT 
                 274 
                 GCCUGGUUGcAGGAGACCUTgT 
                 95% 
                 10%  
                 91% 
                 17%  
               
               
                   
               
               
                 ND8422 
                 275 
                 cuuuGGcAuGAuGuAcuGGTgT 
                 276 
                 CcAGuAcAUcAUGCcAAAGTgT 
                 86% 
                 3% 
                 84% 
                 6% 
               
               
                   
               
               
                 ND8423 
                 277 
                 cAucuGcAcccucAAucccTgT 
                 278 
                 GGGAUUGAGGGUGcAGAUGTgT 
                 82% 
                 11%  
                 73% 
                 4% 
               
               
                   
               
               
                 ND8424 
                 279 
                 cGAcuGcAccAAGAAuGGcTgT 
                 280 
                 GCcAUUCUUGGUGcAGUCGTgT 
                 70% 
                 8% 
                 69% 
                 7% 
               
               
                   
               
               
                 ND8425 
                 281 
                 AAAAcAcAAccAAGGGuAcTgT 
                 282 
                 GuACCCUUGGUUGUGUUUUTgT 
                 95% 
                 6% 
                 106%  
                 12%  
               
               
                   
               
               
                 ND8426 
                 283 
                 cAucuGcuGGAGuGuuGcuTgT 
                 284 
                 AGcAAcACUCcAGcAGAUGTgT 
                 30% 
                 2% 
                 37% 
                 1% 
               
               
                   
               
               
                 ND8427 
                 285 
                 ccuAcAucuucuAuccGcGTgT 
                 286 
                 CGCGGAuAGAAGAUGuAGGTgT 
                 42% 
                 6% 
                 30% 
                 1% 
               
               
                   
               
               
                 ND8428 
                 287 
                 GccuAcAucuucuAuccGcTgT 
                 288 
                 GCGGAuAGAAGAUGuAGGCTgT 
                 65% 
                 7% 
                 54% 
                 3% 
               
               
                   
               
               
                 ND8429 
                 289 
                 GAGuGGuAccGcuuccAcuTgT 
                 290 
                 AGUGGAAGCGGuACcACUCTgT 
                 95% 
                 11%  
                 86% 
                 19%  
               
               
                   
               
               
                 ND8430 
                 291 
                 GGuAccGcuuccAcuAcAuTgT 
                 292 
                 AUGuAGUGGAAGCGGuACCTgT 
                 111%  
                 19%  
                 96% 
                 14%  
               
               
                   
               
               
                 ND8431 
                 293 
                 GuGGuAccGcuuccAcuAcTgT 
                 294 
                 GuAGUGGAAGCGGuACcACTgT 
                 98% 
                 13%  
                 52% 
                 26%  
               
               
                   
               
               
                 ND8432 
                 295 
                 GAAuuAcucucAcuuccAcTgT 
                 296 
                 GGUGGAAGUGAGAGuAAUUTgT 
                 111%  
                 21%  
                 73% 
                 27%  
               
               
                   
               
               
                 ND8433 
                 297 
                 AAuuAcucucAcuuccAccTgT 
                 298 
                 GGUGGAAGUGAGAGuAAUUTgT 
                 109%  
                 22%  
                 105%  
                 7% 
               
               
                   
               
               
                 ND8434 
                 299 
                 uAcucucAcuuccAccAccTgT 
                 300 
                 GGUGGUGGAAGUGAGAGuATgT 
                 106%  
                 23%  
                 95% 
                 7% 
               
               
                   
               
               
                 ND8435 
                 301 
                 AGuGGuAccGcuuccAcuATgT 
                 302 
                 uAGUGGAAGCGGuACcACUTgT 
                 109%  
                 18%  
                 102%  
                 9% 
               
               
                   
               
               
                 ND8436 
                 303 
                 GGGcAAcuucAucuucGccTgT 
                 304 
                 GGCGAAGAUGAAGUUGCCCTgT 
                 109%  
                 18%  
                 107%  
                 14%  
               
               
                   
               
               
                 ND-8501 
                 305 
                 AGCCCGUAGCGUGGCCUCCTgT 
                 306 
                 GGAGGCCACGCUACGGGCUTgT 
                 84% 
                 14%  
                 69% 
                 3% 
               
               
                   
               
               
                 ND-8502 
                 307 
                 CCGGGUAAUGGUGCACGGGTgT 
                 308 
                 CCCGUGCACCAUUACCCGGTgT 
                 41% 
                 6% 
                 30% 
                 2% 
               
               
                   
               
               
                 ND-8503 
                 309 
                 AUGCUAUCGCGACAGAACATgT 
                 310 
                 UGUUCUGUCGCGAUAGCAUTgT 
                 11% 
                 2% 
                 10% 
                 2% 
               
               
                   
               
               
                 ND-8504 
                 311 
                 UGCUAUCGCGACAGAACAATgT 
                 312 
                 UUGUUCUGUCGCGAUAGCATgT 
                 19% 
                 2% 
                 10% 
                 0% 
               
               
                   
               
               
                 ND-8505 
                 313 
                 GCCCGUUUAUGUAUGCUCCTgT 
                 314 
                 GGAGCAUACAUAAACGGGCTgT 
                 23% 
                 3% 
                 16% 
                 1% 
               
               
                   
               
               
                 ND-8506 
                 315 
                 GCCCGUAGCGUGGCCUCCATgT 
                 316 
                 UGGAGGCCACGCUACGGGCTgT 
                 32% 
                 3% 
                 22% 
                 1% 
               
               
                   
               
               
                 ND-8507 
                 317 
                 CCGGAAAUUAAAGAGGAGCTgT 
                 318 
                 GCUCCUCUUUAAUUUCCGGTgT 
                 35% 
                 4% 
                 24% 
                 1% 
               
               
                   
               
               
                 ND-8508 
                 319 
                 CCGAAGGUUCCGAAGCCGATgT 
                 320 
                 UCGGCUUCGGAACCUUCGGTgT 
                 19% 
                 2% 
                 13% 
                 1% 
               
               
                   
               
               
                 ND-8509 
                 321 
                 GCAAUUCGGCCUGCUUUUCTgT 
                 322 
                 GAAAAGCAGGCCGAAUUGCTgT 
                 12% 
                 1% 
                  8% 
                 1% 
               
               
                   
               
               
                 ND-8510 
                 323 
                 GGCGAAUUACUCUCACUUCTgT 
                 324 
                 GAAGUGAGAGUAAUUCGCCTgT 
                 21% 
                 2% 
                 16% 
                 1% 
               
               
                   
               
               
                 ND-8511 
                 325 
                 GCGAAUUACUCUCACUUCCTgT 
                 326 
                 GGAAGUGAGAGUAAUUCGCTgT 
                 12% 
                 2% 
                  6% 
                 1% 
               
               
                   
               
               
                 ND-8512 
                 327 
                 AACCAGGCGAAUUACUCUCTgT 
                 328 
                 GAGAGUAAUUCGCCUGGUUTgT 
                 99% 
                 11%  
                 79% 
                 5% 
               
               
                   
               
               
                 ND-8513 
                 329 
                 GGUAAUGGUGCACGGGCAGTgT 
                 330 
                 CUGCCCGUGCACCAUUACCTgT 
                 61% 
                 6% 
                 42% 
                 4% 
               
               
                   
               
               
                 ND-8514 
                 331 
                 CUCACGAUGGCCCUCGGUGTgT 
                 332 
                 CACCGAGGGCCAUCGUGAGTgT 
                 94% 
                 11%  
                 70% 
                 4% 
               
               
                   
               
               
                 ND-8515 
                 333 
                 GCUCCGAAGGUUCCGAAGCTgT 
                 334 
                 GCUUCGGAACCUUCGGAGCTgT 
                 18% 
                 2% 
                 17% 
                 2% 
               
               
                   
               
               
                 ND-8516 
                 335 
                 GCCGAUACUGGUCUCCAGGTgT 
                 336 
                 CCUGGAGACCAGUAUCGGCTgT 
                 14% 
                 1% 
                 18% 
                 1% 
               
               
                   
               
               
                 ND-8517 
                 337 
                 CCGAUACUGGUCUCCAGGCTgT 
                 338 
                 GCCUGGAGACCAGUAUCGGTgT 
                 42% 
                 5% 
                 33% 
                 2% 
               
               
                   
               
               
                 ND-8518 
                 339 
                 UGCUGUUGCACCAUACUUUTgT 
                 340 
                 AAAGUAUGGUGCAACAGCATgT 
                 10% 
                 1% 
                  9% 
                 0% 
               
               
                   
               
               
                 ND-8519 
                 341 
                 AACGGUCUGUCCCUGAUGCTgT 
                 342 
                 GCAUCAGGGACAGACCGUUTgT 
                 60% 
                 7% 
                 52% 
                 6% 
               
               
                   
               
               
                 ND-8520 
                 343 
                 UUAACUUGCGGCCUGGCGUTgT 
                 344 
                 ACGCCAGGCCGCAAGUUAATgT 
                 82% 
                 25%  
                 77% 
                 18%  
               
               
                   
               
               
                 ND-8521 
                 345 
                 GCUGGUUACUCACGAUGGCTgT 
                 346 
                 GCCAUCGUGAGUAACCAGCTgT 
                 36% 
                 4% 
                 34% 
                 7% 
               
               
                   
               
               
                 ND-8522 
                 347 
                 UUACUCACGAUGGCCCUCGTgT 
                 348 
                 CGAGGGCCAUCGUGAGUAATgT 
                 105%  
                 21%  
                 113%  
                 21%  
               
               
                   
               
               
                 ND-8523 
                 349 
                 GAAGCCGAUACUGGUCUCCTgT 
                 350 
                 GGAGACCAGUAUCGGCUUCTgT 
                 24% 
                 2% 
                 18% 
                 2% 
               
               
                   
               
               
                 ND-8524 
                 351 
                 GAUACUGGUCUCCAGGCCGTgT 
                 352 
                 CGGCCUGGAGACCAGUAUCTgT 
                 30% 
                 5% 
                 25% 
                 3% 
               
               
                   
               
               
                 ND-8525 
                 353 
                 AUACUAAUCUCCAGGCCGATgT 
                 354 
                 UCGGCCUGGAGACCAGUAUTgT 
                 12% 
                 1% 
                 11% 
                 2% 
               
               
                   
               
               
                 ND-8526 
                 355 
                 CAACGGUCUGUCCCUGAUGTgT 
                 356 
                 CAUCAGGGACAGACCGUUGTgT 
                 24% 
                 7% 
                 24% 
                 2% 
               
               
                   
               
               
                 ND-8527 
                 357 
                 UUUAACUUGCGGCCUGGCGTgT 
                 358 
                 CGCCAGGCCGCAAGUUAAATgT 
                 122%  
                 6% 
                 107%  
                 9% 
               
               
                   
               
               
                 ND-8528 
                 359 
                 UACUCACGAUGGCCCUCGGTgT 
                 360 
                 CCGAGGGCCAUCGUGAGUATgT 
                 78% 
                 6% 
                 84% 
                 7% 
               
               
                   
               
               
                 ND-8529 
                 361 
                 UUUCGGAGAGUACUUCAGCTgT 
                 362 
                 GCUGAAGUACUCUCCGAAATgT 
                 87% 
                 18%  
                 80% 
                 17%  
               
               
                   
               
               
                 ND-8530 
                 363 
                 GCAGACGCUCUUUGACCUGTgT 
                 364 
                 CAGGUCAAAGAGCGUCUGCTgT 
                 14% 
                 2% 
                 13% 
                 0% 
               
               
                   
               
               
                 ND-8531 
                 365 
                 CUACAUCUUCUAUCCGCGGTgT 
                 366 
                 CCGCGGAUAGAAGAUGUAGTgT 
                 20% 
                 4% 
                 18% 
                 3% 
               
               
                   
               
               
                 ND-8532 
                 367 
                 AGGCGAAUUACUCUCACUUTgT 
                 368 
                 AAGUGAGAGUAAUUCGCCUTgT 
                 25% 
                 5% 
                 18% 
                 1% 
               
               
                   
               
               
                 ND-8533 
                 369 
                 CCGCUUCAACCAGGUCUCCTgT 
                 370 
                 GGAGACCUGGUUGAAGCGGTgT 
                 30% 
                 11%  
                 22% 
                 2% 
               
               
                   
               
               
                 ND-8534 
                 371 
                 CAACCGCAUGAAGACGGCCTgT 
                 372 
                 GGCCGUCUUCAUGCGGUUGTgT 
                 33% 
                 4% 
                 23% 
                 1% 
               
               
                   
               
               
                 ND-8535 
                 373 
                 AUGAAGACGGCCUUCUGGGTgT 
                 374 
                 CCCAGAAGGCCGUCUUCAUTgT 
                 114%  
                 13%  
                 84% 
                 15%  
               
               
                   
               
               
                 ND-8536 
                 375 
                 AGCACAACCGCAUGAAGACTgT 
                 376 
                 GUCUUCAUGCGGUUGUGCUTgT 
                 18% 
                 1% 
                 16% 
                 3% 
               
               
                   
               
               
                 ND-8537 
                 377 
                 UCGAGUUCCACCGCUCCUATgT 
                 378 
                 UAGGAGCGGUGGAACUCGATgT 
                 25% 
                 0% 
                 26% 
                 3% 
               
               
                   
               
               
                 ND-8538 
                 379 
                 CUGCUUCUACCAGACAUACTgT 
                 380 
                 GUAUGUCUGGUAGAAGCAGTgT 
                 12% 
                 1% 
                 13% 
                 2% 
               
               
                   
               
               
                 ND-8539 
                 381 
                 GAGGGAGUGGUACCGCUUCTgT 
                 382 
                 GAAGCGGUACCACUCCCUCTgT 
                 43% 
                 1% 
                 47% 
                 14%  
               
               
                   
               
               
                 ND-8540 
                 383 
                 CCUUUAUGGAUGAUGGUGGTgT 
                 384 
                 CCACCAUCAUCCAUAAAGGTgT 
                 61% 
                 5% 
                 60% 
                 6% 
               
               
                   
               
               
                 ND-8541 
                 385 
                 UGAGGGAGUGGUACCGCUUTgT 
                 386 
                 AAGCGGUACCACUCCCUCATgT 
                 36% 
                 5% 
                 35% 
                 5% 
               
               
                   
               
               
                 ND-8542 
                 387 
                 CCUGCAACCAGGCGAAUUATgT 
                 388 
                 UAAUUCGCCUGGUUGCAGGTgT 
                 19% 
                 2% 
                 16% 
                 1% 
               
               
                   
               
               
                 ND-8543 
                 389 
                 GGCCUGGCGUGGAGACCUCTgT 
                 390 
                 GAGGUCUCCACGCCAGGCCTgT 
                 38% 
                 7% 
                 20% 
                 2% 
               
               
                   
               
               
                 ND-8544 
                 391 
                 UGCUUUUCGGAGAGUACUUTgT 
                 392 
                 AAGUACUCUCCGAAAAGCATgT 
                 22% 
                 5% 
                 17% 
                 1% 
               
               
                   
               
               
                 ND-8545 
                 393 
                 CCCGUAGCGUGGCCUCCAGTgT 
                 394 
                 CUGGAGGCCACGCUACGGGTgT 
                 25% 
                 3% 
                 22% 
                 2% 
               
               
                   
               
               
                 ND-8546 
                 395 
                 CCGUAGCGUGGCCUCCAGCTgT 
                 396 
                 GCUGGAGGCCACGCUACGGTgT 
                 62% 
                 5% 
                 57% 
                 9% 
               
               
                   
               
               
                 ND-8547 
                 397 
                 CCAGGCGAAUUACUCUCACTgT 
                 398 
                 GUGAGAGUAAUUCGCCUGGTgT 
                 23% 
                 11%  
                 16% 
                 2% 
               
               
                   
               
               
                 ND-8548 
                 399 
                 GAAACUGCUAUACUUUCAATgT 
                 400 
                 UUGAAAGUAUAGCAGUUUCTgT 
                  9% 
                 3% 
                  3% 
                 0% 
               
               
                   
               
               
                 ND-8549 
                 401 
                 GCCCGGGUAAUGGUGCACGTgT 
                 402 
                 CGUGCACCAUUACCCGGGCTgT 
                 87% 
                 9% 
                 92% 
                 14%  
               
               
                   
               
               
                 ND-8550 
                 403 
                 CCCGGGUAAUGGUGCACGGTgT 
                 404 
                 CCGUGCACCAUUACCCGGGTgT 
                 19% 
                 12%  
                 14% 
                 1% 
               
               
                   
               
               
                 ND-8551 
                 405 
                 CGGGUAAUGGUGCACGGGCTgT 
                 406 
                 GCCCGUGCACCAUUACCCGTgT 
                 68% 
                 11%  
                 73% 
                 3% 
               
               
                   
               
               
                 ND-8552 
                 407 
                 GGGUAAUGGUGCACGGGCATgT 
                 408 
                 UGCCCGUGCACCAUUACCCTgT 
                 30% 
                 6% 
                 33% 
                 2% 
               
               
                   
               
               
                 ND-8553 
                 409 
                 UAAUGGUGCACGGGCAGGATgT 
                 410 
                 UCCUGCCCGUGCACCAUUATgT 
                 29% 
                 3% 
                 31% 
                 1% 
               
               
                   
               
               
                 ND-8554 
                 411 
                 CUGGUUACUCACGAUGGCCTgT 
                 412 
                 GGCCAUCGUGAGUAACCAGTgT 
                 74% 
                 15%  
                 66% 
                 8% 
               
               
                   
               
               
                 ND-8555 
                 413 
                 GUUACUCACGAUGGCCCUCTgT 
                 414 
                 GAGGGCCAUCGUGAGUAACTgT 
                 91% 
                 21%  
                 88% 
                 10%  
               
               
                   
               
               
                 ND-8556 
                 415 
                 UGUCACGAUGGUCACCCUCTgT 
                 416 
                 GAGGGUGACCAUCGUGACATgT 
                 72% 
                 4% 
                 76% 
                 12%  
               
               
                   
               
               
                 ND-8557 
                 417 
                 UGCUCCGAAGGUUCCGAAGTgT 
                 418 
                 CUUCGGAACCUUCGGAGCATgT 
                 51% 
                 2% 
                 59% 
                 18%  
               
               
                   
               
               
                 ND-8558 
                 419 
                 UCCGAAGGUUCCGAAGCCGTgT 
                 420 
                 CGGCUUCGGAACCUUCGGATgT 
                 109%  
                 11%  
                 77% 
                 13%  
               
               
                   
               
               
                 ND-8559 
                 421 
                 UUCCGAAGCCGAUACUGGUTgT 
                 422 
                 ACCAGUAUCGGCUUCGGAATgT 
                 46% 
                 20%  
                 33% 
                 6% 
               
               
                   
               
               
                 ND-8560 
                 423 
                 AGCCGAUACUGGUCUCCAGTgT 
                 424 
                 CUGGAGACCAGUAUCGGCUTgT 
                 15% 
                 6% 
                 10% 
                 1% 
               
               
                   
               
               
                 ND-8561 
                 425 
                 CUUGGUACUGCCUCUGAACTgT 
                 426 
                 GUUCAGAGGCAGUACCAAGTgT 
                 16% 
                 3% 
                 12% 
                 3% 
               
               
                   
               
               
                 ND-8562 
                 427 
                 CUCCCGUAGCACACUAUAATgT 
                 428 
                 UUAUAGUGUGCUACGGGAGTgT 
                 14% 
                 6% 
                 10% 
                 1% 
               
               
                   
               
               
