Exon 17-targeted nucleic acids, compositions, and methods for treatment of dystrophin-based myopathies

Abstract

The disclosure relates to the field of gene therapy for the treatment of a muscular dystrophy including, but not limited to, Duchenne Muscular Dystrophy (DMD). More particularly, the disclosure provides nucleic acids, including nucleic acids encoding U7-based small nuclear ribonucleic acids (RNAs) (snRNAs), U7-based snRNAs, and vectors (including, but not limited to, recombinant adeno-associated virus (rAAV)), nanoparticles, extracellular vesicles, or exosomes comprising the nucleic acids to induce exon-skipping for use in treating a muscular dystrophy including, but not limited to, DMD, resulting from a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17) including, but not limited to, any mutation involving, surrounding, or affecting DMD exon 17.

Description

EXON 17-TARGETED NUCLEIC ACIDS, COMPOSITIONS, AND METHODS FOR TREATMENT OF DYSTROPHIN-BASED MYOPATHIESSTATEMENT OF GOVERNMENT INTEREST

[0001] This invention was made with government support under AR070604 awarded by the National Institutes of Health. The government has certain rights in the invention.FIELD

[0002] The disclosure relates to the field of gene therapy for the treatment of muscular dystrophy. More particularly, the disclosure provides nucleic acids, including nucleic acids encoding U7-based small nuclear ribonucleic acids (RNAs) (snRNAs), U7-based snRNAs, and vectors (including, but not limited to, recombinant adeno-associated virus (rAAV)), nanoparticles, extracellular vesicles, or exosomes comprising the nucleic acids to deliver such nucleic acids, including, in some aspects, nucleic acids encoding U7-based snRNAs, to induce exon-skipping in a cell or for use in treating, preventing, or ameliorating a muscular dystrophy resulting from a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17) including, but not limited to, any mutation involving, surrounding, or affecting DMD exon 17.INCORPORATION BY REFERENCE OF THE SEQUENCE LISTING

[0003] This application contains, as a separate part of disclosure, a Sequence Listing in computer-readable form (Filename: 56872_SeqListing.xml; Size: 564,185 bytes; Created: February 21 , 2025) which is incorporated by reference herein in its entirety.BACKGROUND

[0004] Muscular dystrophies (MDs) are a group of genetic degenerative diseases primarily affecting voluntary muscles. The group is characterized by progressive weakness and degeneration of the skeletal muscles that control movement. Some forms of MD develop in infancy or childhood, while others may not appear until middle age or later. The disorders differ in terms of the distribution and extent of muscle weakness (some forms of MD also affect cardiac muscle), the age of onset, the rate of progression, and the pattern of inheritance.

[0005] The MDs are a group of diseases without identifiable treatment that gravely impact individuals, families, and communities. The costs are incalculable. Individuals suffer emotional strain and reduced quality of life associated with loss of self-esteem. Extreme physical challenges resulting from loss of limb function creates hardships in activities of daily living. Family dynamics suffer through financial loss and challenges to interpersonal relationships.Siblings of the affected feel estranged, and strife between spouses often leads to divorce, especially if responsibility for the muscular dystrophy can be laid at the feet of one of the parental partners. The burden of quest to find a cure often becomes a life-long, highly focused effort that detracts and challenges every aspect of life. Beyond the family, the community bears a financial burden through the need for added facilities to accommodate the handicaps of the muscular dystrophy population in special education, special transportation, and costs for recurrent hospitalizations to treat recurrent respiratory tract infections and cardiac complications. Financial responsibilities are shared by state and federal governmental agencies extending the responsibilities to the taxpaying community.