                 ND-8563 
                 429 
                 UCCCGUAGCACACUAUAACTgT 
                 430 
                 GUUAUAGUGUGCUACGGGATgT 
                 43% 
                 11%  
                 36% 
                 4% 
               
               
                   
               
               
                 ND-8564 
                 431 
                 UGCACCAUACUUUCUUGUATgT 
                 432 
                 UACAAGAAAGUAUGGUGCATgT 
                 17% 
                 6% 
                 13% 
                 3% 
               
               
                   
               
               
                 ND-8565 
                 433 
                 UUGCCCGUUUAUGUAUGCUTgT 
                 434 
                 AGCAUACAUAAACGGGCAATgT 
                 84% 
                 2% 
                 103%  
                 12%  
               
               
                   
               
               
                 ND-8566 
                 435 
                 UGCCCGUUUAUGUAUGCUCTgT 
                 436 
                 GAGCAUACAUAAACGGGCATgT 
                 69% 
                 25%  
                 93% 
                 4% 
               
               
                   
               
               
                 ND-8567 
                 437 
                 GGACCCUAGACCUCUGCAGTgT 
                 438 
                 CUGCAGAGGUCUAGGGUCCTgT 
                 29% 
                 8% 
                 33% 
                 2% 
               
               
                   
               
               
                 ND-8568 
                 439 
                 CCUAGACCUCUGCAGCCCATgT 
                 440 
                 UGGGCUGCAGAGGUCUAGGTgT 
                 18% 
                 2% 
                 19% 
                 1% 
               
               
                   
               
               
                 ND-8569 
                 441 
                 UGGCAUGAUGUACUGGCAATgT 
                 442 
                 UUGCCAGUACAUCAUGCCATgT 
                 19% 
                 3% 
                 20% 
                 5% 
               
               
                   
               
               
                 ND-8570 
                 443 
                 UACUGGCAAUUCGGCCUGCTgT 
                 444 
                 GCAGGCCGAAUUGCCAGUATgT 
                 86% 
                 15%  
                 83% 
                 16%  
               
               
                   
               
               
                 ND-8571 
                 445 
                 AAUUCGGCCUGCUUUUCGGTgT 
                 446 
                 CCGAAAAGCAGGCCGAAUUTgT 
                 19% 
                 3% 
                 24% 
                 4% 
               
               
                   
               
               
                 ND-8572 
                 447 
                 CUGCUUUUCGGAGAGUACUTgT 
                 448 
                 AGUACUCUCCGAAAAGCAGTgT 
                  8% 
                 2% 
                 12% 
                 2% 
               
               
                   
               
               
                 ND-8573 
                 449 
                 UUCGGAGAGUACUUCAGCUTgT 
                 450 
                 AGCUGAAGUACUCUCCGAATgT 
                 27% 
                 3% 
                 40% 
                 5% 
               
               
                   
               
               
                 ND-8574 
                 451 
                 AGCAGACGCUCUUUGACCUTgT 
                 452 
                 AGGUCAAAGAGCGUCUGCUTgT 
                 15% 
                 0% 
                 19% 
                 4% 
               
               
                   
               
               
                 ND-8575 
                 453 
                 CUUGCAGCGCCUGAGGGUCTgT 
                 454 
                 GACCCUCAGGCGCUGCAAGTgT 
                 35% 
                 1% 
                 40% 
                 4% 
               
               
                   
               
               
                 ND-8576 
                 455 
                 UGGCUUUAACUUGCGGCCUTgT 
                 456 
                 AGGCCGCAAGUUAAAGCCATgT 
                 47% 
                 3% 
                 53% 
                 8% 
               
               
                   
               
               
                 ND-8577 
                 457 
                 GCUUUAACUUGCGGCCUGGTgT 
                 458 
                 CCAGGCCGCAAGUUAAAGCTgT 
                 20% 
                 2% 
                 25% 
                 5% 
               
               
                   
               
               
                 ND-8578 
                 459 
                 UAACUUGCGGCCUGGCGUGTgT 
                 460 
                 CACGCCAGGCCGCAAGUUATgT 
                 75% 
                 7% 
                 82% 
                 4% 
               
               
                   
               
               
                 ND-8579 
                 461 
                 ACCUUUACCCUUCAAAGUATgT 
                 462 
                 UACUUUGAAGGGUAAAGGUTgT 
                 14% 
                 2% 
                 17% 
                 3% 
               
               
                   
               
               
                 ND-8580 
                 463 
                 GGUUACUCACGAUGGCCCUTgT 
                 464 
                 AGGGCCAUCGUGAGUAACCTgT 
                 63% 
                 5% 
                 70% 
                 11%  
               
               
                   
               
               
                 ND-8581 
                 465 
                 CACGAUGGCCCUCGGUGACTgT 
                 466 
                 GUCACCGAGGGCCAUCGUGTgT 
                 56% 
                 2% 
                 50% 
                 5% 
               
               
                   
               
               
                 ND-8582 
                 467 
                 AGAUGCUAUCGCGACAGAATgT 
                 468 
                 UUCUGUCGCGAUAGCAUCUTgT 
                 18% 
                 1% 
                 18% 
                 1% 
               
               
                   
               
               
                 ND-8583 
                 469 
                 ACGAUGGUCACCCUCCUGUTgT 
                 470 
                 ACAGGAGGGUGACCAUCGUTgT 
                 48% 
                 3% 
                 52% 
                 6% 
               
               
                   
               
               
                 ND-8584 
                 471 
                 CUCCGAAGGUUCCGAAGCCTgT 
                 472 
                 GGCUUCGGAACCUUCGGAGTgT 
                 18% 
                 2% 
                 28% 
                 5% 
               
               
                   
               
               
                 ND-8585 
                 473 
                 AAGGUUCCGAAGCCGAUACTgT 
                 474 
                 GUAUCGGCUUCGGAACCUUTgT 
                 26% 
                 2% 
                 28% 
                 1% 
               
               
                   
               
               
                 ND-8586 
                 475 
                 GGUACUGCCUCUGAACACUTgT 
                 476 
                 AGUGUUCAGAGGCAGUACCTgT 
                 12% 
                 1% 
                 12% 
                 1% 
               
               
                   
               
               
                 ND-8587 
                 477 
                 AGCUUUGACAAGGAACUUUTgT 
                 478 
                 AAAGUUCCUUGUCAAAGCUTgT 
                 17% 
                 2% 
                 18% 
                 2% 
               
               
                   
               
               
                 ND-8588 
                 479 
                 UUUGACAAGGAACUUUCCUTgT 
                 480 
                 AGGAAAGUUCCUUGUCAAATgT 
                 78% 
                 5% 
                 73% 
                 2% 
               
               
                   
               
               
                 ND-8589 
                 481 
                 UGACAAGGAACUUUCCUAATgT 
                 482 
                 UUAGGAAAGUUCCUUGUCATgT 
                 14% 
                 1% 
                 16% 
                 1% 
               
               
                   
               
               
                 ND-8590 
                 483 
                 CCCGUAGCACACUAUAACATgT 
                 484 
                 UGUUAUAGUGUGCUACGGGTgT 
                  9% 
                 1% 
                 11% 
                 2% 
               
               
                   
               
               
                 ND-8591 
                 485 
                 CACUAUAACAUCUGCUGGATgT 
                 486 
                 UCCAGCAGAUGUUAUAGUGTgT 
                 18% 
                 2% 
                 30% 
                 2% 
               
               
                   
               
               
                 ND-8592 
                 487 
                 UUGCUGUUGCACCAUACUUTgT 
                 488 
                 AAGUAUGGUGCAACAGCAATgT 
                 23% 
                 2% 
                 25% 
                 8% 
               
               
                   
               
               
                 ND-8593 
                 489 
                 GUACUGGCAAUUCGGCCUGTgT 
                 490 
                 CAGGCCGAAUUGCCAGUACTgT 
                 66% 
                 3% 
                 62% 
                 4% 
               
               
                   
               
               
                 ND-8594 
                 491 
                 UUCGGCCUGCUUUUCGGAGTgT 
                 492 
                 CUCCGAAAAGCAGGCCGAATgT 
                 97% 
                 7% 
                 86% 
                 8% 
               
               
                   
               
               
                 ND-8595 
                 493 
                 CCUGCUUUUCGGAGAGUACTgT 
                 494 
                 GUACUCUCCGAAAAGCAGGTgT 
                 11% 
                 3% 
                 14% 
                 3% 
               
               
                   
               
               
                 ND-8596 
                 495 
                 GCUUUUCGGAGAGUACUUCTgT 
                 496 
                 GAAGUACUCUCCGAAAAGCTgT 
                 12% 
                 1% 
                 17% 
                 2% 
               
               
                   
               
               
                 ND-8597 
                 497 
                 CUUUUCGGAGAGUACUUCATgT 
                 498 
                 UGAAGUACUCUCCGAAAAGTgT 
                 11% 
                 1% 
                 14% 
                 2% 
               
               
                   
               
               
                 ND-8598 
                 499 
                 CAACCUCAACUCGGACAAGTgT 
                 500 
                 CUUGUCCGAGUUGAGGUUGTgT 
                 15% 
                 2% 
                 16% 
                 2% 
               
               
                   
               
               
                 ND-8599 
                 501 
                 CUACCAGACAUACUCAUCATgT 
                 502 
                 UGAUGAGUAUGUCUGGUAGTgT 
                 17% 
                 1% 
                 18% 
                 2% 
               
               
                   
               
               
                 ND-8600 
                 503 
                 CUGUCGAGGCUGCCAGAGATgT 
                 504 
                 UCUCUGGCAGCCUCGACAGTgT 
                 17% 
                 0% 
                 16% 
                 1% 
               
               
                   
               
               
                 ND-8601 
                 505 
                 AAACUGCUAUACUUUCAAUTgT 
                 506 
                 AUUGAAAGUAUAGCAGUUUTgT 
                 28% 
                 1% 
                 26% 
                 1% 
               
               
                   
               
               
                 ND-8602 
                 507 
                 GGCUUUAACUUGCGGCCUGTgT 
                 508 
                 CAGGCCGCAAGUUAAAGCCTgT 
                 21% 
                 2% 
                 18% 
                 1% 
               
               
                   
               
               
                 ND-8603 
                 509 
                 CUUUAACUUGCGGCCUGGCTgT 
                 510 
                 GCCAGGCCGCAAGUUAAAGTgT 
                 81% 
                 2% 
                 69% 
                 6% 
               
               
                   
               
               
                 ND-8604 
                 511 
                 AGGUGUGUAUUCACUCCUGTgT 
                 512 
                 CAGGAGUGAAUACACACCUTgT 
                 47% 
                 4% 
                 40% 
                 1% 
               
               
                   
               
               
                 ND-8605 
                 513 
                 ACGAUGGCCCUCGGUGACATgT 
                 514 
                 UGUCACCGAGGGCCAUCGUTgT 
                 40% 
                 6% 
                 35% 
                 2% 
               
               
                   
               
               
                 ND-8606 
                 515 
                 CUGAACACUCUGGUUUCCCTgT 
                 516 
                 GGGAAACCAGAGUGUUCAGTgT 
                 60% 
                 2% 
                 75% 
                 4% 
               
               
                   
               
               
                 ND-8607 
                 517 
                 CUAUAACAUCUGCUGGAGUTgT 
                 518 
                 ACUCCAGCAGAUGUUAUAGTgT 
                 17% 
                 1% 
                 34% 
                 3% 
               
               
                   
               
               
                 ND-8608 
                 519 
                 GCACCAUACUUUCUUGUACTgT 
                 520 
                 GUACAAGAAAGUAUGGUGCTgT 
                 10% 
                 1% 
                 15% 
                 3% 
               
               
                   
               
               
                 ND-8609 
                 521 
                 UGUCUAGCCCAUCAUCCUGTgT 
                 522 
                 CAGGAUGAUGGGCUAGACATgT 
                 62% 
                 2% 
                 75% 
                 12%  
               
               
                   
               
               
                 ND-8610 
                 523 
                 AGGACCCUAGACCUCUGCATgT 
                 524 
                 UGCAGAGGUCUAGGGUCCUTgT 
                 61% 
                 5% 
                 73% 
                 10%  
               
               
                   
               
               
                 ND-8611 
                 525 
                 CCACCGCUCCUACCGAGAGTgT 
                 526 
                 CUCUCGGUAGGAGCGGUGGTgT 
                 21% 
                 2% 
                 29% 
                 5% 
               
               
                   
               
               
                 ND-8612 
                 527 
                 UACCGAGAGCUCUUCGAGUTgT 
                 528 
                 ACUCGAAGAGCUCUCGGUATgT 
                 13% 
                 1% 
                 22% 
                 3% 
               
               
                   
               
               
                 ND-8613 
                 529 
                 AACAUCCUGUCGAGGCUGCTgT 
                 530 
                 GCAGCCUCGACAGGAUGUUTgT 
                 57% 
                 2% 
                 70% 
                 4% 
               
               
                   
               
               
                 ND-8614 
                 531 
                 GAACCUUUACCCUUCAAAGTgT 
                 532 
                 CUUUGAAGGGUAAAGGUUCTgT 
                 13% 
                 3% 
                 16% 
                 2% 
               
               
                   
               
               
                 ND-8615 
                 533 
                 GGUUCCGAAGCCGAUACUGTgT 
                 534 
                 CAGUAUCGGCUUCGGAACCTgT 
                 18% 
                 1% 
                 24% 
                 2% 
               
               
                   
               
               
                 ND-8616 
                 535 
                 AAGCCGAUACUGGUCUCCATgT 
                 536 
                 UGGAGACCAGUAUCGGCUUTgT 
                 19% 
                 1% 
                 25% 
                 2% 
               
               
                   
               
               
                 ND-8617 
                 537 
                 UCUAGCCCAUCAUCCUGCUTgT 
                 538 
                 AGCAGGAUGAUGGGCUAGATgT 
                 93% 
                 3% 
                 101%  
                 3% 
               
               
                   
               
               
                 ND-8618 
                 539 
                 CGGCGCCAUCCGCCUGGUGTgT 
                 540 
                 CACCAGGCGGAUGGCGCCGTgT 
                 85% 
                 4% 
                 99% 
                 4% 
               
               
                   
               
               
                 ND-8619 
                 541 
                 UUUUCGGAGAGUACUUCAGTgT 
                 542 
                 CUGAAGUACUCUCCGAAAATgT 
                 63% 
                 2% 
                 77% 
                 3% 
               
               
                   
               
               
                 ND-8620 
                 543 
                 GAGAGUACUUCAGCUACCCTgT 
                 544 
                 GGGUAGCUGAAGUACUCUCTgT 
                 26% 
                 1% 
                 30% 
                 4% 
               
               
                   
               
               
                 ND-8621 
                 545 
                 GACGCUCUUUGACCUGUACTgT 
                 546 
                 GUACAGGUCAAAGAGCGUCTgT 
                 17% 
                 2% 
                 19% 
                 3% 
               
               
                   
               
               
                 ND-8622 
                 547 
                 UGUGUAUUCACUCCUGCUUTgT 
                 548 
                 AAGCAGGAGUGAAUACACATgT 
                 49% 
                 3% 
                 58% 
                 11%  
               
               
                   
               
               
                 ND-8623 
                 549 
                 AACAACAAGAGAAAUGGAGTgT 
                 550 
                 CUCCAUUUCUCUUGUUGUUTgT 
                 74% 
                 7% 
                 70% 
                 4% 
               
               
                   
               
               
                 ND-8624 
                 551 
                 AUUGAAGGAUGUGCAGGGCTgT 
                 552 
                 GCCCUGCACAUCCUUCAAUTgT 
                 89% 
                 6% 
                 87% 
                 12%  
               
               
                   
               
               
                 ND-8625 
                 553 
                 UCUCAGAGCCGCCCAAACUTgT 
                 554 
                 AGUUUGGGCGGCUCUGAGATgT 
                 53% 
                 3% 
                 51% 
                 6% 
               
               
                   
               
               
                 ND-8626 
                 555 
                 AAACACAACCAAGGGUACATgT 
                 556 
                 UGUACCCUUGGUUGUGUUUTgT 
                 17% 
                 2% 
                 18% 
                 2% 
               
               
                   
               
               
                 ND-8627 
                 557 
                 UACCCGUGCCCUCACAGAGTgT 
                 558 
                 CUCUGUGAGGGCACGGGUATgT 
                 58% 
                 3% 
                 55% 
                 3% 
               
               
                   
               
               
                 ND-8628 
                 559 
                 UAGCACACUAUAACAUCUGTgT 
                 560 
                 CAGAUGUUAUAGUGUGCUATgT 
                 64% 
                 3% 
                 64% 
                 15%  
               
               
                   
               
               
                 ND-8629 
                 561 
                 GGUGUGUAUUCACUCCUGCTgT 
                 562 
                 GCAGGAGUGAAUACACACCTgT 
                 25% 
                 3% 
                 23% 
                 2% 
               
               
                   
               
               
                 ND-8630 
                 563 
                 CAUGAUCAAGGAGUGUGGCTgT 
                 564 
                 GCCACACUCCUUGAUCAUGTgT 
                 32% 
                 2% 
                 26% 
                 2% 
               
               
                   
               
               
                 ND-8631 
                 565 
                 ACUCACGAUGGCCCUCGGUTgT 
                 566 
                 ACCGAGGGCCAUCGUGAGUTgT 
                 96% 
                 1% 
                 88% 
                 4% 
               
               
                   
               
               
                 ND-8632 
                 567 
                 GGAGCUUUGACAAGGAACUTgT 
                 568 
                 AGUUCCUUGUCAAAGCUCCTgT 
                 14% 
                 1% 
                 14% 
                 2% 
               
               
                   
               
               
                 ND-8633 
                 569 
                 AUACCCGUGCCCUCACAGATgT 
                 570 
                 UCUGUGAGGGCACGGGUAUTgT 
                 21% 
                 2% 
                 16% 
                 1% 
               
               
                   
               
               
                 ND-8634 
                 571 
                 GGAGUGGCCAAAGUCAACATgT 
                 572 
                 UGUUGACUUUGGCCACUCCTgT 
                 21% 
                 3% 
                 16% 
                 1% 
               
               
                   
               
               
                 ND-8635 
                 573 
                 AACUACAAAACCAAUUCUGTgT 
                 574 
                 CAGAAUUGGUUUUGUAGUUTgT 
                 49% 
                 5% 
                 37% 
                 3% 
               
               
                   
               
               
                 ND-8636 
                 575 
                 UGCUGGAGUGUUGCUGUUGTgT 
                 576 
                 CAACAGCAACACUCCAGCATgT 
                 27% 
                 3% 
                 21% 
                 2% 
               
               
                   
               
               
                 ND-8637 
                 577 
                 AGGUCUCCUGCAACCAGGCTgT 
                 578 
                 GCCUGGUUGCAGGAGACCUTgT 
                 62% 
                 8% 
                 61% 
                 4% 
               
               
                   
               
               
                 ND-8638 
                 579 
                 CUUUGGCAUGAUGUACUGGTgT 
                 580 
                 CCAGUACAUCAUGCCAAAGTgT 
                 66% 
                 6% 
                 52% 
                 8% 
               
               
                   
               
               
                 ND-8639 
                 581 
                 CAUCUGCACCCUCAAUCCCTgT 
                 582 
                 GGGAUUGAGGGUGCAGAUGTgT 
                 50% 
                 7% 
                 40% 
                 4% 
               
               
                   
               
               
                 ND-8640 
                 583 
                 CGACUGCACCAAGAAUGGCTgT 
                 584 
                 GCCAUUCUUGGUGCAGUCGTgT 
                 67% 
                 6% 
                 54% 
                 5% 
               