[0006] One form of MD is Duchenne Muscular Dystrophy (DMD). DMD is one of the severe forms of muscular dystrophy, affecting approximately one in every 3,600-5,200 newborn males and characterized by progressive muscle wasting and weakness, cardiovascular, and respiratory complications. These lead to loss of ambulation by age 12 and premature death by the 3rd or 4th decade of life. DMD is caused by mutations in the dystrophin gene leading to absence of dystrophin protein (427 KDa) in skeletal and cardiac muscles, as well as the gastrointestinal tract and retina. Dystrophin not only protects the sarcolemma from eccentric contractions, but also anchors a number of signaling proteins in close proximity to sarcolemma. Another form of MD is Becker Muscular Dystrophy (BMD). BMD, like DMD, is a genetic disorder that gradually makes the body's muscles weaker and smaller. BMD affects the muscles of the hips, pelvis, thighs, and shoulders, as well as the heart, but is known to cause less severe problems than DMD.

[0007] Many clinical cases of DMD are linked to deletion mutations in the DMD gene. In contrast to the deletion mutations, DMD exon duplications account for around 5% of diseasecausing mutations in unbiased samples of dystrophinopathy patients [Dent et al., Am J Med Genet, 134(3): 295-298 (2005)], although in some catalogues of mutations the number of duplications is higher, including that published by the United Dystrophinopathy Project by Flanigan et al. [Hum Mutat, 30(12): 1657-1666 (2009)], in which it was 1 1%. BMD is also caused by a change in the dystrophin gene, which makes the protein too short. The flawed dystrophin puts muscle cells at risk for damage with normal use. See also, U.S. Patent Application Publication Nos. 2012 / 0077860, published March 29, 2012; 2013 / 0072541 , published March 21 , 2013; and 2013 / 0045538, published February 21 , 2013.

[0008] Despite many lines of research following the identification of the dystrophin gene, treatment options are limited. There thus remains a need in the art for treatments for MDs,including DMD. The most advanced therapies include those that aim at restoration of the missing protein, dystrophin, using mutation-specific genetic approaches, such as antisense oligonucleotide (AON)-mediated exon skipping. The disclosure provides an approach to treating patients carrying exon 17 skip-amenable mutations including, but not limited to, mutations that are within or that flank exon 17 of the DMD gene. Such mutations are known to affect up to 5.5% of all DMD patients.SUMMARY

[0009] The disclosure provides products, methods, and uses for a new gene therapy for treating, ameliorating, delaying the progression of, and / or preventing a muscular dystrophy involving a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17) including, but not limited to, any mutation involving, surrounding, or affecting DMD exon 17. More particularly, the disclosure provides nucleic acids, U7-based small nuclear ribonucleic acids (RNAs) (snRNAs), and compositions, vectors, nanoparticles, extracellular vesicles, or exosomes comprising the nucleic acids to induce exon-skipping to provide an altered form of dystrophin protein for use in treating a muscular dystrophy resulting from any mutation involving, surrounding, or affecting DMD exon 17.

[0010] The disclosure provides a nucleic acid that binds or is complementary to a polynucleotide encoding(a) exon 17 of the DMD gene comprising intronic sequence surrounding exon 17 comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 or 2;(b) exon 17 of the DMD gene or its reverse complementary sequence comprising or consisting of SEQ ID NO: 3 or 4; or(c) the amino acid sequence of SEQ ID NO: 5.

[0011] The disclosure provides a nucleic acid comprising:(a) an antisense encoding nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 , or comprising or consisting of the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 ;(b) an antisense encoding nucleotide sequence that binds to a target nucleotide sequence of SEQ ID NO: 6, 12, 18, 24, or 30;(c) an antisense encoding nucleotide sequence that binds to a target mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32; or(d) an antisense encoding nucleotide sequence that binds to a target antisense mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 9, 15, 21 , 27, or 33, or comprising or consisting of the nucleotide sequence of 9, 15, 21 , 27, or 33.