               
                   
               
               
                 ND-8641 
                 585 
                 AAAACACAACCAAGGGUACTgT 
                 586 
                 GUACCCUUGGUUGUGUUUUTgT 
                 14% 
                 2% 
                 14% 
                 1% 
               
               
                   
               
               
                 ND-8642 
                 587 
                 CAUCUGCUGGAGUGUUGCUTgT 
                 588 
                 AGCAACACUCCAGCAGAUGTgT 
                 13% 
                 2% 
                 13% 
                 1% 
               
               
                   
               
               
                 ND-8643 
                 589 
                 CCUACAUCUUCUAUCCGCGTgT 
                 590 
                 CGCGGAUAGAAGAUGUAGGTgT 
                 15% 
                 4% 
                 13% 
                 0% 
               
               
                   
               
               
                 ND-8644 
                 591 
                 GCCUACAUCUUCUAUCCGCTgT 
                 592 
                 GCGGAUAGAAGAUGUAGGCTgT 
                 14% 
                 3% 
                 11% 
                 1% 
               
               
                   
               
               
                 ND-8645 
                 593 
                 GAGUGGUACCGCUUCCACUTgT 
                 594 
                 AGUGGAAGCGGUACCACUCTgT 
                 16% 
                 0% 
                 20% 
                 1% 
               
               
                   
               
               
                 ND-8646 
                 595 
                 GGUACCGCUUCCACUACAUTgT 
                 596 
                 AUGUAGUGGAAGCGGUACCTgT 
                 12% 
                 0% 
                 14% 
                 1% 
               
               
                   
               
               
                 ND-8647 
                 597 
                 GUGGUACCGCUUCCACUACTgT 
                 598 
                 GUAGUGGAAGCGGUACCACTgT 
                 42% 
                 4% 
                 44% 
                 3% 
               
               
                   
               
               
                 ND-8648 
                 599 
                 GAAUUACUCUCACUUCCACTgT 
                 600 
                 GUGGAAGUGAGAGUAAUUCTgT 
                 10% 
                 1% 
                 11% 
                 3% 
               
               
                   
               
               
                 ND-8649 
                 601 
                 AAUUACUCUCACUUCCACCTgT 
                 602 
                 GGUGGAAGUGAGAGUAAUUTgT 
                 105%  
                 10%  
                 102%  
                 8% 
               
               
                   
               
               
                 ND-8650 
                 603 
                 UACUCUCACUUCCACCACCTgT 
                 604 
                 GGUGGUGGAAGUGAGAGUATgT 
                 55% 
                 6% 
                 54% 
                 8% 
               
               
                   
               
               
                 ND-8651 
                 605 
                 AGUGGUACCGCUUCCACUATgT 
                 606 
                 UAGUGGAAGCGGUACCACUTgT 
                 57% 
                 6% 
                 59% 
                 13%  
               
               
                   
               
               
                 ND-8652 
                 607 
                 GGGCAACUUCAUCUUCGCCTgT 
                 608 
                 GGCGAAGAUGAAGUUGCCCTgT 
                 47% 
                 12%  
                 36% 
                 7% 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 1B 
               
             
            
               
                   
               
               
                 Selected siRNAs in extended screening set (“human-only” siRNAs). A further 344 
               
               
                 iRNA sequences were identified and were designed to be fully complementary to 
               
               
                 the human alpha-ENaC sequences, according to the design criteria described in 
               
               
                 the examples section. All siRNAs listed in this screening set were modified 
               
               
                 only with a phosphorothioate linkage at the 3′-end between nucleotides 20 and 
               
               
                 21 of each strand. The percentage residual expression of alpha-ENaC in single- 
               
               
                 dose transfection assay is shown (refer to examples section for methods used). 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 1st 
                   
                 2nd 
                   
               
               
                   
                   
                   
                   
                   
                 screen 
                   
                 screen 
                   
               
               
                   
                   
                   
                   
                   
                 single 
                   
                 single 
                   
               
               
                   
                   
                   
                   
                   
                 dose @ 
                   
                 dose @ 
                   
               
               
                 Duplex 
                 Seq 
                   
                 Seq 
                   
                 50 nM in 
                   
                 50 nM in 
                   
               
               
                 ID 
                 ID 
                 Sense 
                 ID 
                 Antisense 
                 H441; MV 
                 SD 
                 H441; MV 
                 SD 
               
               
                   
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                 ND-10445 
                 609 
                 CUGCGGCUAAGUCUCUUUUTgT 
                 610 
                 AAAAGAGACUUAGCCGCAGTgT 
                 94% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10446 
                 611 
                 AUCGCGACAGAACAAUUACTgT 
                 612 
                 GUAAUUGUUCUGUCGCGAUTgT 
                 13% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10447 
                 613 
                 UCGCGACAGAACAAUUACATgT 
                 614 
                 UGUAAUUGUUCUGUCGCGATgT 
                 18% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10448 
                 615 
                 CCCGUUUAUGUAUGCUCCATgT 
                 616 
                 UGGAGCAUACAUAAACGGGTgT 
                 41% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10449 
                 617 
                 CCCGGGUAAGUAAAGGCAGTgT 
                 618 
                 CUGCCUUUACUUACCCGGGTgT 
                 23% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10450 
                 619 
                 GGUACCCGGAAAUUAAAGATgT 
                 620 
                 UCUUUAAUUUCCGGGUACCTgT 
                 14% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10451 
                 621 
                 GCUAUCGCGACAGAACAAUTgT 
                 622 
                 AUUGUUCUGUCGCGAUAGCTgT 
                 24% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10452 
                 623 
                 UAUCGCGACAGAACAAUUATgT 
                 624 
                 UAAUUGUUCUGUCGCGAUATgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10453 
                 625 
                 UGCGGCUAAGUCUCUUUUUTgT 
                 626 
                 AAAAAGAGACUUAGCCGCATgT 
                 46% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10454 
                 627 
                 GGCGAUUAUGGCGACUGCATgT 
                 628 
                 UGCAGUCGCCAUAAUCGCCTgT 
                 14% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10455 
                 629 
                 AUGUCUAGCCCAUCAUCCUTgT 
                 630 
                 AGGAUGAUGGGCUAGACAUTgT 
                 12% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10456 
                 631 
                 CUACAGGUACCCGGAAAUUTgT 
                 632 
                 AAUUUCCGGGUACCUGUAGTgT 
                 28% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10457 
                 633 
                 CCGUCGAGCCCGUAGCGUGTgT 
                 634 
                 CACGCUACGGGCUCGACGGTgT 
                 27% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10458 
                 635 
                 CGCGACAGAACAAUUACACTgT 
                 636 
                 GUGUAAUUGUUCUGUCGCGTgT 
                 39% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10459 
                 637 
                 AGGUACCCGGAAAUUAAAGTgT 
                 638 
                 CUUUAAUUUCCGGGUACCUTgT 
                 30% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10460 
                 639 
                 CAUGCACGGGUUUCCUGCCTgT 
                 640 
                 GGCAGGAAACCCGUGCAUGTgT 
                 95% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10461 
                 641 
                 AGCUUGCGGGACAACAACCTgT 
                 642 
                 GGUUGUUGUCCCGCAAGCUTgT 
                 94% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10462 
                 643 
                 ACUGCGGCUAAGUCUCUUUTgT 
                 644 
                 AAAGAGACUUAGCCGCAGUTgT 
                 13% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10463 
                 645 
                 CAUCCCUUAGAACCCUGCUTgT 
                 646 
                 AGCAGGGUUCUAAGGGAUGTgT 
                 18% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10464 
                 647 
                 ACCCGGGUAAGUAAAGGCATgT 
                 648 
                 UGCCUUUACUUACCCGGGUTgT 
                 41% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10465 
                 649 
                 GAUUAUGGCGACUGCACCATgT 
                 650 
                 UGGUGCAGUCGCCAUAAUCTgT 
                 23% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10466 
                 651 
                 CUCGGACAAGCUCGUCUUCTgT 
                 652 
                 GAAGACGAGCUUGUCCGAGTgT 
                 14% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10467 
                 653 
                 GCGAUUAUGGCGACUGCACTgT 
                 654 
                 GUGCAGUCGCCAUAAUCGCTgT 
                 24% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10468 
                 655 
                 AAUUACACCGUCAACAACATgT 
                 656 
                 UGUUGUUGACGGUGUAAUUTgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10469 
                 657 
                 AACUGCCGUUGAUGUGUGGTgT 
                 658 
                 CCACACAUCAACGGCAGUUTgT 
                 46% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10470 
                 659 
                 AACUGCGGCUAAGUCUCUUTgT 
                 660 
                 AAGAGACUUAGCCGCAGUUTgT 
                 14% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10471 
                 661 
                 CCGCUGAUAACCAGGACAATgT 
                 662 
                 UUGUCCUGGUUAUCAGCGGTgT 
                 12% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10472 
                 663 
                 AAGGGUACACGCAGGCAUGTgT 
                 664 
                 CAUGCCUGCGUGUACCCUUTgT 
                 28% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10473 
                 665 
                 CCGGGUAAGUAAAGGCAGATgT 
                 666 
                 UCUGCCUUUACUUACCCGGTgT 
                 27% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10474 
                 667 
                 CCCAUACCAGGUCUCAUGGTgT 
                 668 
                 CCAUGAGACCUGGUAUGGGTgT 
                 39% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10475 
                 669 
                 AUUAUGGCGACUGCACCAATgT 
                 670 
                 UUGGUGCAGUCGCCAUAAUTgT 
                 30% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10476 
                 671 
                 AUGCACGGGUUUCCUGCCCTgT 
                 672 
                 GGGCAGGAAACCCGUGCAUTgT 
                 95% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10477 
                 673 
                 CUAGCCCUCCACAGUCCACTgT 
                 674 
                 GUGGACUGUGGAGGGCUAGTgT 
                 43% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10478 
                 675 
                 CAGGUACCCGGAAAUUAAATgT 
                 676 
                 UUUAAUUUCCGGGUACCUGTgT 
                 11% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10479 
                 677 
                 AAUACAGCUCCUUCACCACTgT 
                 678 
                 GUGGUGAAGGAGCUGUAUUTgT 
                 30% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10480 
                 679 
                 CACGGGUUUCCUGCCCAGCTgT 
                 680 
                 GCUGGGCAGGAAACCCGUGTgT 
                 19% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10481 
                 681 
                 GGACUGAAUCUUGCCCGUUTgT 
                 682 
                 AACGGGCAAGAUUCAGUCCTgT 
                 14% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10482 
                 683 
                 CGUUUAUGUAUGCUCCAUGTgT 
                 684 
                 CAUGGAGCAUACAUAAACGTgT 
                 15% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10483 
                 685 
                 GGGUACUGCUACUAUAAGCTgT 
                 686 
                 GCUUAUAGUAGCAGUACCCTgT 
                 11% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10484 
                 687 
                 UCGGUGUUGUCUGUGGUGGTgT 
                 688 
                 CCACCACAGACAACACCGATgT 
                 65% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10485 
                 689 
                 AAACUGCCGUUGAUGUGUGTgT 
                 690 
                 CACACAUCAACGGCAGUUUTgT 
                 73% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10486 
                 691 
                 GCGAAACUUGGAGCUUUGATgT 
                 692 
                 UCAAAGCUCCAAGUUUCGCTgT 
                  8% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10487 
                 693 
                 GGCCCGUCGAGCCCGUAGCTgT 
                 294 
                 GCUACGGGCUCGACGGGCCTgT 
                 26% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10488 
                 695 
                 GCGACAGAACAAUUACACCTgT 
                 296 
                 GGUGUAAUUGUUCUGUCGCTgT 
                 10% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10489 
                 697 
                 GCGACGGCUUAAGCCAGCCTgT 
                 298 
                 GGCUGGCUUAAGCCGUCGCTgT 
                 50% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10490 
                 699 
                 GACCCGGGUAAGUAAAGGCTgT 
                 700 
                 GCCUUUACUUACCCGGGUCTgT 
                 74% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10491 
                 701 
                 UUGAUCACUCCGCCUUCUCTgT 
                 702 
                 GAGAAGGCGGAGUGAUCAATgT 
                 80% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10492 
                 703 
                 UCUAGCCCUCCACAGUCCATgT 
                 704 
                 UGGACUGUGGAGGGCUAGATgT 
                 69% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10493 
                 705 
                 GUUUCACCAAGUGCCGGAATgT 
                 706 
                 UUCCGGCACUUGGUGAAACTgT 
                 23% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10494 
                 707 
                 CUCAACUCGGACAAGCUCGTgT 
                 708 
                 CGAGCUUGUCCGAGUUGAGTgT 
                 45% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10495 
                 709 
                 CAACUCGGACAAGCUCGUCTgT 
                 710 
                 GACGAGCUUGUCCGAGUUGTgT 
                 23% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10496 
                 711 
                 ACCCGGAAAUUAAAGAGGATgT 
                 712 
                 UCCUCUUUAAUUUCCGGGUTgT 
                 13% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10497 
                 713 
                 CCCGGAAAUUAAAGAGGAGTgT 
                 714 
                 CUCCUCUUUAAUUUCCGGGTgT 
                 19% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10498 
                 715 
                 CACCACUCUCGUGGCCGGCTgT 
                 716 
                 GCCGGCCACGAGAGUGGUGTgT 
                 94% 
                 11%  
                   
                   
               
               
                   
               
               
                 ND-10499 
                 717 
                 CGUCGAGCCCGUAGCGUGGTgT 
                 718 
                 CCACGCUACGGGCUCGACGTgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10500 
                 719 
                 GCUUGCGGGACAACAACCCTgT 
                 720 
                 GGGUUGUUGUCCCGCAAGCTgT 
                 49% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10501 
                 721 
                 GAAUCAACAACGGUCUGUCTgT 
                 722 
                 GACAGACCGUUGUUGAUUCTgT 
                 18% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10502 
                 723 
                 GGGCGAUUAUGGCGACUGCTgT 
                 724 
                 GCAGUCGCCAUAAUCGCCCTgT 
                  8% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10503 
                 725 
                 CGAUUAUGGCGACUGCACCTgT 
                 726 
                 GGUGCAGUCGCCAUAAUCGTgT 
                 17% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10504 
                 727 
                 UCUGCUGGUUACUCACGAUTgT 
                 728 
                 AUCGUGAGUAACCAGCAGATgT 
                 38% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10505 
                 729 
                 CUAUCGCGACAGAACAAUUTgT 
                 730 
                 AAUUGUUCUGUCGCGAUAGTgT 
                  9% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10506 
                 731 
                 CAAUUACACCGUCAACAACTgT 
                 732 
                 GUUGUUGACGGUGUAAUUGTgT 
                 11% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10507 
                 733 
                 ACCGUCAACAACAAGAGAATgT 
                 734 
                 UUCUCUUGUUGUUGACGGUTgT 
                  9% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10508 
                 735 
                 CUCCUCGGUGUUGUCUGUGTgT 
                 736 
                 CACAGACAACACCGAGGAGTgT 
                 78% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10509 
                 737 
                 GGAGGUAGCCUCCACCCUGTgT 
                 738 
                 CAGGGUGGAGGCUACCUCCTgT 
                 18% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10510 
                 739 
                 GGAGAGGUUUCUCACACCATgT 
                 740 
                 UGGUGUGAGAAACCUCUCCTgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10511 
                 741 
                 CUGCCGUUGAUGUGUGGAGTgT 
                 742 
                 CUCCACACAUCAACGGCAGTgT 
                 19% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10512 
                 743 
                 UGCCGUUGAUGUGUGGAGGTgT 
                 744 
                 CCUCCACACAUCAACGGCATgT 
                 82% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10513 
                 745 
                 AGAUGGGUAAGGGCUCAGGTgT 
                 746 
                 CCUGAGCCCUUACCCAUCUTgT 
                 24% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10514 
                 747 
                 AGAACAGUAGCUGAUGAAGTgT 
                 748 
                 CUUCAUCAGCUACUGUUCUTgT 
                 15% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10515 
                 749 
                 GCGGCUAAGUCUCUUUUUCTgT 
                 750 
                 GAAAAAGAGACUUAGCCGCTgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10516 
                 751 
                 CCUAAGAAACCGCUGAUAATgT 
                 752 
                 UUAUCAGCGGUUUCUUAGGTgT 
                  6% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10517 
                 753 
                 GAAACCGCUGAUAACCAGGTgT 
                 754 
                 CCUGGUUAUCAGCGGUUUCTgT 
                 13% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10518 
                 755 
                 AACCGCUGAUAACCAGGACTgT 
                 756 
                 GUCCUGGUUAUCAGCGGUUTgT 
                 42% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10519 
                 757 
                 ACCGCUGAUAACCAGGACATgT 
                 758 
                 UGUCCUGGUUAUCAGCGGUTgT 
                 11% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10520 
                 579 
                 CCAAGGGUACACGCAGGCATgT 
                 760 
                 UGCCUGCGUGUACCCUUGGTgT 
                 19% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10521 
                 761 
                 CAAGGGUACACGCAGGCAUTgT 
                 762 
                 AUGCCUGCGUGUACCCUUGTgT 
                 12% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10522 
                 763 
                 AGGGUACACGCAGGCAUGCTgT 
                 764 
                 GCAUGCCUGCGUGUACCCUTgT 
                 23% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10523 
                 765 
                 GUACACGCAGGCAUGCACGTgT 
                 766 
                 CGUGCAUGCCUGCGUGUACTgT 
                 27% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10524 
                 767 
                 AGGCAUGCACGGGUUUCCUTgT 
                 768 
                 AGGAAACCCGUGCAUGCCUTgT 
                 14% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10525 
                 769 
                 GGCAUGCACGGGUUUCCUGTgT 
                 770 
                 CAGGAAACCCGUGCAUGCCTgT 
                 18% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10526 
                 771 
                 ACGGGUUUCCUGCCCAGCGTgT 
                 772 
                 CGCUGGGCAGGAAACCCGUTgT 
                 30% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10527 
                 773 
                 GAGCAGACCCGGGUAAGUATgT 
                 774 
                 UACUUACCCGGGUCUGCUCTgT 
                 24% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10528 
                 775 
                 AGCAGACCCGGGUAAGUAATgT 
                 776 
                 UUACUUACCCGGGUCUGCUTgT 
                 24% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10529 
                 777 
                 GGGUAAGUAAAGGCAGACCTgT 
                 778 
                 GGUCUGCCUUUACUUACCCTgT 
                 39% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10530 
                 779 
                 AGCCUCAUACCCGUGCCCUTgT 
                 780 
                 AGGGCACGGGUAUGAGGCUTgT 
                 82% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10531 
                 781 
                 GUGAACGCUUCUGCCACAUTgT 
                 782 
                 AUGUGGCAGAAGCGUUCACTgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10532 
                 783 
                 AAAUUGAUCACUCCGCCUUTgT 
                 784 
                 AAGGCGGAGUGAUCAAUUUTgT 
                 18% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10533 
                 785 
                 AAUUGAUCACUCCGCCUUCTgT 
                 786 
                 GAAGGCGGAGUGAUCAAUUTgT 
                 19% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10534 
                 787 
                 GCCUUGCGGUCAGGGACUGTgT 
                 788 
                 CAGUCCCUGACCGCAAGGCTgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10535 
                 789 
                 CUUGCGGUCAGGGACUGAATgT 
                 790 
                 UUCAGUCCCUGACCGCAAGTgT 
                 11% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10536 
                 791 
                 UUGCGGUCAGGGACUGAAUTgT 
                 792 
                 AUUCAGUCCCUGACCGCAATgT 
                 12% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10537 
                 793 
                 AUGUAUGCUCCAUGUCUAGTgT 
                 794 
                 CUAGACAUGGAGCAUACAUTgT 
                 21% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10538 
                 795 
                 AGCAAGUAGGCAGGAGCUCTgT 
                 796 
                 GAGCUCCUGCCUACUUGCUTgT 
                 19% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10539 
                 797 
                 CAGCCCAUACCAGGUCUCATgT 
                 798 
                 UGAGACCUGGUAUGGGCUGTgT 
                 27% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10540 
                 799 
                 CAGCCGUCGCGACCUGCGGTgT 
                 800 
                 CCGCAGGUCGCGACGGCUGTgT 
                 44% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10541 
                 801 
                 GGGCCCGUCGAGCCCGUAGTgT 
                 802 
                 CUACGGGCUCGACGGGCCCTgT 
                 71% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10542 
                 803 
                 CGUAGCGUGGCCUCCAGCUTgT 
                 804 
                 AGCUGGAGGCCACGCUACGTgT 
                 84% 
                 9% 
                   
                   
               
               
                   
               
               
                 ND-10543 
                 805 
                 GGUGAGGGAGUGGUACCGCTgT 
                 806 
                 GCGGUACCACUCCCUCACCTgT 
                 108%  
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10544 
                 807 
                 AAAGUACACACAGCAGGUGTgT 
                 808 
                 CACCUGCUGUGUGUACUUUTgT 
                 140%  
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10545 
                 809 
                 CCAGGUUGACUUCUCCUCATgT 
                 810 
                 UGAGGAGAAGUCAACCUGGTgT 
                 18% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10546 
                 811 
                 UGUUUCACCAAGUGCCGGATgT 
                 812 
                 UCCGGCACUUGGUGAAACATgT 
                 31% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10547 
                 813 
                 UGCUGGUUACUCACGAUGGTgT 
                 814 
                 CCAUCGUGAGUAACCAGCATgT 
                 144%  
                 10%  
                   
                   
               
               
                   
               
               
                 ND-10548 
                 815 
                 UCCUCGGUGUUGUCUGUGGTgT 
                 816 
                 CCACAGACAACACCGAGGATgT 
                 106%  
                 14%  
                   
                   
               
               
                   
               
               
                 ND-10549 
                 817 
                 AGGUAGCCUCCACCCUGGCTgT 
                 818 
                 GCCAGGGUGGAGGCUACCUTgT 
                 74% 
                 15%  
                   
                   
               
               
                   
               
               
                 ND-10550 
                 819 
                 GCCGUUGAUGUGUGGAGGGTgT 
                 820 
                 CCCUCCACACAUCAACGGCTgT 
                 26% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10551 
                 821 
                 GAUGGGUAAGGGCUCAGGATgT 
                 822 
                 UCCUGAGCCCUUACCCAUCTgT 
                 22% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10552 
                 823 
                 CCCAACUGCGGCUAAGUCUTgT 
                 824 
                 AGACUUAGCCGCAGUUGGGTgT 
                 18% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10553 
                 825 
                 CCAAGCGAAACUUGGAGCUTgT 
                 826 
                 AGCUCCAAGUUUCGCUUGGTgT 
                 16% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10554 
                 827 
                 GGGUACACGCAGGCAUGCATgT 
                 828 
                 UGCAUGCCUGCGUGUACCCTgT 
                 19% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10555 
                 829 
                 UGCACGGGUUUCCUGCCCATgT 
                 830 
                 UGGGCAGGAAACCCGUGCATgT 
                 28% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10556 
                 831 
                 CUCCUCUAGCCUCAUACCCTgT 
                 832 
                 GGGUAUGAGGCUAGAGGAGTgT 
                 109%  
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10557 
                 833 
                 UCCUCUAGCCUCAUACCCGTgT 
                 834 
                 CGGGUAUGAGGCUAGAGGATgT 
                 117%  
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10558 
                 835 
                 UCUAGCCUCAUACCCGUGCTgT 
                 836 
                 GCACGGGUAUGAGGCUAGATgT 
                 128%  
                 9% 
                   
                   
               
               
                   
               
               
                 ND-10559 
                 837 
                 UUCAUACCUCUACAUGUCUTgT 
                 838 
                 AGACAUGUAGAGGUAUGAATgT 
                 52% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10560 
                 839 
                 UCUACAUGUCUGCUUGAGATgT 
                 840 
                 UCUCAAGCAGACAUGUAGATgT 
                 15% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10561 
                 841 
                 AUAUUUCCUCAGCCUGAAATgT 
                 842 
                 UUUCAGGCUGAGGAAAUAUTgT 
                 15% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10562 
                 843 
                 AACUCCUAUGCAUCCCUUATgT 
                 844 
                 UAAGGGAUGCAUAGGAGUUTgT 
                 14% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10563 
                 845 
                 GCAUCCCUUAGAACCCUGCTgT 
                 846 
                 GCAGGGUUCUAAGGGAUGCTgT 
                 20% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10564 
                 847 
                 UGAUCACUCCGCCUUCUCCTgT 
                 848 
                 GGAGAAGGCGGAGUGAUCATgT 
                 67% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10565 
                 849 
                 UGUAAGUGCCUUGCGGUCATgT 
                 850 
                 UGACCGCAAGGCACUUACATgT 
                 17% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10566 
                 851 
                 CCUUGCGGUCAGGGACUGATgT 
                 852 
                 UCAGUCCCUGACCGCAAGGTgT 
                 14% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10567 
                 853 
                 AAUCUUGCCCGUUUAUGUATgT 
                 854 
                 UACAUAAACGGGCAAGAUUTgT 
                 13% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10568 
                 855 
                 CCGUUUAUGUAUGCUCCAUTgT 
                 856 
                 AUGGAGCAUACAUAAACGGTgT 
                 19% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10569 
                 857 
                 UGUAUGCUCCAUGUCUAGCTgT 
                 858 
                 GCUAGACAUGGAGCAUACATgT 
                 87% 
                 13%  
                   
                   
               
               
                   
               
               
                 ND-10570 
                 859 
                 CAUGUCUAGCCCAUCAUCCTgT 
                 860 
                 GGAUGAUGGGCUAGACAUGTgT 
                 33% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10571 
                 861 
                 AGUAGGCAGGAGCUCAAUATgT 
                 862 
                 UAUUGAGCUCCUGCCUACUTgT 
                 11% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10572 
                 863 
                 CCUACAGGUACCCGGAAAUTgT 
                 864 
                 AUUUCCGGGUACCUGUAGGTgT 
                 22% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10573 
                 865 
                 CCCGUCGAGCCCGUAGCGUTgT 
                 866 
                 ACGCUACGGGCUCGACGGGTgT 
                 23% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10574 
                 867 
                 GCGGUGAGGGAGUGGUACCTgT 
                 868 
                 GGUACCACUCCCUCACCGCTgT 
                 30% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10575 
                 869 
                 UUAUGGCGACUGCACCAAGTgT 
                 870 
                 CUUGGUGCAGUCGCCAUAATgT 
                 77% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10576 
                 871 
                 CUAUAAGCUCCAGGUUGACTgT 
                 872 
                 GUCAACCUGGAGCUUAUAGTgT 
                 11% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10577 
                 873 
                 UAUAAGCUCCAGGUUGACUTgT 
                 874 
                 AGUCAACCUGGAGCUUAUATgT 
                 42% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10578 
                 875 
                 AGGUUGACUUCUCCUCAGATgT 
                 876 
                 UCUGAGGAGAAGUCAACCUTgT 
                 13% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10579 
                 877 
                 CUGGGCUGUUUCACCAAGUTgT 
                 878 
                 ACUUGGUGAAACAGCCCAGTgT 
                 19% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10580 
                 879 
                 AACAAUUACACCGUCAACATgT 
                 880 
                 UGUUGACGGUGUAAUUGUUTgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10581 
                 881 
                 UGGGUAAGGGCUCAGGAAGTgT 
                 882 
                 CUUCCUGAGCCCUUACCCATgT 
                 20% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10582 
                 883 
                 GGGUAAGGGCUCAGGAAGUTgT 
                 884 
                 ACUUCCUGAGCCCUUACCCTgT 
                 22% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10583 
                 885 
                 CACCCAACUGCGGCUAAGUTgT 
                 886 
                 ACUUAGCCGCAGUUGGGUGTgT 
                 22% 
                 10%  
                   
                   
               
               
                   
               
               
                 ND-10584 
                 887 
                 ACCCAACUGCGGCUAAGUCTgT 
                 888 
                 GACUUAGCCGCAGUUGGGUTgT 
                 22% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10585 
                 889 
                 CCAACUGCGGCUAAGUCUCTgT 
                 890 
                 GAGACUUAGCCGCAGUUGGTgT 
                 14% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10586 
                 891 
                 CUUGGAUCAGCCAAGCGAATgT 
                 892 
                 UUCGCUUGGCUGAUCCAAGTgT 
                 15% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10587 
                 893 
                 GCCAAGCGAAACUUGGAGCTgT 
                 894 
                 CGUCCAAGUUUCGCUUGGCTgT 
                 17% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10588 
                 895 
                 UCCUAAGAAACCGCUGAUATgT 
                 896 
                 UAUCAGCGGUUUCUUAGGATgT 
                 11% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10589 
                 897 
                 GCAUGCACGGGUUUCCUGCTgT 
                 898 
                 GCAGGAAACCCGUGCAUGCTgT 
                 24% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10590 
                 899 
                 UGUUACUUAGGCAAUUCCCTgT 
                 900 
                 GGGAAUUGCCUAAGUAACATgT 
                 48% 
                 10%  
                   
                   
               
               
                   
               
               
                 ND-10591 
                 901 
                 CUAGGGCUAGAGCAGACCCTgT 
                 902 
                 GGGUCUGCUCUAGCCCUAGTgT 
                 58% 
                 10%  
                   
                   
               
               
                   
               
               
                 ND-10592 
                 903 
                 CUCUAGCCUCAUACCCGUGTgT 
                 904 
                 CACGGGUAUGAGGCUAGAGTgT 
                 34% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10593 
                 905 
                 UUAGAACCCUGCUCAGACATgT 
                 906 
                 UGUCUGAGCAGGGUUCUAATgT 
                 14% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10594 
                 907 
                 UGUGAACGCUUCUGCCACATgT 
                 908 
                 UGUGGCAGAAGCGUUCACATgT 
                 15% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10595 
                 909 
                 AUUGAUCACUCCGCCUUCUTgT 
                 910 
                 AGAAGGCGGAGUGAUCAAUTgT 
                 43% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10596 
                 911 
                 UCACUCCGCCUUCUCCUGGTgT 
                 912 
                 CCAGGAGAAGGCGGAGUGATgT 
                 90% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10597 
                 913 
                 GCGGUCAGGGACUGAAUCUTgT 
                 914 
                 AGAUUCAGUCCCUGACCGCTgT 
                 11% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10598 
                 915 
                 GGUCAGGGACUGAAUCUUGTgT 
                 916 
                 CAAGAUUCAGUCCCUGACCTgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10599 
                 917 
                 GUAUGCUCCAUGUCUAGCCTgT 
                 918 
                 GGCUAGACAUGGAGCAUACTgT 
                 28% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10600 
                 919 
                 CCAUGUCUAGCCCAUCAUCTgT 
                 920 
                 GAUGAUGGGCUAGACAUGGTgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10601 
                 921 
                 GAUCGAGUUCCACCGCUCCTgT 
                 922 
                 GGAGCGGUGGAACUCGAUCTgT 
                 17% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10602 
                 923 
                 GGACUCUAGCCCUCCACAGTgT 
                 924 
                 CUGUGGAGGGCUAGAGUCCTgT 
                 41% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10603 
                 925 
                 UCACCACUCUCGUGGCCGGTgT 
                 926 
                 CCGGCCACGAGAGUGGUGATgT 
                 83% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10604 
                 927 
                 CAGCUUGCGGGACAACAACTgT 
                 928 
                 GUUGUUGUCCCGCAAGCUGTgT 
                 21% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10605 
                 929 
                 CAUCUUCUAUCCGCGGCCCTgT 
                 930 
                 GGGCCGCGGAUAGAAGAUGTgT 
                 26% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10606 
                 931 
                 AUAAGCUCCAGGUUGACUUTgT 
                 932 
                 AAGUCAACCUGGAGCUUAUTgT 
                 15% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10607 
                 933 
                 CUGCUGGUUACUCACGAUGTgT 
                 934 
                 CAUCGUGAGUAACCAGCAGTgT 
                 85% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10608 
                 935 
                 GAACAAUUACACCGUCAACTgT 
                 936 
                 GUUGACGGUGUAAUUGUUCTgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10609 
                 937 
                 AUUACACCGUCAACAACAATgT 
                 938 
                 UUGUUGUUGACGGUGUAAUTgT 
                 12% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10610 
                 939 
                 CUGUGGUUCGGCUCCUCGGTgT 
                 940 
                 CCGAGGAGCCGAACCACAGTgT 
                 53% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10611 
                 941 
                 GAAGUGCCUUGGCUCCAGCTgT 
                 942 
                 GCUGGAGCCAAGGCACUUCTgT 
                 24% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10612 
                 943 
                 GAUCAGCCAAGCGAAACUUTgT 
                 944 
                 AAGUUUCGCUUGGCUGAUCTgT 
                 12% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10613 
                 945 
                 AGAAACCGCUGAUAACCAGTgT 
                 946 
                 CUGGUUAUCAGCGGUUUCUTgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10614 
                 947 
                 UGAUAACCAGGACAAAACATgT 
                 948 
                 UGUUUUGUCCUGGUUAUCATgT 
                  7% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10615 
                 949 
                 CACGCAGGCAUGCACGGGUTgT 
                 950 
                 ACCCGUGCAUGCCUGCGUGTgT 
                 12% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10616 
                 951 
                 GCUCUCCAGUAGCACAGAUTgT 
                 952 
                 AUCUGUGCUACUGGAGAGCTgT 
                  9% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10617 
                 953 
                 CAGACCCGGGUAAGUAAAGTgT 
                 954 
                 CUUUACUUACCCGGGUCUGTgT 
                 55% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10618 
                 955 
                 AGACCCGGGUAAGUAAAGGTgT 
                 956 
                 CCUUUACUUACCCGGGUCUTgT 
                 72% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10619 
                 957 
                 AUCACUCCGCCUUCUCCUGTgT 
                 958 
                 CAGGAGAAGGCGGAGUGAUTgT 
                 63% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10620 
                 959 
                 CACUCCGCCUUCUCCUGGGTgT 
                 960 
                 CCCAGGAGAAGGCGGAGUGTgT 
                 28% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10621 
                 961 
                 AACUAGACUGUAAGUGCCUTgT 
                 962 
                 AGGCACUUACAGUCUAGUUTgT 
                 23% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10622 
                 963 
                 UAUGCUCCAUGUCUAGCCCTgT 
                 964 
                 GGGCUAGACAUGGAGCAUATgT 
                 98% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10623 
                 965 
                 CCCGAUGUAUGGAAACUGCTgT 
                 966 
                 GCAGUUUCCAUACAUCGGGTgT 
                 11% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10624 
                 967 
                 GUACUGCUACUAUAAGCUCTgT 
                 968 
                 GAGCUUAUAGUAGCAGUACTgT 
                 19% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10625 
                 969 
                 AGCGUGACCAGCUACCAGCTgT 
                 970 
                 GCUGGUAGCUGGUCACGCUTgT 
                 49% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10626 
                 971 
                 ACAAUUACACCGUCAACAATgT 
                 972 
                 UUGUUGACGGUGUAAUUGUTgT 
                  8% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10627 
                 973 
                 AUGCUCCUCUGGUGGGAGGTgT 
                 974 
                 CCUCCCACCAGAGGAGCAUTgT 
                 76% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10628 
                 975 
                 AACAGUAGCUGAUGAAGCUTgT 
                 976 
                 AGCUUCAUCAGCUACUGUUTgT 
                 22% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10629 
                 977 
                 CUGACUCCCGAGGGCUAGGTgT 
                 978 
                 CCUAGCCCUCGGGAGUCAGTgT 
                 34% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10630 
                 979 
                 GUGCAACCAGAACAAAUCGTgT 
                 980 
                 CGAUUUGUUCUGGUUGCACTgT 
                 10% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10631 
                 981 
                 UGCAACCAGAACAAAUCGGTgT 
                 982 
                 CCGAUUUGUUCUGGUUGCATgT 
                 48% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10632 
                 983 
                 CUUCAAAGUACACACAGCATgT 
                 984 
                 UGCUGUGUGUACUUUGAAGTgT 
                 20% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10633 
                 985 
                 CAGCGUGACCAGCUACCAGTgT 
                 986 
                 CUGGUAGCUGGUCACGCUCTgT 
                 35% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10634 
                 987 
                 AGAACAAUUACACCGUCAATgT 
                 988 
                 UUGACGGUGUAAUUGUUCUTgT 
                 14% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10635 
                 989 
                 GAUAACCAGGACAAAACACTgT 
                 990 
                 GUGUUUUGUCCUGGUUAUCTgT 
                 11% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10636 
                 991 
                 ACAACCAAGGGUACACGCATgT 
                 992 
                 UGCGUGUACCCUUGGUUGUTgT 
                 17% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10637 
                 993 
                 CCCAGCGACGGCUUAAGCCTgT 
                 994 
                 GGCUUAAGCCGUCGCUGGGTgT 
                 27% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10638 
                 995 
                 CUCCCGAGGGCUAGGGCUATgT 
                 996 
                 UAGCCCUAGCCCUCGGGAGTgT 
                 23% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10639 
                 997 
                 UAGAACCCUGCUCAGACACTgT 
                 998 
                 GUGUCUGAGCAGGGUUCUATgT 
                 35% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10640 
                 999 
                 CCUGGGCUGUUUCACCAAGTgT 
                 1000 
                 CUUGGUGAAACAGCCCAGGTgT 
                 14% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10641 
                 1001 
                 GGAUCAGCCAAGCGAAACUTgT 
                 1002 
                 AGUUUCGCUUGGCUGAUCCTgT 
                 16% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10642 
                 1003 
                 AAGAAACCGCUGAUAACCATgT 
                 1004 
                 UGGUUAUCAGCGGUUUCUUTgT 
                 17% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10643 
                 1005 
                 ACCAAGGGUACACGCAGGCTgT 
                 1006 
                 GCCUGCGUGUACCCUUGGUTgT 
                 37% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10644 
                 1007 
                 GUAGCACAGAUGUCUGCUCTgT 
                 1008 
                 GAGCAGACAUCUGUGCUACTgT 
                 13% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10645 
                 1009 
                 UUUCAUACCUCUACAUGUCTgT 
                 1010 
                 GACAUGUAGAGGUAUGAAATgT 
                 88% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10646 
                 1011 
                 CCAACCAUCUGCCAGAGAATgT 
                 1012 
                 UUCUCUGGCAGAUGGUUGGTgT 
                 16% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10647 
                 1013 
                 GUCAGGGACUGAAUCUUGCTgT 
                 1014 
                 GCAAGAUUCAGUCCCUGACTgT 
                 16% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10648 
                 1015 
                 AGCAUGAUCAAGGAGUGUGTgT 
                 1016 
                 CACACUCCUUGAUCAUGCUTgT 
                 50% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10649 
                 1017 
                 GCAGCGUGACCAGCUACCATgT 
                 1018 
                 UGGUAGCUGGUCACGCUGCTgT 
                 40% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10650 
                 1019 
                 CAGCUCUCUGCUGGUUACUTgT 
                 1020 
                 AGUAACCAGCAGAGAGCUGTgT 
                 56% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10651 
                 1021 
                 GUUCGGCUCCUCGGUGUUGTgT 
                 1022 
                 CAACACCGAGGAGCCGAACTgT 
                 68% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10652 
                 1023 
                 GCAGAUGCUCCUCUGGUGGTgT 
                 1024 
                 CCACCAGAGGAGCAUCUGCTgT 
                 26% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10653 
                 1025 
                 AGGAAGUUGCUCCAAGAACTgT 
                 1026 
                 GUUCUUGGAGCAACUUCCUTgT 
                 18% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10654 
                 1027 
                 AACGCUUCUGCCACAUCUUTgT 
                 1028 
                 AAGAUGUGGCAGGAGCGUUTgT 
                 18% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10655 
                 1029 
                 CACCUGGGCUGUUUCACCATgT 
                 1030 
                 UGGUGAAACAGCCCAGGUGTgT 
                 17% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10656 
                 1031 
                 AAGCCAUGCAGCGUGACCATgT 
                 1032 
                 UGGUCACGCUGCAUGGCUUTgT 
                 27% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10657 
                 1033 
                 CGAGGGCUAGGGCUAGAGCTgT 
                 1034 
                 GCUCUAGCCCUAGCCCUCGTgT 
                 30% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10658 
                 1035 
                 GGAAACCCUGGACAGACUUTgT 
                 1036 
                 AAGUCUGUCCAGGGUUUCCTgT 
                 14% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10659 
                 1037 
                 GUAGCUGAUGAAGCUGCCCTgT 
                 1038 
                 GGGCAGCUUCAUCAGCUACTgT 
                 19% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10660 
                 1039 
                 UCUUUUUCCCUUGGAUCAGTgT 
                 1040 
                 CUGAUCCAAGGGAAAAAGATgT 
                 88% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10661 
                 1041 
                 CUCCAGUAGCACAGAUGUCTgT 
                 1042 
                 GACAUCUGUGCUACUGGAGTgT 
                 10% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10662 
                 1043 
                 CCAAAAUUGAUCACUCCGCTgT 
                 1044 
                 GCGGAGUGAUCAAUUUUGGTgT 
                 25% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10663 
                 1045 
                 CAGACCACCUGGGCUGUUUTgT 
                 1046 
                 AAACAGCCCAGGUGGUCUGTgT 
                 24% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10664 
                 1047 
                 CCCUUCCCAACUAGACUGUTgT 
                 1048 
                 ACAGUCUAGUUGGGAAGGGTgT 
                 15% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10665 
                 1049 
                 CGCAGCCGUCGCGACCUGCTgT 
                 1050 
                 GCAGGUCGCGACGGCUGCGTgT 
                 45% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10666 
                 1051 
                 UUCUCACACCAAGGCAGAUTgT 
                 1052 
                 AUCUGCCUUGGUGUGAGAATgT 
                 25% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10667 
                 1053 
                 CACCACCAUCCACGGCGCCTgT 
                 1054 
                 GGCGCCGUGGAUGGUGGUGTgT 
                 35% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10668 
                 1055 
                 CCAUUACUUUUGUGAACGCTgT 
                 1056 
                 GCGUUCACAAAAGUAAUGGTgT 
                 19% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10669 
                 1057 
                 CCAAGAACAGUAGCUGAUGTgT 
                 1058 
                 CAUCAGCUACUGUUCUUGGTgT 
                 23% 
                 4% 
                 16% 
                 2% 
               