[0012] In some aspects, the nucleic acid further comprises a promoter and / or enhancer. In some aspects a plasmid of the disclosure comprises multiple promoters and / or enhancers. In some aspects, the promoter is a U6, LI7, tRNA, H1 , CMV, minimal CMV, T7, EF1 -alpha, Minimal EF1 -alpha, or a tissue-specific promoter including, but not limited to, a muscle-specific promoter or a cardiac-specific promoter. In some aspects, the promoter is U6 or H1 . In some aspects, the muscle-specific promoter is unc45b, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, alpha-myosin heavy chain enhancer- / MCK enhancer-promoter (MHCK7), or CK1 . In some aspects, the cardiac-specific promoter is alpha-myosin heavy chain enhancer- / MCK enhancer-promoter (MHCK7), the 250-bp fragment of the myosin light chain-2v (MLC-2v) gene promoter (MLC250), cardiac troponin T (cTnT) promoter, the cc-myosin heavy chain (oc-MHC) promoter, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, or CK1 . In some aspects, the promoter is a U7 promoter. In some aspects, the enhancer is the alpha-myosin heavy chain (ocMHC) enhancer (aMHCEnh) or CMV enhancer.

[0013] In some aspects, therefore, a nucleic acid of the disclosure comprises or consists of(a) a nucleotide sequence encoding a U7snRA comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34; or(b) a nucleotide sequence encoding a U7snRNA reverse complement sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 1 1 , 17, 23, 29, or 35, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 11 , 17, 23, 29, or 35.

[0014] In some aspects, a nucleic acid of the disclosure further an inverted terminal repeat region when the nucleic acid is to be delivered via a vector, such as an AAV vector.

[0015] The disclosure also provides a nucleic acid comprising a combination of any two or more of any of the nucleic acids described herein. In some aspects, the combination comprises three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or even ten or more nucleic acids of the disclosure. In some aspects, delivery of the combination increases efficiency of exon skipping.

[0016] The disclosure therefore also provides a nucleic acid wherein the nucleotide sequence comprises or consists of(a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 42-61 , or(b) a nucleotide sequence comprising any one of SEQ ID NOs: 42-61 .

[0017] The disclosure also provides a nucleic acid comprising staffer and / or enhancer sequence in addition to promoter sequence. In some aspects, the inclusion of staffer and / or enhancer sequence increases efficiency of exon skipping. In some aspects, the nucleic acid further comprises a promoter and / or enhancer. In some aspects, the nucleic acid further comprises multiple promoters and / or enhancers. In some aspects, the promoter is a LI7 promoter. In some aspects, the enhancer is the alpha-myosin heavy chain (ocMHC) enhancer (aMHCEnh) or CMV enhancer. In some aspects, the nucleic acid comprises multiple U7 promoters and / or multiple alpha-myosin heavy chain (ocMHC) enhancers (aMHCEnh) or CMV enhancers. In some instances, the nucleic acid further comprises a combination of any two or more of any of the nucleic acids described herein. In some aspects, the combination comprises three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or even ten or more nucleic acids of the disclosure. In some aspects, the presence of these additional elements or additional copies of snRNAs in the construct increases efficiency of exon skipping.

[0018] The disclosure thus also provides a nucleic acid wherein the nucleotide sequence comprises or consists of(a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 62-161 , or(b) a nucleotide sequence comprising any one of SEQ ID NOs: 62-161 .

[0019] The disclosure also provides a composition comprising any one or more of the nucleic acids of the disclosure. In some aspects, the disclosure comprises a composition comprising a vector, nanoparticle, extracellular vesicle, or exosome of the disclosure.

[0020] Thus, the disclosure also provides a vector, nanoparticle, extracellular vesicle, or exosome comprising a nucleic acid of the disclosure. In some aspects, the vector is an adeno- associated virus (AAV) or a recombinant AAV (rAAV). In some aspects, the AAV or rAAV lacks rep and cap genes. In some aspects, the AAV or rAAV is a self-complementary AAV or a self- complementary recombinant AAV (scAAV). In some aspects, the AAV is AAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV.rh74, AAV.rh8, AAV.rhl 0, AAV11 , AAV12, AAV13, AAV-anc80, AAV-B1 , AAV-BR1 , AAV.PHP.EB, AAVv66, AAV2 / 1 , AAV2 / 8, AAV2 / 9, AAVMYO, MYOAAV, MYOAAV1 A, MYOAAV2A, MYOAAV3A, or any other myotropic serotype, or any derivative thereof. In some aspects, the AAV is AAVMYO, MYOAAV, MYOAAV1 A, MYOAAV2A, MYOAAV3A, or AAV9.

[0021] The disclosure provides a method for inducing skipping of exon 17 of the DMD gene in a cell, the method comprising providing the cell with a nucleic acid of the disclosure, a composition of the disclosure, a vector, nanoparticle, extracellular vesicle, or exosome of the disclosure. In some aspects, the vector is an AAV. In some aspects, the nucleic acid is a nucleic acid that binds or is complementary to a polynucleotide encoding (a) exon 17 of the DMD gene comprising intronic sequence surrounding exon 17 comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 or 2; (b) exon 17 of the DMD gene or its reverse complementary sequence comprising or consisting of SEQ ID NO: 3 or 4; or (c) the amino acid sequence of SEQ ID NO: 5. In some aspects, the nucleic acid is a nucleic acid comprising: (a) an antisense encoding nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 , or comprising or consisting of the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 ; (b) an antisense encoding nucleotide sequence that binds to a target nucleotide sequence of SEQ ID NO: 6, 12, 18, 24, or 30; (c) an antisense encoding nucleotide sequence that binds to a target mRNA sequence comprising atleast 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32; or (d) an antisense encoding nucleotide sequence that binds to a target antisense mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 9, 15, 21 , 27, or 33, or comprising or consisting of the nucleotide sequence of 9, 15, 21 , 27, or 33. In some aspects, the nucleic acid further comprises a promoter and / or enhancer. In some aspects, the nucleic acid comprises two or more promoters and / or enhancers. In some aspects, the promoter is a U6, U7, tRNA, H1 , CMV, minimal CMV, T7, EF1 -alpha, Minimal EF1 -alpha, or a tissue-specific promoter including, but not limited to, a muscle-specific promoter or a cardiacspecific promoter. In some aspects, the promoter is U6 or H1 . In some aspects, the musclespecific promoter is unc45b, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, alpha-myosin heavy chain enhancer- / MCK enhancer-promoter (MHCK7), or CK1 . In some aspects, the cardiac-specific promoter is alpha-myosin heavy chain enhancer- / MCK enhancerpromoter (MHCK7), the 250-bp fragment of the myosin light chain-2v (MLC-2v) gene promoter (MLC250), cardiac troponin T (cTnT) promoter, the a-myosin heavy chain (a-MHC) promoter, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, or CK1 . In some aspects, the promoter is a U7 promoter. In some aspects, the enhancer is the alpha-myosin heavy chain (aMHC) enhancer (aMHCEnh) or CMV enhancer. In some aspects, the nucleic acid comprises multiple U7 promoters and / or multiple alpha-myosin heavy chain (aMHC) enhancers (aMHCEnh) or CMV enhancers.

[0022] In some aspects, therefore, the disclosure provides a method for inducing skipping of exon 17 of the DMD gene in a cell, the method comprising providing the cell with a nucleic acid comprising or consisting of (a) a nucleotide sequence encoding a U7snRA comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34; or (b) a nucleotide sequence encoding a U7snRNA reverse complement sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 1 1 , 17, 23, 29, or 35, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 11 , 17, 23, 29, or 35.

[0023] In some aspects, therefore, the disclosure provides a method for inducing skipping of exon 17 of the DMD gene in a cell, the method comprising providing the cell with a nucleic acid comprising or consisting of (a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 42-61 , or (b) a nucleotide sequence comprising any one of SEQ ID NOs: 42-61 .In some aspects, therefore, the disclosure provides a method for inducing skipping of exon 17 of the D / WD gene in a cell, the method comprising providing the cell with a nucleic acid comprising or consisting of (a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 62-161 , or (b) a nucleotide sequence comprising any one of SEQ ID NOs: 62-161.