               
                   
               
               
                 ND-10670 
                 1059 
                 AGGAGAGGUUUCUCACACCTgT 
                 1060 
                 GGUGUGAGAAACCUCUCCUTgT 
                 18% 
                 3% 
                 17% 
                 1% 
               
               
                   
               
               
                 ND-10671 
                 1061 
                 AUCAUCCUGCUUGGAGCAATgT 
                 1062 
                 UUGCUCCAAGCAGGAUGAUTgT 
                 33% 
                 3% 
                 24% 
                 2% 
               
               
                   
               
               
                 ND-10672 
                 1063 
                 GCAUCACAGAGCAGACGCUTgT 
                 1064 
                 AGCGUCUGCUCUGUGAUGCTgT 
                 29% 
                 2% 
                 27% 
                 4% 
               
               
                   
               
               
                 ND-10673 
                 1065 
                 AGGAGGUAGCCUCCACCCUTgT 
                 1066 
                 AGGGUGGAGGCUACCUCCUTgT 
                 63% 
                 6% 
                 61% 
                 3% 
               
               
                   
               
               
                 ND-10674 
                 1067 
                 ACAACCGCAUGAAGACGGCTgT 
                 1068 
                 GCCGUCUUCAUGCGGUUGUTgT 
                 94% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10675 
                 1069 
                 GCAUGAAGACGGCCUUCUGTgT 
                 1070 
                 CAGAAGGCCGUCUUCAUGCTgT 
                 20% 
                 2% 
                 18% 
                 1% 
               
               
                   
               
               
                 ND-10676 
                 1071 
                 GUCACGAUGGUCACCCUCCTgT 
                 1072 
                 GGAGGGUGACCAUCGUGACTgT 
                 66% 
                 5% 
                 60% 
                 4% 
               
               
                   
               
               
                 ND-10677 
                 1073 
                 CCCUGCUCAGACACCAUUATgT 
                 1074 
                 UAAUGGUGUCUGAGCAGGGTgT 
                 15% 
                 3% 
                 18% 
                 1% 
               
               
                   
               
               
                 ND-10678 
                 1075 
                 UCACGAUGGUCACCCUCCUTgT 
                 1076 
                 AGGAGGGUGACCAUCGUGATgT 
                 80% 
                 6% 
                 70% 
                 4% 
               
               
                   
               
               
                 ND-10679 
                 1077 
                 UCAACCUCAACUCGGACAATgT 
                 1078 
                 UUGUCCGAGUUGAGGUUGATgT 
                 20% 
                 3% 
                 21% 
                 1% 
               
               
                   
               
               
                 ND-10680 
                 1079 
                 UGACCAGCUACCAGCUCUCTgT 
                 1080 
                 GAGAGCUGGUAGCUGGUCATgT 
                 88% 
                 22%  
                 77% 
                 5% 
               
               
                   
               
               
                 ND-10681 
                 1081 
                 GAUGGCCCUCGGUGACAUCTgT 
                 1082 
                 GAUGUCACCGAGGGCCAUCTgT 
                 88% 
                 18%  
                 60% 
                 4% 
               
               
                   
               
               
                 ND-10682 
                 1083 
                 GCUUUGACAAGGAACUUUCTgT 
                 1084 
                 GAAAGUUCCUUGUCAAAGCTgT 
                 19% 
                 7% 
                 14% 
                 2% 
               
               
                   
               
               
                 ND-10683 
                 1085 
                 CGAUACUGGUCUCCAGGCCTgT 
                 1086 
                 GGCCUGGAGACCAGUAUCGTgT 
                 27% 
                 5% 
                 27% 
                 2% 
               
               
                   
               
               
                 ND-10684 
                 1087 
                 UCUGGAUGUCUUCCAUGCCTgT 
                 1088 
                 GGCAUGGAAGACAUCCAGATgT 
                 92% 
                 13%  
                 89% 
                 3% 
               
               
                   
               
               
                 ND-10685 
                 1089 
                 CAGGACCCUAGACCUCUGCTgT 
                 1090 
                 GCAGAGGUCUAGGGUCCUGTgT 
                 58% 
                 14%  
                 50% 
                 2% 
               
               
                   
               
               
                 ND-10686 
                 1091 
                 GACCCUAGACCUCUGCAGCTgT 
                 1092 
                 GCUGCAGAGGUCUAGGGUCTgT 
                 27% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10687 
                 1093 
                 ACCCUAGACCUCUGCAGCCTgT 
                 1094 
                 GGCUGCAGAGGUCUAGGGUTgT 
                 21% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10688 
                 1095 
                 CAGCCCACGGCGGAGGAGGTgT 
                 1096 
                 CCUCCUCCGCCGUGGGCUGTgT 
                 55% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10689 
                 1097 
                 CUCUUCGAGUUCUUCUGCATgT 
                 1098 
                 UGCAGAAGAACUCGAAGAGTgT 
                 13% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10690 
                 1099 
                 UUGGCAUGAUGUACUGGCATgT 
                 1100 
                 UGCCAGUACAUCAUGCCAATgT 
                 16% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10691 
                 1101 
                 GGCAUGAUGUACUGGCAAUTgT 
                 1102 
                 AUUGCCAGUACAUCAUGCCTgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10692 
                 1103 
                 UGUACUGGCAAUUCGGCCUTgT 
                 1104 
                 AGGCCGAAUUGCCAGUACATgT 
                 45% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10693 
                 1105 
                 ACUGGCAAUUCGGCCUGCUTgT 
                 1106 
                 AGCAGGCCGAAUUGCCAGUTgT 
                 38% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10694 
                 1107 
                 GGCAAUUCGGCCUGCUUUUTgT 
                 1108 
                 AAAAGCAGGCCGAAUUGCCTgT 
                 10% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10695 
                 1109 
                 CAAUUCGGCCUGCUUUUCGTgT 
                 1110 
                 CGAAAAGCAGGCCGAAUUGTgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10696 
                 1111 
                 UCGGAGAGUACUUCAGCUATgT 
                 1112 
                 UAGCUGAAGUACUCUCCGATgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10697 
                 1113 
                 CAACAUCCUGUCGAGGCUGTgT 
                 1114 
                 CAGCCUCGACAGGAUGUUGTgT 
                 35% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10698 
                 1115 
                 CAUCCUGUCGAGGCUGCCATgT 
                 1116 
                 UGGCAGCCUCGACAGGAUGTgT 
                 26% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10699 
                 1117 
                 UCCUGCAACCAGGCGAAUUTgT 
                 1118 
                 AAUUCGCCUGGUUGCAGGATgT 
                 28% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10700 
                 1119 
                 GGAAACUGCUAUACUUUCATgT 
                 1120 
                 UGAAAGUAUAGCAGUUUCCTgT 
                  7% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10701 
                 1121 
                 ACGGUCUGUCCCUGAUGCUTgT 
                 1122 
                 AGCAUCAGGGACAGACCGUTgT 
                 28% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10702 
                 1123 
                 GGUCUGUCCCUGAUGCUGCTgT 
                 1124 
                 GCAGCAUCAGGGACAGACCTgT 
                 33% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10703 
                 1125 
                 GGCCCGGGUAAUGGUGCACTgT 
                 1126 
                 GUGCACCAUUACCCGGGCCTgT 
                 47% 
                 10%  
                   
                   
               
               
                   
               
               
                 ND-10704 
                 1127 
                 CAGGAUGAACCUGCCUUUATgT 
                 1128 
                 UAAAGGCAGGUUCAUCCUGTgT 
                 59% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10705 
                 1129 
                 GAUGAACCUGCCUUUAUGGTgT 
                 1130 
                 CCAUAAAGGCAGGUUCAUCTgT 
                 77% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10706 
                 1131 
                 GGUGGCUUUAACUUGCGGCTgT 
                 1132 
                 GCCGCAAGUUAAAGCCACCTgT 
                 47% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10707 
                 1133 
                 GUGGCUUUAACUUGCGGCCTgT 
                 1134 
                 GGCCGCAAGUUAAAGCCACTgT 
                 17% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10708 
                 1135 
                 UUGCGGCCUGGCGUGGAGATgT 
                 1136 
                 UCUCCACGCCAGGCCGCAATgT 
                 52% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10709 
                 1137 
                 UGCGGCCUGGCGUGGAGACTgT 
                 1138 
                 GUCUCCACGCCAGGCCGCATgT 
                 81% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10710 
                 1139 
                 GCGGCCUGGCGUGGAGACCTgT 
                 1140 
                 GGUCUCCACGCCAGGCCGCTgT 
                 57% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10711 
                 1141 
                 CAGGUGUGUAUUCACUCCUTgT 
                 1142 
                 AGGAGUGAAUACACACCUGTgT 
                 24% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10712 
                 1143 
                 GUGUAUUCACUCCUGCUUCTgT 
                 1144 
                 GAAGCAGGAGUGAAUACACTgT 
                 20% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10713 
                 1145 
                 GGCCCUCGGUGACAUCCCATgT 
                 1146 
                 UGGGAUGUCACCGAGGGCCTgT 
                 40% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10714 
                 1147 
                 GAUGCUAUCGCGACAGAACTgT 
                 1148 
                 GUUCUGUCGCGAUAGCAUCTgT 
                 24% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10715 
                 1149 
                 ACUACAAAACCAAUUCUGATgT 
                 1150 
                 UCAGAAUUGGUUUUGUAGUTgT 
                 19% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10716 
                 1151 
                 CAAUUCUGAGUCUCCCUCUTgT 
                 1152 
                 AGAGGGAGACUCAGAAUUGTgT 
                 35% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10717 
                 1153 
                 CUCUGUCACGAUGGUCACCTgT 
                 1154 
                 GGUGACCAUCGUGACAGAGTgT 
                 41% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10718 
                 1155 
                 CUGCUCCGAAGGUUCCGAATgT 
                 1156 
                 UUCGGAACCUUCGGAGCAGTgT 
                 16% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10719 
                 1157 
                 AGGUUCCGAAGCCGAUACUTgT 
                 1158 
                 AGUAUCGGCUUCGGAACCUTgT 
                 16% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10720 
                 1159 
                 GUUCCGAAGCCGAUACUGGTgT 
                 1160 
                 CCAGUAUCGGCUUCGGAACTgT 
                 21% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10721 
                 1161 
                 CGAAGCCGAUACUGGUCUCTgT 
                 1162 
                 GAGACCAGUAUCGGCUUCGTgT 
                 16% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10722 
                 1163 
                 AAGAUUGAAGGAUGUGCAGTgT 
                 1164 
                 CUGCACAUCCUUCAAUCUUTgT 
                 25% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10723 
                 1165 
                 GAUUGAAGGAUGUGCAGGGTgT 
                 1166 
                 CCCUGCACAUCCUUCAAUCTgT 
                 26% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10724 
                 1167 
                 UGCCUCUGAACACUCUGGUTgT 
                 1168 
                 ACCAGAGUGUUCAGAGGCATgT 
                 45% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10725 
                 1169 
                 CCUCUGAACACUCUGGUUUTgT 
                 1170 
                 AAACCAGAGUGUUCAGAGGTgT 
                 15% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10726 
                 1171 
                 GACAAGGAACUUUCCUAAGTgT 
                 1172 
                 CUUAGGAAAGUUCCUUGUCTgT 
                 105%  
                 14%  
                   
                   
               
               
                   
               
               
                 ND-10727 
                 1173 
                 CAGGACAAAACACAACCAATgT 
                 1174 
                 UUGGUUGUGUUUUGUCCUGTgT 
                 32% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10728 
                 1175 
                 AACACAACCAAGGGUACACTgT 
                 1176 
                 GUGUACCCUUGGUUGUGUUTgT 
                 60% 
                 13%  
                   
                   
               
               
                   
               
               
                 ND-10729 
                 1177 
                 UUGAACUUGGGUGGGAAACTgT 
                 1178 
                 GUUUCCCACCCAAGUUCAATgT 
                 23% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10730 
                 1179 
                 UGAACUUGGGUGGGAAACCTgT 
                 1180 
                 GGUUUCCCACCCAAGUUCATgT 
                 18% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10731 
                 1181 
                 ACCCGUGCCCUCACAGAGCTgT 
                 1182 
                 GCUCUGUGAGGGCACGGGUTgT 
                 19% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10732 
                 1183 
                 ACUAUAACAUCUGCUGGAGTgT 
                 1184 
                 CUCCAGCAGAUGUUAUAGUTgT 
                 17% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10733 
                 1185 
                 AUCUGCUGGAGUGUUGCUGTgT 
                 1186 
                 CAGCAACACUCCAGCAGAUTgT 
                 119%  
                 20%  
                   
                   
               
               
                   
               
               
                 ND-10734 
                 1187 
                 CUGCUGGAGUGUUGCUGUUTgT 
                 1188 
                 AACAGCAACACUCCAGCAGTgT 
                 58% 
                 13%  
                   
                   
               
               
                   
               
               
                 ND-10735 
                 1189 
                 CUAGCCCAUCAUCCUGCUUTgT 
                 1190 
                 AAGCAGGAUGAUGGGCUAGTgT 
                 20% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10736 
                 1191 
                 CUCUGGAUGUCUUCCAUGCTgT 
                 1192 
                 GCAUGGAAGACAUCCAGAGTgT 
                 28% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10737 
                 1193 
                 AGCAGGACCCUAGACCUCUTgT 
                 1194 
                 AGAGGUCUAGGGUCCUGCUTgT 
                 36% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10738 
                 1195 
                 UUCGAGUUCUUCUGCAACATgT 
                 1196 
                 UGUUGCAGAAGAACUCGAATgT 
                 13% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10739 
                 1197 
                 CACCAUCCACGGCGCCAUCTgT 
                 1198 
                 GAUGGCGCCGUGGAUGGUGTgT 
                 13% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10740 
                 1199 
                 CCACGGCGCCAUCCGCCUGTgT 
                 1200 
                 CAGGCGGAUGGCGCCGUGGTgT 
                 44% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10741 
                 1201 
                 CAGCACAACCGCAUGAAGATgT 
                 1202 
                 UCUUCAUGCGGUUGUGCUGTgT 
                 23% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10742 
                 1203 
                 CCUUUGGCAUGAUGUACUGTgT 
                 1204 
                 CAGUACAUCAUGCCAAAGGTgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10743 
                 1205 
                 AUCCUGUCGAGGCUGCCAGTgT 
                 1206 
                 CUGGCAGCCUCGACAGGAUTgT 
                 14% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10744 
                 1207 
                 UCUCCUGCAACCAGGCGAATgT 
                 1208 
                 UUCGCCUGGUUGCAGGAGATgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10745 
                 1209 
                 UGCAACCAGGCGAAUUACUTgT 
                 1210 
                 AGUAAUUCGCCUGGUUGCATgT 
                 45% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10746 
                 1211 
                 ACCUCCAUCAGCAUGAGGATgT 
                 1212 
                 UCCUCAUGCUGAUGGAGGUTgT 
                 82% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10747 
                 1213 
                 GCGACUGCACCAAGAAUGGTgT 
                 1214 
                 CCAUUCUUGGUGCAGUCGCTgT 
                 82% 
                 19%  
                   
                   
               
               
                   
               
               
                 ND-10748 
                 1215 
                 ACCAAGAAUGGCAGUGAUGTgT 
                 1216 
                 CAUCACUGCCAUUCUUGGUTgT 
                 64% 
                 18%  
                   
                   
               
               
                   
               
               
                 ND-10749 
                 1217 
                 UGGUUACUCACGAUGGCCCTgT 
                 1218 
                 GGGCCAUCGUGAGUAACCATgT 
                 45% 
                 7% 
                   
                   
               
               
                   
               
               
                 ND-10750 
                 1219 
                 AGAAAUGGAGUGGCCAAAGTgT 
                 1220 
                 CUUUGGCCACUCCAUUUCUTgT 
                 11% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10751 
                 1221 
                 GGAGCUGAACUACAAAACCTgT 
                 1222 
                 GGUUUUGUAGUUCAGCUCCTgT 
                 15% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10752 
                 1223 
                 CCUCUGUCACGAUGGUCACTgT 
                 1224 
                 GUGACCAUCGUGACAGAGGTgT 
                 18% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10753 
                 1225 
                 AGAUUGAAGGAUGUGCAGGTgT 
                 1226 
                 CCUGCACAUCCUUCAAUCUTgT 
                 26% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10754 
                 1227 
                 GAGCUUUGACAAGGAACUUTgT 
                 1228 
                 AAGUUCCUUGUCAAAGCUCTgT 
                 14% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10755 
                 1229 
                 CUUUGACAAGGAACUUUCCTgT 
                 1230 
                 GGAAAGUUCCUUGUCAAAGTgT 
                 50% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10756 
                 1231 
                 UCAGACACCAUUACUUUUGTgT 
                 1232 
                 CAAAAGUAAUGGUGUCUGATgT 
                 32% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10757 
                 1233 
                 AGCACACUAUAACAUCUGCTgT 
                 1234 
                 GCAGAUGUUAUAGUGUGCUTgT 
                 11% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10758 
                 1235 
                 GCACAACCGCAUGAAGACGTgT 
                 1236 
                 CGUCUUCAUGCGGUUGUGCTgT 
                 34% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10759 
                 1237 
                 ACUGCUUCUACCAGACAUATgT 
                 1238 
                 UAUGUCUGGUAGAAGCAGUTgT 
                 11% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10760 
                 1239 
                 GAAGACGGCCUUCUGGGCATgT 
                 1240 
                 UGCCCAGAAGGCCGUCUUCTgT 
                 16% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10761 
                 1241 
                 AAGACGGCCUUCUGGGCAGTgT 
                 1242 
                 CUGCCCAGAAGGCCGUCUUTgT 
                 58% 
                 21%  
                   
                   
               
               
                   
               
               
                 ND-10762 
                 1243 
                 ACAUCAACCUCAACUCGGATgT 
                 1244 
                 UCCGAGUUGAGGUUGAUGUTgT 
                 14% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10763 
                 1245 
                 UGGAAGGACUGGAAGAUCGTgT 
                 1246 
                 CGAUCUUCCAGUCCUUCCATgT 
                 109%  
                 29%  
                   
                   
               
               
                   
               
               
                 ND-10764 
                 1247 
                 ACAUCCUGUCGAGGCUGCCTgT 
                 1248 
                 GGCAGCCUCGACAGGAUGUTgT 
                 101%  
                 13%  
                   
                   
               
               
                   
               
               
                 ND-10765 
                 1249 
                 CAACCAGGCGAAUUACUCUTgT 
                 1250 
                 AGAGUAAUUCGCCUGGUUGTgT 
                 19% 
                 5% 
                   
                   
               
               
                   
               
               
                 ND-10766 
                 1251 
                 CAGGCGAAUUACUCUCACUTgT 
                 1252 
                 AGUGAGAGUAAUUCGCCUGTgT 
                 24% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10767 
                 1253 
                 AGCAGAAUGACUUCAUUCCTgT 
                 1254 
                 GGAAUGAAGUCAUUCUGCUTgT 
                 40% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10768 
                 1255 
                 AUGAUGGUGGCUUUAACUUTgT 
                 1256 
                 AAGUUAAAGCCACCAUCAUTgT 
                 85% 
                 8% 
                   
                   
               
               
                   
               
               
                 ND-10769 
                 1257 
                 AGAACCUUUACCCUUCAAATgT 
                 1258 
                 UUUGAAGGGUAAAGGUUCUTgT 
                 22% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10770 
                 1259 
                 CCUUUACCCUUCAAAGUACTgT 
                 1260 
                 GUACUUUGAAGGGUAAAGGTgT 
                 21% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10771 
                 1261 
                 GAGCCUGUGGUUCGGCUCCTgT 
                 1262 
                 GGAGCCGAACCACAGGCUCTgT 
                 28% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10772 
                 1263 
                 UGGUACUGCCUCUGAACACTgT 
                 1264 
                 GUGUUCAGAGGCAGUACCATgT 
                 58% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10773 
                 1265 
                 CUCAUACCCGUGCCCUCACTgT 
                 1266 
                 GUGAGGGCACGGGUAUGAGTgT 
                 15% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10774 
                 1267 
                 CCGUAGCACACUAUAACAUTgT 
                 1268 
                 AUGUUAUAGUGUGCUACGGTgT 
                 24% 
                 6% 
                   
                   
               
               
                   
               
               
                 ND-10775 
                 1269 
                 CGUAGCACACUAUAACAUCTgT 
                 1270 
                 GAUGUUAUAGUGUGCUACGTgT 
                 21% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10776 
                 1271 
                 GCAGGACCCUAGACCUCUGTgT 
                 1272 
                 CAGAGGUCUAGGGUCCUGCTgT 
                 25% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10777 
                 1273 
                 GCCUGCUUUUCGGAGAGUATgT 
                 1274 
                 UACUCUCCGAAAAGCAGGCTgT 
                 18% 
                 4% 
                   
                   
               
               
                   
               
               
                 ND-10778 
                 1275 
                 GGGCCCGGGUAAUGGUGCATgT 
                 1276 
                 UGCACCAUUACCCGGGCCCTgT 
                 16% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10779 
                 1277 
                 CAACAACAAGAGAAAUGGATgT 
                 1278 
                 UCCAUUUCUCUUGUUGUUGTgT 
                 17% 
                 0% 
                   
                   
               
               
                   
               
               
                 ND-10780 
                 1279 
                 GCUGUUGCACCAUACUUUCTgT 
                 1280 
                 GAAAGUAUGGUGCAACAGCTgT 
                 14% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10781 
                 1281 
                 CUACCGAGAGCUCUUCGAGTgT 
                 1282 
                 CUCGAAGAGCUCUCGGUAGTgT 
                 24% 
                 3% 
                   
                   
               
               
                   
               
               
                 ND-10782 
                 1283 
                 ACCUGCCUUUAUGGAUGAUTgT 
                 1284 
                 AUCAUCCAUAAAGGCAGGUTgT 
                 115%  
                 10%  
                   
                   
               
               
                   
               
               
                 ND-10783 
                 1285 
                 UUGACAAGGAACUUUCCUATgT 
                 1286 
                 UAGGAAAGUUCCUUGUCAATgT 
                 16% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10784 
                 1287 
                 GCUGGAGUGUUGCUGUUGCTgT 
                 1288 
                 GCAACAGCAACACUCCAGCTgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10785 
                 1289 
                 UCGGUGACAUCCCAGGAAUTgT 
                 1290 
                 AUUCCUGGGAUGUCACCGATgT 
                 28% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10786 
                 1291 
                 GCUGCCCAGAAGUGCCUUGTgT 
                 1292 
                 CAAGGCACUUCUGGGCAGCTgT 
                 15% 
                 2% 
                   
                   
               
               
                   
               
               
                 ND-10787 
                 1293 
                 AGUACACACAGCAGGUGUGTgT 
                 1294 
                 CACACCUGCUGUGUGUACUTgT 
                 12% 
                 1% 
                   
                   
               
               
                   
               
               
                 ND-10788 
                 1295 
                 CAAGUGCCGGAAGCCAUGCTgT 
                 1296 
                 GCAUGGCUUCCGGCACUUGTgT 
                 94% 
                 2% 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 1C 
               
             
            
               
                   
               
               
                 Selected siRNAs in in vivo rat surrogate set (human-rat cross-reaclive 
               
               
                 siRNAs with highest specificity in rat). A screening set of 48 human and rat cross-reactive 
               
               
                 alpha-ENaC iRNA sequences were identified. The percentage residual expression of alpha- 
               
               
                 ENaC in two independent single-dose transfection experiments is shown (refer to examples 
               
               
                 section for methods used). 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 1st 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 screen 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 single 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 dose @ 
                   
                 2nd 
                   
               
               
                   
                   
                   
                   
                   
                 50 nM 
                   
                 screen 
                   
               
               
                   
                   
                   
                   
                   
                 in 
                   
                 @ 50 
                   
               
               
                 Duplex 
                   
                   
                   
                   
                 H441 
                   
                 nM in 
                   
               
               
                 ID 
                 Seq ID 
                 Sense 
                 Seq ID 
                 Antissense 
                 MV 
                 SD 
                 H441 
                 SD 
               
               
                   
               
               
                 ND-9202 
                 1297 
                 uGuGcAAccAGAAcAAAucTsT 
                 1298 
                 GAUUUGUUCUGGUUGcAcATsT 
                  8% 
                 1% 
                  8% 
                 1% 
               
               
                   
               
               
                 ND-9202 
                 2299 
                 uuuAuGGAuGAuGGuGGcuTsT 
                 1300 
                 AGCcACcAUcAUCcAuAAATsT 
                 80% 
                 9% 
                 82% 
                 6% 
               
               
                   
               
               
                 ND-9203 
                 1301 
                 GccuuuAuGGAuGAuGGuGTsT 
                 1302 
                 cACcAUcAUCcAuAAAGGCTsT 
                 76% 
                 8% 
                 76% 
                 2% 
               
               
                   
               
               
                 ND-9204 
                 2303 
                 cAcAAccGcAuGAAGAcGGTsT 
                 1304 
                 CCGUCUUcAUGCGGUUGUGTsT 
                 73% 
                 18%  
                 57% 
                 3% 
               
               
                   
               
               
                 ND-9205 
                 1305 
                 AccGcAuGAAGAcGGccuuTsT 
                 1306 
                 AAGGCCGUCUUcAUGCGGUTsT 
                 35% 
                 3% 
                 37% 
                 2% 
               
               
                   
               
               
                 ND-9206 
                 1307 
                 AGGAcuGGAAGAucGGcuuTsT 
                 1308 
                 AAGCCGAUCUUCcAGUCCUTsT 
                 17% 
                 3% 
                 16% 
                 3% 
               
               
                   
               
               
                 ND-9207 
                 1309 
                 GAAGGAcuGGAAGAucGGcTsT 
                 1310 
                 GCCGAUCUUCcAGUCCUUCTsT 
                 96% 
                 18%  
                 81% 
                 5% 
               
               
                   
               
               
                 ND-9208 
                 1311 
                 GGAcuGGAAGAuCGGcUUcTsT 
                 1312 
                 GAAGCCGAUCUUCcAGUCCTsT 
                 58% 
                 6% 
                 57% 
                 3% 
               
               
                   
               
               
                 ND-9209 
                 1313 
                 AGuuccAccGcuccuAccGTsT 
                 1314 
                 CGGuAGGAGCGGUGGAACUTsT 
                 85% 
                 8% 
                 94% 
                 4% 
               
               
                   
               
               
                 ND-9210 
                 1315 
                 GAcuGGAAGAucGGcuuccTsT 
                 1316 
                 GGAAGCCGAUCUUCcAGUCTsT 
                 79% 
                 5% 
                 82% 
                 2% 
               
               
                   
               
               
                 ND-9211 
                 1317 
                 cGcAuGAAGAcGGccuucuTsT 
                 1318 
                 AGAAGCCCGUCUUcAUGCCTsT 
                 50% 
                 1% 
                 51% 
                 1% 
               
               
                   
               
               
                 ND-9212 
                 1219 
                 GccAGuGGAGccuGuGGuuTsT 
                 1320 
                 AACcAcAGGCUCcACUGGCTsT 
                 26% 
                 3% 
                 23% 
                 2% 
               
               
                   
               
               
                 ND-9213 
                 1321 
                 uGccuuuAuGGAuGAuGGuTsT 
                 1322 
                 ACcAUcAUCcAuAAAGGcATsT 
                 77% 
                 5% 
                 76% 
                 4% 
               
               
                   
               
               
                 ND-9214 
                 1323 
                 uccuGuCCAAccuGGGcAGTsT 
                 1324 
                 CUGCCcAGGUUGGAcAGGATsT 
                 74% 
                 9% 
                 83% 
                 6% 
               
               
                   
               
               
                 ND-9215 
                 1325 
                 AGGGAGuGGuAccGcuuccTsT 
                 1326 
                 GGAAGCGGuACcACUCCCUTsT 
                 79% 
                 6% 
                 89% 
                 4% 
               
               
                   
               
               
                 ND-9216 
                 1327 
                 GGcuGuGccuAcAucuucuTsT 
                 1328 
                 AGAAGAUGuAGGcAcAGCCTsT 
                 11% 
                 1% 
                 13% 
                 1% 
               
               
                   
               
               
                 ND-9217 
                 1329 
                 GAAAuuAAAGAGGAGuuGGTsT 
                 1330 
                 CcAGCUCCUCUUuAAUUUCTsT 
                 84% 
                 14%  
                 78% 
                 5% 
               
               
                   
               
               
                 ND-9218 
                 1331 
                 AcuGGAAGAucGGcuuccATsT 
                 1332 
                 UGGAAGCCGAUCUUCcAGUTsT 
                 50% 
                 4% 
                 55% 
                 3% 
               
               
                   
               
               
                 ND-9219 
                 1333 
                 ccuGuccAAccuGGGcAGcTsT 
                 1334 
                 GCUGCCcAGGUUGGAcAGGTsT 
                 78% 
                 6% 
                 85% 
                 5% 
               
               
                   
               
               
                 ND-9220 
                 1335 
                 ccuGccuuuAuGGAuGAuGTsT 
                 1336 
                 cAUcAUCcAuAAAGGcAGGTsT 
                 76% 
                 5% 
                 77% 
                 9% 
               
               
                   
               
               
                 ND-9221 
                 1337 
                 AAccGcAuGAAGAcGGccuTsT 
                 1338 
                 AGGCCGUCUUcAUGCGGUUTsT 
                 79% 
                 8% 
                 66% 
                 3% 
               
               
                   
               
               
                 ND-9222 
                 1339 
                 uGuccAAccuGGGcAGccATsT 
                 1340 
                 UGGCUGCCcAGGUUGGAcATsT 
                 70% 
                 4% 
                 57% 
                 4% 
               
               
                   
               
               
                 ND-9223 
                 1341 
                 GuccAAccuGGGcAGccAGTsT 
                 1342 
                 CUGGCUGCCcAGGUUGGACTsT 
                 95% 
                 10%  
                 76% 
                 4% 
               
               
                   
               
               
                 ND-9224 
                 1343 
                 AAAuuAAAGAGGAGcuGGATsT 
                 1344 
                 UCcAGCUCCUCUUuAAUUUTsT 
                 83% 
                 6% 
                 69% 
                 2% 
               
               
                   
               
               
                 ND-9225 
                 1345 
                 GGAAGGAcuGGAAGAucGGTsT 
                 1346 
                 CCGAUCUUCcAGUCCUUCCTsT 
                 41% 
                 2% 
                 30% 
                 2% 
               
               
                   
               
               
                 ND-9226 
                 1347 
                 GuGAGGGAGuGGuAccGcuTsT 
                 1348 
                 AGCGGuACcACUCCCUcACTsT 
                 21% 
                 1% 
                 17% 
                 0% 
               
               
                   
               
               
                 ND-9227 
                 1349 
                 AcuuucAAuGAcAAGAAcATsT 
                 1350 
                 UGUUCUUGUcAUUGAAAGUTsT 
                 13% 
                 1% 
                 10% 
                 0% 
               
               
                   
               
               
                 ND-9228 
                 1351 
                 ucAAuGAcAAGAAcAAcucTsT 
                 1352 
                 GAGUUGUUCUUGUcAUUGATsT 
                 36% 
                 2% 
                 28% 
                 0% 
               
               
                   
               
               
                 ND-9229 
                 1353 
                 cuuuAuGGAuGAuGGuGGcTsT 
                 1354 
                 GCcACcAUcAUCcAuAAAGTsT 
                 24% 
                 1% 
                 20% 
                 1% 
               
               
                   
               
               
                 ND-9230 
                 1355 
                 GccuGGcGuGGAGAccuccTsT 
                 1356 
                 GGAGGUCUCcACGCcAGGCTsT 
                   
                   
                   
                   
               
               
                   
               
               
                 ND-9231 
                 1357 
                 uGGcGuGGAGAccuCCAuCTsT 
                 1358 
                 GAUGGAGGUCUCcACGCcATsT 
                 45% 
                 4% 
                 35% 
                 2% 
               
               
                   
               
               
                 ND-9232 
                 1359 
                 GAGuuccAccGcuccuAccTsT 
                 1360 
                 GGuAGGAGCGGUGGAACUCTsT 
                 89% 
                 4% 
                 86% 
                 8% 
               
               
                   
               
               
                 ND-9233 
                 1361 
                 cAGAGcAGAAuGAcuucAuTsT 
                 1362 
                 AUGAAGUcAUUCUGCUCUGTsT 
                 21% 
                 1% 
                 17% 
                 8% 
               
               
                   
               
               
                 ND-9234 
                 1363 
                 uucAcuccuGcuuccAGGATsT 
                 1364 
                 UCCUGGAAGcAGGAGUGAATsT 
                 85% 
                 4% 
                 74% 
                 5% 
               
               
                   
               
               
                 ND-9235 
                 1365 
                 ucAcuccuGcuuccAGGAGTsT 
                 1366 
                 CUCCUGGAAGcAGGAGUGATsT 
                   
                   
                   
                   
               
               
                   
               
               
                 ND-9236 
                 1367 
                 cuGuGcAAccAGAAcAAAuTsT 
                 1368 
                 AUUUGUUCUGGUUGcAcAGTsT 
                 23% 
                 1% 
                 17% 
                 2% 
               
               
                   
               
               
                 ND-9237 
                 1369 
                 cuGcAAcAAcAccAccAucTsT 
                 1370 
                 GAUGGUGGUGUUGUUGcAGTsT 
                 34% 
                 2% 
                 27% 
                 2% 
               
               
                   
               
               
                 ND-9238 
                 1371 
                 uGuGGcuGuGccuAcAucuTsT 
                 1372 
                 AGAUGuAGGcAcAGCcAcATsT 
                 86% 
                 4% 
                 73% 
                 10%  
               
               
                   
               
               
                 ND-9239 
                 1373 
                 uGGcuGuGccuAcAucuucTsT 
                 1374 
                 GAAGAUGuAGGcAcAGCcATsT 
                 68% 
                 6% 
                 53% 
                 4% 
               
               
                   
               
               
                 ND-9240 
                 1375 
                 cuGuccAAccuGCGcAGccTsT 
                 1376 
                 GGCUGCCcAGGUUGGAcAGTsT 
                 80% 
                 5% 
                 73% 
                 9% 
               
               
                   
               
               
                 ND-9241 
                 1377 
                 cccuGcuGuccAcAGuGAcTsT 
                 1378 
                 GUcACUGUGGAcAGcAGGGTsT 
                 83% 
                 5% 
                 71% 
                 5% 
               
               
                   
               
               
                 ND-9242 
                 1379 
                 GcAGccAGuGGAGccuGuGTsT 
                 1380 
                 cAcAGGCUCcACUGGCUGCTsT 
                 105%  
                 9% 
                 90% 
                 5% 
               
               
                   
               
               
                 ND-9243 
                 1381 
                 uucAAuGAcAAGAAcAAcuTsT 
                 1382 
                 AGUUGUUCUUGUcAUUGAATsT 
                 23% 
                 3% 
                 21% 
                 1% 
               
               
                   
               
               
                 ND-9244 
                 1383 
                 cuGccuuuAuGGAuGAuGGTsT 
                 1384 
                 CcAUcAUCcAuAAAGGcAGTsT 
                 74% 
                 6% 
                 64% 
                 7% 
               
               
                   
               
               
                 ND-9245 
                 1385 
                 AAuGAcAAGAAcAAcuccATsT 
                 1386 
                 UGGAGUUGUUCUUGUcAUUTsT 
                 21% 
                 1% 
                 21% 
                 1% 
               
               
                   
               
               
                 ND-9246 
                 1387 
                 uGGGcAGccAGuGGAGccuTsT 
                 1388 
                 AGGCUCcACUGGCUGCCcATsT 
                 83% 
                 3% 
                 73% 
                 2% 
               
               
                   
               
               
                 ND-9247 
                 1389 
                 cuccuGuccAAccuGGGcATsT 
                 1390 
                 UGCCcAGGUUGGAcAGGAGTsT 
                 86% 
                 3% 
                 84% 
                 1% 
               
               
                   
               
               
                 ND-9248 
                 1391 
                 GGcGuGGAGAccuccAucATsT 
                 1392 
                 UGAUGGAGGUCUCcACGCCTsT 
                 92% 
                 4% 
                 88% 
                 3% 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 1D 
               
             
            
               
                   
               
               
                 Selected siRNAs in in vivo guinea pig surrogate set (human-guinea pig 
               
               
                 cross-reactive siRNAs). A screening set of 63 human and guinea pig cross-reactive alpha- 
               
               
                 -ENaC iRNA sequences were identified and synthesised, both with (sequence strands 1393- 
               
               
                 1518) and without (sequence strands 1519-1644) backbone modification. The percentage 
               
               
                 residual expression of alpha-ENaC in two independent single-dose transfection experiments 
               
               
                 is shown (refer to examples section for methods used). 
               