[0024] In some aspects, the disclosure provides a method for inducing skipping of exon 17 of the DMD gene in a cell, wherein the cell is in a human subject.

[0025] The disclosure also provides a method for treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17) comprising administering to the subject an effective amount of a nucleic acid of the disclosure, a composition of the disclosure, a vector, nanoparticle, extracellular vesicle, or exosome of the disclosure. In some aspects, the vector is an AAV.

[0026] In some aspects, the nucleic acid is a nucleic acid that binds or is complementary to a polynucleotide encoding (a) exon 17 of the DMD gene comprising intronic sequence surrounding exon 17 comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 or 2; (b) exon 17 of the DMD gene or its reverse complementary sequence comprising or consisting of SEQ ID NO: 3 or 4; or (c) the amino acid sequence of SEQ ID NO: 5.

[0027] In some aspects, the nucleic acid is a nucleic acid comprising: (a) an antisense encoding nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 , or comprising or consisting of the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 ; (b) an antisense encoding nucleotide sequence that binds to a target nucleotide sequence of SEQ ID NO: 6, 12, 18, 24, or 30; (c) an antisense encoding nucleotide sequence that binds to a target mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least99% identity to the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32; or (d) an antisense encoding nucleotide sequence that binds to a target antisense mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 9, 15, 21 , 27, or 33, or comprising or consisting of the nucleotide sequence of 9, 15, 21 , 27, or 33.

[0028] In some aspects, the nucleic acid further comprises promoter. In some aspects, the promoter is a U6, U7, tRNA, H1 , CMV, minimal CMV, T7, EF1 -alpha, Minimal EF1 -alpha, or a tissue-specific promoter including, but not limited to, a muscle-specific promoter or a cardiacspecific promoter. In some aspects, the promoter is U6 or H1 . In some aspects, the musclespecific promoter is unc45b, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, alpha-myosin heavy chain enhancer- / MCK enhancer-promoter (MHCK7), or CK1 . In some aspects, the cardiac-specific promoter is alpha-myosin heavy chain enhancer- / MCK enhancerpromoter (MHCK7), the 250-bp fragment of the myosin light chain-2v (MLC-2v) gene promoter (MLC250), cardiac troponin T (cTnT) promoter, the oc-myosin heavy chain (oc-MHC) promoter, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, or CK1 . In some aspects, the promoter is a 117 promoter.

[0029] In some aspects, therefore, the disclosure provides a method for treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17) comprising administering to the subject an effective amount of a nucleic acid comprising or consisting of (a) a nucleotide sequence encoding a U7snRA comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34; or (b) a nucleotide sequence encoding a U7snRNA reverse complement sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 11 , 17, 23, 29, or 35, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 1 , 17, 23, 29, or 35.

[0030] In some aspects, the disclosure provides a method for treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17) comprising administering to the subject an effective amount of a nucleic acid comprising or consisting of (a) a nucleotide sequence comprising at least 80%, atleast 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 42-61 , or (b) a nucleotide sequence comprising any one of SEQ ID NOs: 42-61.

[0031] In some aspects, the disclosure provides a method for treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the D / WD gene (DMD exon 17) comprising administering to the subject an effective amount of a nucleic acid comprising or consisting of (a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 62-161 , or (b) a nucleotide sequence comprising any one of SEQ ID NOs: 62-161 .

[0032] In some aspects, the subject is a human subject. In some aspects, the mutation is any mutation involving, surrounding, or affecting DMD exon 17. In some aspects, the method results in increased expression of dystrophin protein or a functional dystrophin protein in the subject. In some aspects, the method inhibits the progression of dystrophic pathology in the subject. In some aspects, the method improves muscle function in the subject. In some aspects, the improvement in muscle function is an improvement in muscle strength. In some aspects, the improvement in muscle function is an improvement in stability in standing and walking.