            
           
           
               
               
               
               
               
               
               
               
               
            
               
                   
                   
                   
                   
                   
                 1st 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 screen 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 single 
                   
                   
                   
               
               
                   
                   
                   
                   
                   
                 dose @ 
                   
                 2nd 
                   
               
               
                   
                   
                   
                   
                   
                 50 nM 
                   
                 screen 
                   
               
               
                   
                   
                   
                   
                   
                 in 
                   
                 @ 50 
                   
               
               
                 Duplex 
                 Seq 
                   
                 Seq 
                   
                 H441; 
                   
                 nM in 
                   
               
               
                 ID 
                 ID 
                 Sense 
                 ID 
                 Antissense 
                 NV 
                 SD 
                 H441 
                 SD 
               
               
                   
               
               
                 ND8437 
                 1393 
                 AAucGGAcuGcuucuAccATsT 
                 1394 
                 UGGuAGAAGcAGUCCGAUUTsT 
                 48% 
                 7% 
                 46% 
                 5% 
               
               
                   
               
               
                 ND8438 
                 1395 
                 AucGGAcuGcuucuAccAGTsT 
                 1396 
                 CUGGuAGAAGcAGUCCGAUTsT 
                 85% 
                 5% 
                 93% 
                 13%  
               
               
                   
               
               
                 ND8439 
                 1397 
                 AAAucGGAcuGcuucuAccTsT 
                 1398 
                 GGuAGAAGcAGUCCGAUUUTsT 
                 36% 
                 3% 
                 42% 
                 6% 
               
               
                   
               
               
                 ND8440 
                 1399 
                 ucGGAcuGcuucuAccAGATsT 
                 1400 
                 UCUGGuAGAAGcAGUCCGATsT 
                 45% 
                 3% 
                 50% 
                 4% 
               
               
                   
               
               
                 ND8441 
                 1401 
                 AccAGAAcAAAucGGAcuGTsT 
                 1402 
                 cAGUCCGAUUUGUUCUGGUTsT 
                 23% 
                 3% 
                 24% 
                 6% 
               
               
                   
               
               
                 ND8442 
                 1403 
                 ccAGAAcAAAucGGAcuGcTsT 
                 1404 
                 GcAGUCCGAUUUGUUCUGGTsT 
                 50% 
                 6% 
                 39% 
                 9% 
               
               
                   
               
               
                 ND8443 
                 1405 
                 cAGAAcAAAucGGAcuGcuTsT 
                 1406 
                 AGcAGUCCGAUUUGUUCUGTsT 
                 22% 
                 2% 
                 24% 
                 1% 
               
               
                   
               
               
                 ND8444 
                 1407 
                 cuucGccuGccGcuucAAcTsT 
                 1408 
                 GUUGAAGCGGcAGGCGAAGTsT 
                 111%  
                 8% 
                 109%  
                 4% 
               
               
                   
               
               
                 ND8445 
                 1409 
                 uGGuAccGcuuccAcuAcATsT 
                 1410 
                 UGuAGUGGAAGCGGuACcATsT 
                 84% 
                 7% 
                 97% 
                 13%  
               
               
                   
               
               
                 ND8446 
                 1411 
                 AucuucGccuGccGcuucATsT 
                 1412 
                 UGAAGCGGcAGGCGAAGAUTsT 
                 90% 
                 3% 
                 121%  
                 13%  
               
               
                   
               
               
                 ND8447 
                 1413 
                 uucGccuGccGcuucAAccTsT 
                 1414 
                 GGUUGAAGCGGcAGGCGAATsT 
                 92% 
                 2% 
                 105%  
                 17%  
               
               
                   
               
               
                 ND8448 
                 1415 
                 cAcccucAAucccuAcAGGTsT 
                 1416 
                 CCUGuAGGGAUUGAGGGUGTsT 
                 79% 
                 9% 
                 90% 
                 13%  
               
               
                   
               
               
                 ND8449 
                 1417 
                 AGAAcAAAuCGGAcuGcuuTsT 
                 1418 
                 AAGcAGUCCGAUUUGUUCUTsT 
                 11% 
                 8% 
                 17% 
                 3% 
               
               
                   
               
               
                 ND8450 
                 1419 
                 GAAcAAAuCGGAcuGcuucTsT 
                 1420 
                 GAAGcAGUCCGAUUUGUUCTsT 
                 21% 
                 1% 
                 30% 
                 5% 
               
               
                   
               
               
                 ND8451 
                 1421 
                 cGGAcuGcuucuAccAGAcTsT 
                 1422 
                 GUCUGGuAGAAGcAGUCCGTsT 
                 24% 
                 2% 
                 32% 
                 5% 
               
               
                   
               
               
                 ND8452 
                 1423 
                 AGccucAAcAucAAccucATsT 
                 1424 
                 UGAGGUUGAUGUUGAGGCUTsT 
                 51% 
                 3% 
                 57% 
                 4% 
               
               
                   
               
               
                 ND8453 
                 1425 
                 GccucAAcAucAAccucAATsT 
                 1426 
                 UUGAGGUUGAUGUUGAGGCTsT 
                 16% 
                 1% 
                 26% 
                 3% 
               
               
                   
               
               
                 ND8454 
                 1427 
                 GucAGccucAAcAucAAccTsT 
                 1428 
                 GGUUGAUGUUGAGGCUGACTsT 
                 62% 
                 5% 
                 68% 
                 6% 
               
               
                   
               
               
                 ND8455 
                 1429 
                 ucAGccucAAcAucAAccuTsT 
                 1430 
                 AGGUUGAUGUUGAGGCUGATsT 
                 77% 
                 4% 
                 87% 
                 6% 
               
               
                   
               
               
                 ND8456 
                 1431 
                 cAGccucAAcAucAAccucTsT 
                 1432 
                 GAGGUUGAUGUUGAGGCUGTsT 
                 34% 
                 2% 
                 51% 
                 8% 
               
               
                   
               
               
                 ND8457 
                 1433 
                 GGAGcuGGAccGcAucAcATsT 
                 1434 
                 UGUGAUGCGGUCcAGCUCCTsT 
                 26% 
                 2% 
                 17% 
                 1% 
               
               
                   
               
               
                 ND8458 
                 1435 
                 GuAccGcuuccAcuAcAucTsT 
                 1436 
                 GAUGuAGUGGAAGCGGuACTsT 
                 101%  
                 9% 
                 99% 
                 11%  
               
               
                   
               
               
                 ND8459 
                 1437 
                 ccGcuuccAcuAcAucAAcTsT 
                 1438 
                 GUUGAUGuAGUGGAAGCGGTsT 
                 85% 
                 8% 
                 80% 
                 6% 
               
               
                   
               
               
                 ND8460 
                 1439 
                 cGcuuccAcuAcAucAAcATsT 
                 1440 
                 UGUUGAUGuAGUGGAAGCGTsT 
                 56% 
                 6% 
                 48% 
                 3% 
               
               
                   
               
               
                 ND8461 
                 1441 
                 uuccAcuAcAucAAcAuccTsT 
                 1442 
                 GGAUGUUGAUGuAGUGGAATsT 
                 77% 
                 5% 
                 82% 
                 7% 
               
               
                   
               
               
                 ND8462 
                 1443 
                 uGGGcAAcuucAucuucGcTsT 
                 1444 
                 GCGAAGAUGAAGUUGCCcATsT 
                 21% 
                 0% 
                 36% 
                 6% 
               
               
                   
               
               
                 ND8463 
                 1445 
                 GcAAcuucAucuucGccuGTsT 
                 1446 
                 cAGGCGAAGAUGAAGUUGCTsT 
                 80% 
                 4% 
                 84% 
                 13%  
               
               
                   
               
               
                 ND8464 
                 1447 
                 cAAcuucAucuucGccuGcTsT 
                 1448 
                 GcAGGCGAAGAUGAAGUUGTsT 
                 101%  
                 1% 
                 102%  
                 14%  
               
               
                   
               
               
                 ND8465 
                 1449 
                 AAcuucAucuucGccuGccTsT 
                 1450 
                 GGcAGGCGAAGAUGAAGUUTsT 
                 100%  
                 4% 
                 95% 
                 12%  
               
               
                   
               
               
                 ND8466 
                 1451 
                 AcuucAucuucGccuGccGTsT 
                 1452 
                 CGGcAGGCGAAGAUGAAGUTsT 
                 51% 
                 4% 
                 49% 
                 5% 
               
               
                   
               
               
                 ND8467 
                 1453 
                 cuucAucuucGccuGccGcTsT 
                 1454 
                 GCGGcAGGCGAAGAUGAAGTsT 
                 95% 
                 5% 
                 89% 
                 4% 
               
               
                   
               
               
                 ND8468 
                 1455 
                 ucAucuucGccuGccGcuuTsT 
                 1456 
                 AAGCGGcAGGCGAAGAUGATsT 
                 91% 
                 4% 
                 85% 
                 6% 
               
               
                   
               
               
                 ND8469 
                 1457 
                 cAucuucGccuGccGcuucTsT 
                 1458 
                 GAAGCGGcAGGCGAAGAUGTsT 
                 66% 
                 4% 
                 55% 
                 4% 
               
               
                   
               
               
                 ND8470 
                 1459 
                 ucuucGccuGccGcuucAATsT 
                 1460 
                 UUGAAGCGGcAGGCGAAGATsT 
                 97% 
                 2% 
                 99% 
                 11%  
               
               
                   
               
               
                 ND8471 
                 1461 
                 cGccuGccGcuucAAccAGTsT 
                 1462 
                 CUGGUUGAAGCGGcAGGCGTsT 
                 96% 
                 4% 
                 100%  
                 7% 
               
               
                   
               
               
                 ND8472 
                 1463 
                 GccuGccGcuucAAccAGGTsT 
                 1464 
                 CCUGGUUGAAGCGGcAGGCTsT 
                 90% 
                 4% 
                 82% 
                 5% 
               
               
                   
               
               
                 ND8473 
                 1465 
                 AuuAcucucAcuuccAccATsT 
                 1466 
                 UGGUGGAAGUGAGAGuAAUTsT 
                 81% 
                 3% 
                 72% 
                 4% 
               
               
                   
               
               
                 ND8474 
                 1467 
                 uuAcucucAcuuccAccAcTsT 
                 1468 
                 GUGGUGGAAGUGAGAGuAATsT 
                 72% 
                 2% 
                 76% 
                 9% 
               
               
                   
               
               
                 ND8475 
                 1469 
                 AcucucAcuuccAccAcccTsT 
                 1470 
                 GGGUGGUGGAAGUGAGAGUTsT 
                 90% 
                 3% 
                 97% 
                 4% 
               
               
                   
               
               
                 ND8476 
                 1471 
                 ucuGcAcccucAAucccuATsT 
                 1472 
                 uAGGGAUUGAGGGUGcAGATsT 
                 61% 
                 1% 
                 63% 
                 3% 
               
               
                   
               
               
                 ND8477 
                 1473 
                 cuGcAcccucAAucccuAcTsT 
                 1474 
                 GuAGGGAUUGAGGGUGcAGTsT 
                 74% 
                 3% 
                 73% 
                 1% 
               
               
                   
               
               
                 ND8478 
                 1475 
                 uGcAcccucAAucccuAcATsT 
                 1476 
                 UGuAGGGAUUGAGGGUGcATsT 
                 98% 
                 4% 
                 85% 
                 1% 
               
               
                   
               
               
                 ND8479 
                 1477 
                 AcccucAAucccuAcAGGuTsT 
                 1478 
                 ACCUGuAGGGAUUGAGGGUTsT 
                 55% 
                 5% 
                 48% 
                 3% 
               
               
                   
               
               
                 ND8480 
                 1479 
                 cccucAAucccuAcAGGuATsT 
                 1480 
                 uACCUGuAGGGAUUGAGGGTsT 
                 20% 
                 1% 
                 14% 
                 1% 
               
               
                   
               
               
                 ND8481 
                 1481 
                 ccucAAucccuAcAGGuAcTsT 
                 1482 
                 GuACCUGuAGGGAUUGAGGTsT 
                 40% 
                 2% 
                 31% 
                 3% 
               
               
                   
               
               
                 ND8482 
                 1483 
                 AAccAGAAcAAAucGGAcuTsT 
                 1484 
                 AGUCCGAUUUGUUCUGGUUTsT 
                 57% 
                 2% 
                 52% 
                 0% 
               
               
                   
               
               
                 ND8483 
                 1485 
                 AAcAAAucGGAcuGcuucuTsT 
                 1486 
                 AGAAGcAGUCCGAUUUGUUTsT 
                 102%  
                 5% 
                 86% 
                 12%  
               
               
                   
               
               
                 ND8484 
                 1487 
                 AcAAAucGGAcuGcuucuATsT 
                 1488 
                 uAGAAGcAGUCCGAUUUGUTsT 
                 40% 
                 2% 
                 28% 
                 3% 
               
               
                   
               
               
                 ND8485 
                 1489 
                 cAAAucGGAcuGcuucuAcTsT 
                 1490 
                 GuAGAAGcAGUCCGAUUUGTsT 
                 41% 
                 4% 
                 38% 
                 2% 
               
               
                   
               
               
                 ND8486 
                 1491 
                 GcAcccucAAucccuAcAGTsT 
                 1492 
                 CUGuAGGGAUUGAGGGUGCTsT 
                 91% 
                 7% 
                 94% 
                 4% 
               
               
                   
               
               
                 ND8487 
                 1493 
                 ccucAAcAucAAccucAAcTsT 
                 1494 
                 GUUGAGGUUGAUGUUGAGGTsT 
                 46% 
                 2% 
                 37% 
                 3% 
               
               
                   
               
               
                 ND8488 
                 1495 
                 cucAAcAucAAccucAAcuTsT 
                 1496 
                 AGUUGAGGUUGAUGUUGAGTsT 
                 48% 
                 2% 
                 39% 
                 3% 
               
               
                   
               
               
                 ND8489 
                 1497 
                 ucAAcAucAAccucAAcucTsT 
                 1498 
                 GAGUUGAGGUUGAUGUUGATsT 
                 17% 
                 1% 
                 17% 
                 1% 
               
               
                   
               
               
                 ND8490 
                 1499 
                 uAccGcuuccAcuAcAucATsT 
                 1500 
                 UGAUGuAGUGGAAGCGGuATsT 
                 90% 
                 5% 
                 74% 
                 8% 
               
               
                   
               
               
                 ND8491 
                 1501 
                 AccGcuuCCACuAcAucAATsT 
                 1502 
                 UUGAUGuAGUGGAAGCGGUTsT 
                 103%  
                 5% 
                 91% 
                 15%  
               
               
                   
               
               
                 ND8492 
                 1503 
                 GcuuccAcuAcAucAAcAuTsT 
                 1504 
                 AUGUUGAUGuAGUGGAAGCTsT 
                 85% 
                 5% 
                 71% 
                 10%  
               
               
                   
               
               
                 ND8493 
                 1505 
                 cuuccAcuAcAucAAcAucTsT 
                 1506 
                 GAUGUUGAUGuAGUGGAAGTsT 
                 60% 
                 5% 
                 45% 
                 3% 
               
               
                   
               
               
                 ND8494 
                 1507 
                 uccAcuAcAucAAcAuccuTsT 
                 1508 
                 AGGAUGUUGAUGuAGUGGATsT 
                 33% 
                 3% 
                 41% 
                 3% 
               
               
                   
               
               
                 ND8495 
                 1509 
                 ccAcuAcAucAAcAuccuGTsT 
                 1510 
                 CAGGAUGUUGAUGUAGUGGTsT 
                 60% 
                 5% 
                 55% 
                 2% 
               
               
                   
               
               
                 ND8496 
                 1511 
                 cuGGGcAAcuucAucuucGTsT 
                 1512 
                 CGAAGAUGAAGUUGCCcAGTsT 
                 18% 
                 0% 
                 20% 
                 0% 
               
               
                   
               
               
                 ND8497 
                 1513 
                 GGcAAcuucAucuucGccuTST 
                 1514 
                 AGGCGAAGAUGAAGUUGCCTsT 
                 76% 
                 1% 
                 77% 
                 2% 
               
               
                   
               
               
                 ND8498 
                 1515 
                 uucAucuucGccuGccGcuTsT 
                 1516 
                 AGCGGcAGGCGAAGAUGAATsT 
                 65% 
                 4% 
                 74% 
                 12%  
               
               
                   
               
               
                 ND8499 
                 1517 
                 ucGccuGccGcuucAAccATsT 
                 1518 
                 UGGUUGAAGCGGcAGGCGATsT 
                 86% 
                 5% 
                 77% 
                 3% 
               
               
                   
               
               
                 ND-8653 
                 1519 
                 AAUCGGACUGCUUCUACCATsT 
                 1520 
                 UGGUAGAAGCAGUCCGAUUTsT 
                 16% 
                 2% 
                 20% 
                 3% 
               
               
                   
               
               
                 ND-8654 
                 1521 
                 AUCGGACUGCUUCUACCAGTsT 
                 1522 
                 CUGGUAGAAGCAGUCCGAUTsT 
                 54% 
                 8% 
                 67% 
                 11%  
               
               
                   
               
               
                 ND-8655 
                 1523 
                 AAAUCGGACUGCUUCUACCTsT 
                 1524 
                 GGUAGAAGCAGUCCGAUUUTsT 
                 25% 
                 4% 
                 28% 
                 2% 
               
               
                   
               
               
                 ND-8656 
                 1525 
                 UCGGACUGCUUCUACCAGATsT 
                 1526 
                 UCUGGUAGAAGCAGUCCGATsT 
                 12% 
                 2% 
                 17% 
                 1% 
               
               
                   
               
               
                 ND-8657 
                 1527 
                 ACCAGAACAAAUCGGACUGTsT 
                 1528 
                 CAGUCCGAUUUGUUCUGGUTsT 
                 33% 
                 3% 
                 35% 
                 1% 
               
               
                   
               
               
                 ND-8658 
                 1529 
                 CCAGAACAAAUCGGACUGCTsT 
                 1530 
                 GCAGUCCGAUUUGUUCUGGTsT 
                 27% 
                 3% 
                 30% 
                 2% 
               
               
                   
               
               
                 ND-8659 
                 1531 
                 CAGAACAAAUCGGACUGCUTsT 
                 1532 
                 AGCAGUCCGAUUUGUUCUGTsT 
                 15% 
                 1% 
                 22% 
                 3% 
               
               
                   
               
               
                 ND-8660 
                 1533 
                 CUUCGCCUGCCGCUUCAACTsT 
                 1534 
                 GUUGAAGCGGCAGGCGAAGTsT 
                 69% 
                 17%  
                 75% 
                 10%  
               
               
                   
               
               
                 ND-8661 
                 1535 
                 UGGUACCGCUUCCACUACATsT 
                 1536 
                 UGUAGUGGAAGCGGUACCATsT 
                 16% 
                 2% 
                 20% 
                 3% 
               
               
                   
               
               
                 ND-8662 
                 1537 
                 AUCUUCGCCUGCCGCUUCATsT 
                 1538 
                 UGAAGCGGCAGGCGAAGAUTsT 
                 19% 
                 2% 
                 25% 
                 4% 
               
               
                   
               
               
                 ND-8663 
                 1539 
                 UUCGCCUGCCGCUUCAACCTsT 
                 1540 
                 GGUUGAAGCGGCAGGCGAATsT 
                 90% 
                 4% 
                 97% 
                 10%  
               
               
                   
               
               
                 ND-8664 
                 1541 
                 CACCCUCAAUCCCUACAGGTsT 
                 1542 
                 CCUGUAGGGAUUGAGGGUGTsT 
                 19% 
                 2% 
                 25% 
                 3% 
               
               
                   
               
               
                 ND-8665 
                 1543 
                 AGAACAAAUCGGACUGCUUTsT 
                 1544 
                 AAGCAGUCCGAUUUGUUCUTsT 
                 13% 
                 1% 
                 22% 
                 2% 
               
               
                   
               
               
                 ND-8666 
                 1545 
                 GAACAAAUCGGACUGCUUCTsT 
                 1546 
                 GAAGCAGUCCGAUUUGUUCTsT 
                 11% 
                 2% 
                 18% 
                 2% 
               
               
                   
               
               
                 ND-8667 
                 1547 
                 CGGACUGCUUCUACCAGACTsT 
                 1548 
                 GUCUGGUAGAAGCAGUCCGTsT 
                 13% 
                 1% 
                 16% 
                 2% 
               
               
                   
               
               
                 ND-8668 
                 1549 
                 AGCCUCAACAUCAACCUCATsT 
                 1550 
                 UGAGGUUGAUGUUGAGGCUTsT 
                 17% 
                 4% 
                 21% 
                 3% 
               
               
                   
               
               
                 ND-8669 
                 1551 
                 CGGUCAACAUCAACCUCAATsT 
                 1552 
                 UUGAGGUUGAUGUUGAGGCTsT 
                 13% 
                 1% 
                 21% 
                 3% 
               
               
                   
               
               
                 ND-8670 
                 1553 
                 GUCAGCCUCAACAUCAACCTsT 
                 1554 
                 GGUUGAUGUUGAGGCUGACTsT 
                 43% 
                 11%  
                 27% 
                 3% 
               
               
                   
               
               
                 ND-8671 
                 1555 
                 UCAGCCUCAACAUCAACCUTsT 
                 1556 
                 AGGUUGAUGUUGAGGCUGATsT 
                 90% 
                 17%  
                 53% 
                 13%  
               
               
                   
               
               
                 ND-8672 
                 1557 
                 CAGCCUCAACAUCAACCUCTsT 
                 1558 
                 GAGGUUGAUGUUGAGGCUGTsT 
                 17% 
                 3% 
                 11% 
                 3% 
               
               
                   
               
               
                 ND-8673 
                 1559 
                 GGAGCUGGACCGCAUCACATsT 
                 1560 
                 UGUGAUGCGGUCCAGCUCCTsT 
                 25% 
                 3% 
                 18% 
                 3% 
               
               
                   
               
               
                 ND-8674 
                 1561 
                 GUACCGCUUCCACUACAUCTsT 
                 1562 
                 GAUGUAGUGGAAGCGGUACTsT 
                 21% 
                 4% 
                 16% 
                 4% 
               
               
                   
               
               
                 ND-8675 
                 1563 
                 CCGCUUCCACUACAUCAACTsT 
                 1564 
                 GUUGAUGUAGUGGAAGCGGTsT 
                 25% 
                 4% 
                 19% 
                 3% 
               
               
                   
               
               
                 ND-8676 
                 1565 
                 CGCUUCCACUACAUCAACATsT 
                 1566 
                 UGUUGAUGUAGUGGAAGCGTsT 
                 16% 
                 3% 
                 14% 
                 1% 
               
               
                   
               
               
                 ND-8677 
                 1567 
                 UUCCACUACAUCAACAUCCTsT 
                 1568 
                 GGAUGUUGAUGUAGUGGAATsT 
                 110%  
                 19%  
                 97% 
                 9% 
               
               
                   
               
               
                 ND-8678 
                 1569 
                 UGGGCAACUUCAUCUUCGCTsT 
                 1570 
                 GCGAAGAUGAAGUUGCCCATsT 
                 50% 
                 8% 
                 40% 
                 5% 
               
               
                   
               
               
                 ND-8679 
                 1571 
                 GCAACUUCAUCUUCGCCUGTsT 
                 1572 
                 CAGGCGAAGAUGAAGUUGCTsT 
                 19% 
                 3% 
                 17% 
                 2% 
               
               
                   
               
               
                 ND-8680 
                 1573 
                 CAACUUCAUCUUCGCCUGCTsT 
                 1574 
                 GCAGGCGAAGAUGAAGUUGTsT 
                 25% 
                 2% 
                 23% 
                 2% 
               
               
                   
               
               
                 ND-8681 
                 1575 
                 AACUUCAUCUUCGCCUGCCTsT 
                 1576 
                 GGCAGGCGAAGAUGAAGUUTsT 
                 104%  
                 7% 
                 85% 
                 10%  
               
               
                   
               
               
                 ND-8682 
                 1577 
                 ACUUCAUCUUCGCCUGCCGTsT 
                 1578 
                 CGGCAGGCGAAGAUGAAGUTsT 
                 91% 
                 8% 
                 63% 
                 9% 
               
               
                   
               
               
                 ND-8683 
                 1579 
                 CUUCAUCUUCGCCUGCCGCTsT 
                 1580 
                 GCGGCAGGCGAAGAUGAAGTsT 
                 88% 
                 6% 
                 58% 
                 6% 
               
               
                   
               
               
                 ND-8684 
                 1581 
                 UCAUCUUCGCCUGCCGCUUTsT 
                 1582 
                 AAGCGGCAGGCGAAGAUGATsT 
                 76% 
                 3% 
                 64% 
                 4% 
               
               
                   
               
               
                 ND-8685 
                 1583 
                 CAUCUUCGCCUGCCGCUUCTsT 
                 1584 
                 GAAGCGGCAGGCGAAGAUGTsT 
                 15% 
                 1% 
                 18% 
                 3% 
               
               
                   
               
               
                 ND-8686 
                 1585 
                 UCUUCGCCUGCCGCUUCAATsT 
                 1586 
                 UUGAAGCGGCAGGCGAAGATsT 
                 109%  
                 22%  
                 31% 
                 3% 
               
               
                   
               
               
                 ND-8687 
                 1587 
                 CGCCUGCCGCUUCAACCAGTsT 
                 1588 
                 CUGGUUGAAGCGGCAGGCGTsT 
                 90% 
                 21%  
                 49% 
                 2% 
               
               
                   
               
               
                 ND-8688 
                 1589 
                 GCCUGCCGCUUCAACCAGGTsT 
                 1590 
                 CCUGGUUGAAGCGGCAGGCTsT 
                 43% 
                 9% 
                 24% 
                 7% 
               
               
                   
               
               
                 ND-8689 
                 1591 
                 AUUACUCUCACUUCCACCATsT 
                 1592 
                 UGGUGGAAGUGAGAGUAAUTsT 
                 27% 
                 4% 
                 19% 
                 2% 
               
               
                   
               
               
                 ND-8690 
                 1593 
                 UUACUCUCACUUCCACCACTsT 
                 1594 
                 GUGGUGGAAGUGAGAGUAATsT 
                 109%  
                 7% 
                 85% 
                 8% 
               
               
                   
               
               
                 ND-8691 
                 1595 
                 ACUCUCACUUCCACCACCCTsT 
                 1596 
                 GGGUGGUGGAAGUGAGAGUTsT 
                 93% 
                 11%  
                 87% 
                 12%  
               
               
                   
               
               
                 ND-8692 
                 1597 
                 UCUGCACCCUCAAUCCCUATsT 
                 1598 
                 UAGGGAUUGAGGGUGCAGATsT 
                 31% 
                 12%  
                 17% 
                 2% 
               
               
                   
               
               
                 ND-8693 
                 1599 
                 CUGCACCCUCAAUCCCUACTsT 
                 1600 
                 GUAGGGAUUGAGGGUGCAGTsT 
                 41% 
                 25%  
                 31% 
                 4% 
               
               
                   
               
               
                 ND-8694 
                 1601 
                 UGCACCCUCAAUCCCUACATsT 
                 1602 
                 UGUAGGGAUUGAGGGUGCATsT 
                 75% 
                 25%  
                 43% 
                 3% 
               
               
                   
               
               
                 ND-8695 
                 1603 
                 ACCCUCAAUCCCUACAGGUTsT 
                 1604 
                 ACCUGUAGGGAUUGAGGGUTsT 
                 65% 
                 26%  
                 25% 
                 5% 
               
               
                   
               
               
                 ND-8696 
                 1605 
                 CCCUCAAUCCCUACAGGUATsT 
                 1606 
                 UACCUGUAGGGAUUGAGGGTsT 
                 18% 
                 2% 
                 13% 
                 1% 
               
               
                   
               
               
                 ND-8697 
                 1607 
                 CCUCAAUCCCUACAGGUACTsT 
                 1608 
                 GUACCUGUAGGGAUUGAGGTsT 
                 16% 
                 4% 
                 13% 
                 2% 
               
               
                   
               
               
                 ND-8698 
                 1609 
                 AACCAGAACAAAUCGGACUTsT 
                 1610 
                 AGUCCGAUUUGUUCUGGUUTsT 
                 40% 
                 2% 
                 30% 
                 2% 
               
               
                   
               
               
                 ND-8699 
                 1611 
                 AACAAAUCGGACUGCUUCUTsT 
                 1612 
                 AGAAGCAGUCCGAUUUGUUTsT 
                 56% 
                 4% 
                 45% 
                 3% 
               
               
                   
               
               
                 ND-8700 
                 1613 
                 ACAAAUCGGACUGCUUCUATsT 
                 1614 
                 UAGAAGCAGUCCGAUUUGUTsT 
                 18% 
                 3% 
                 12% 
                 1% 
               
               
                   
               
               
                 ND-8701 
                 1615 
                 CAAAUCGGACUGCUUCUACTsT 
                 1616 
                 GUAGAAGCAGUCCGAUUUGTsT 
                 15% 
                 2% 
                 15% 
                 4% 
               
               
                   
               
               
                 ND-8702 
                 1617 
                 GCACCCUCAAUCCCUACAGTsT 
                 1618 
                 CUGUAGGGAUUGAGGGUGCTsT 
                 53% 
                 4% 
                 46% 
                 20%  
               
               
                   
               
               
                 ND-8703 
                 1619 
                 CCUCAACAUCAACCUCAACTsT 
                 1620 
                 GUUGAGGUUGAUGUUGAGGTsT 
                 25% 
                 6% 
                 26% 
                 9% 
               
               
                   
               
               
                 ND-8704 
                 1621 
                 CUCAACAUCAACCUCAACUTsT 
                 1622 
                 AGUUGAGGUUGAUGUUGAGTsT 
                 30% 
                 8% 
                 37% 
                 26%  
               
               
                   
               
               
                 ND-8705 
                 1623 
                 UCAACAUCAACCUCAACUCTsT 
                 1624 
                 GAGUUGAGGUUGAUGUUGATsT 
                 55% 
                 1% 
                 50% 
                 10%  
               
               
                   
               
               
                 ND-8706 
                 1625 
                 UACCGCUUCCACUACAUCATsT 
                 1626 
                 UGAUGUAGUGGAAGCGGUATsT 
                 36% 
                 7% 
                 31% 
                 7% 
               
               
                   
               
               
                 ND-8707 
                 1627 
                 ACCGCUUCCACUACAUCAATsT 
                 1628 
                 UUGAUGUAGUGGAAGCGGUTsT 
                 23% 
                 5% 
                 27% 
                 10%  
               
               
                   
               
               
                 ND-8708 
                 1629 
                 GCUUCCACUACAUCAACAUTsT 
                 1630 
                 AUGUUGAUGUAGUGGAAGCTsT 
                 16% 
                 4% 
                 24% 
                 12%  
               
               
                   
               
               
                 ND-8709 
                 1631 
                 CUUCCACUACAUCAACAUCTsT 
                 1632 
                 GAUGUUGAUGUAGUGGAAGTsT 
                 62% 
                 3% 
                 74% 
                 27%  
               
               
                   
               
               
                 ND-8710 
                 1633 
                 UCCACUACAUCAACAUCCUTsT 
                 1634 
                 AGGAUGUUGAUGUAGUGGATsT 
                 45% 
                 8% 
                 41% 
                 1% 
               
               
                   
               
               
                 ND-8711 
                 1635 
                 CCACUACAUCAACAUCCUGTsT 
                 1636 
                 CAGGAUGUUGAUGUAGUGGTsT 
                 23% 
                 4% 
                 27% 
                 10%  
               
               
                   
               
               
                 ND-8712 
                 1637 
                 CUGGGCAACUUCAUCUUCGTsT 
                 1638 
                 CGAAGAUGAAGUUGCCCAGTsT 
                 34% 
                 4% 
                 26% 
                 5% 
               
               
                   
               
               
                 ND-8713 
                 1639 
                 GGCAACUUCAUCUUCGCCUTsT 
                 1640 
                 AGGCGAAGAUGAAGUUGCCTsT 
                 30% 
                 3% 
                 23% 
                 2% 
               
               
                   
               
               
                 ND-8714 
                 1641 
                 UUCAUCUUCGCCUGCCGCUTsT 
                 1642 
                 AGCGGCAGGCGAAGAUGAATsT 
                 90% 
                 14%  
                 85% 
                 14%  
               
               
                   
               
               
                 ND-8715 
                 1643 
                 UCGCCUGCCGCUUCAACCATsT 
                 1644 
                 UGGUUGAAGCGGCAGGCGATsT 
                 23% 
                 2% 
                 20% 
                 4% 
               
               
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2A 
               
             
            
               
                   
               
               
                 Concentration at 50% inhibition (IC50) 
               
               
                 for exemplary iRNA agents of Table 1A 
               
            
           
           
               
               
               
               
            
               
                   
                   
                 IC50 [nM] 
                 IC50 [nM] 
               
               
                   
                   
                 1st DRC in 
                 2nd DRC in 
               
               
                   
                 Duplex ID 
                 H441 
                 H441 
               
               
                   
                   
               
            
           
           
               
               
               
               
            
               
                   
                 ND8294 
                 0.1949 
                 0.0468 
               
               
                   
                 ND8295 
                 0.1011 
                 0.0458 
               
               
                   
                 ND8299 
                 0.5986 
                 0.5638 
               
               
                   
                 ND8302 
                 0.0144 
                 0.0134 
               
               
                   
                 ND8313 
                 0.0315 
                 0.0124 
               
               
                   
                 ND8320 
                 0.0796 
                 0.0078 
               
               
                   
                 ND8331 
                 0.0213 
                 0.0158 
               
               
                   
                 ND8332 
                 0.0205 
                 0.0089 
               
               
                   
                 ND8343 
                 0.0523 
                 0.0293 
               
               
                   
                 ND8348 
                 0.0156 
                 0.0182 
               
               
                   
                 ND8356 
                 0.0241 
                 0.0099 
               
               
                   
                 ND8357 
                 0.0054 
                 0.0032 
               
               
                   
                 ND8363 
                 0.1186 
                 0.0337 
               
               
                   
                 ND8368 
                 0.0487 
                 0.1209 
               
               
                   
                 ND8371 
                 0.0811 
                 0.0911 
               
               
                   
                 ND8372 
                 0.0584 
                 0.0425 
               
               
                   
                 ND8373 
                 0.0066 
                 0.0165 
               
               
                   
                 ND8375 
                 0.1176 
                 0.1187 
               
               
                   
                 ND8380 
                 0.6817 
                 0.5747 
               
               
                   
                 ND8381 
                 0.0037 
                 0.0041 
               
               
                   
                 ND8383 
                 0.0275 
                 0.1257 
               
               
                   
                 ND8384 
                 0.0357 
                 0.0082 
               
               
                   
                 ND8391 
                 0.0260 
                 0.0349 
               
               
                   
                 ND8392 
                 0.3831 
                 0.4775 
               
               
                   
                 ND8396 
                 0.0023 
                 0.0052 
               
               
                   
                 ND8403 
                 0.0808 
                 0.0759 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2B 
               
             
            
               
                   
               
               
                 Concentration at 50% inhibition (IC50) and 
               
               
                 for exemplary iRNA agents of Table 1D. 
               
            
           
           
               
               
               
               
            
               
                   
                   
                 IC50 [nM] 
                 IC50 [nM] 
               
               
                   
                   
                 1st DRC in 
                 2nd DRC in 
               
               
                   
                 Duplex ID 
                 H441 
                 H441 
               
               
                   
                   
               
            
           
           
               
               
               
               
            
               
                   
                 ND8441 
                 0.6738 
                 0.8080 
               
               
                   
                 ND8443 
                 0.0346 
                 0.0263 
               
               
                   
                 ND8449 
                 0.0120 
                 0.0067 
               
               
                   
                 ND8450 
                 0.0257 
                 0.0106 
               
               
                   
                 ND8451 
                 0.1320 
                 0.0931 
               
               
                   
                 ND8453 
                 0.0079 
                 0.0033 
               
               
                   
                 ND8489 
                 0.1640 
                 0.1593 
               
               
                   
                 ND8496 
                 0.0387 
                 0.0185 
               
               
                   
                   
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2C 
               
             
            
               
                   
               
               
                 % Activity of the exemplary RNAi towards inhibition of alpha- 
               
               
                 ENaC gene expression in the assays described in Example 3 
               
            
           
           
               
               
               
               
            
               
                   
                   
                 % alpha-ENaC 
                 Cynomolgous 
               
               
                   
                   
                 expression in 
                 alpha-ENaC 
               
               
                   
                   
                 primary HBEC 
                 expression 
               
               
                   
                   
                 (% of control) 
                 (% of control) 
               
               
                   
                 Duplex identifier 
                 50 nM siRNA 
                 45 nM siRNA 
               
               
                   
                   
               
            
           
           
               
               
               
               
            
               
                   
                 Untransfected 
                 77.2 
                 n/a 
               
               
                   
                 Non-targetting Control 
                 100 
                 93.3 
               
               
                   
                 Negative Control 
                 n/a 
                 100 
               
               
                   
                 (Non-cyno alpha-ENaC) 
               
               
                   
                 ND8449 
               
               
                   
                 ND-8302 
                 30.2 
                 57 
               
               
                   
                 ND-8332 
                 24.7 
                 54.3 
               
               
                   
                 ND-8348 
                 40.1 
                 56.2 
               
               
                   
                 ND-8356 
                 36.6 
                 55.8 
               
               
                   
                 ND-8357 
                 29.6 
                 50.4 
               
               
                   
                 ND-8373 
                 30.4 
                 53.8 
               
               
                   
                 ND-8381 
                 32.5 
                 40.4 
               
               
                   
                 ND-8396 
                 34.1 
                 46.3 
               
               
                   
                 ND-8450 
                 45.9 
                 78.9 
               
               
                   
                 ND-8453 
                 30.1 
                 55.3