[0033] The disclosure also provides use of a nucleic acid of the disclosure, a composition of the disclosure, a vector, nanoparticle, extracellular vesicle, or exosome of the disclosure in treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17). In some aspects, the vector is an AAV. In some aspects, the nucleic acid is a nucleic acid that binds or is complementary to a polynucleotide encoding (a) exon 17 of the DMD gene comprising intronic sequence surrounding exon 17 comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 or 2; (b) exon 17 of the DMD gene or its reverse complementary sequence comprising or consisting of SEQ ID NO: 3 or 4; or (c) the amino acid sequence of SEQ ID NO: 5. In some aspects, the nucleic acid is a nucleic acid comprising: (a) an antisense encoding nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 , or comprising or consisting of the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 ; (b) an antisense encoding nucleotide sequence that binds to a target nucleotide sequence of SEQ ID NO: 6, 12, 18, 24, or 30; (c) an antisense encoding nucleotide sequence that binds to a target mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, atleast 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32; or (d) an antisense encoding nucleotide sequence that binds to a target antisense mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 9, 15, 21 , 27, or 33, or comprising or consisting of the nucleotide sequence of 9, 15, 21 , 27, or 33. In some aspects, the nucleic acid further comprises a promoter and / or enhancer. In some aspects, the nucleic acid comprises two or more promoters and / or enhancers. In some aspects, the promoter is a LI6, U7, tRNA, H1 , CMV, minimal CMV, T7, EF1 -alpha, Minimal EF1 -alpha, or a tissue-specific promoter including, but not limited to, a muscle-specific promoter or a cardiac-specific promoter. In some aspects, the promoter is U6 or H1 . In some aspects, the muscle-specific promoter is unc45b, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, alpha-myosin heavy chain enhancer- / MCK enhancerpromoter (MHCK7), or CK1 . In some aspects, the cardiac-specific promoter is alpha-myosin heavy chain enhancer- / MCK enhancer-promoter (MHCK7), the 250-bp fragment of the myosin light chain-2v (MLC-2v) gene promoter (MLC250), cardiac troponin T (cTnT) promoter, the a- myosin heavy chain (a-MHC) promoter, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, or CK1 . In some aspects, the promoter is a U7 promoter. In some aspects, the enhancer is the alpha-myosin heavy chain (aMHC) enhancer (oMHCEnh) or CMV enhancer. In some aspects, the nucleic acid comprises multiple LI7 promoters and / or multiple alpha-myosin heavy chain (aMHC) enhancers (aMHCEnh) or CMV enhancers.

[0034] In some aspects, therefore, the disclosure provides use of (a) a nucleotide sequence encoding a U7snRA comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34; or (b) a nucleotide sequence encoding a U7snRNA reverse complement sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 11 , 17, 23, 29, or 35, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 1 , 17, 23, 29, or 35 in treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17).

[0035] In some aspects, the disclosure provides use of (a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, atleast 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 42- 61 , or (b) a nucleotide sequence comprising any one of SEQ ID NOs: 42-61 in treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17).

[0036] In some aspects, the disclosure provides use of (a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 62- 161 , or (b) a nucleotide sequence comprising any one of SEQ ID NOs: 62-161 in treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17).

[0037] In some aspects, the use results in increased expression of dystrophin protein or increased expression of an altered form of dystrophin protein which has functional activity of the dystrophin protein. In some aspects, the use inhibits the progression of dystrophic pathology. In some aspects, the use improves muscle function. In some aspects, the improvement in muscle function is an improvement in muscle strength. In some aspects, the improvement in muscle function is an improvement in stability in standing and walking.

[0038] Other features and advantages of the disclosure will become apparent from the following description of the drawing and the detailed description. It should be understood, however, that the drawing, detailed description, and the specific examples, while indicating embodiments of the disclosed subject matter, are given by way of illustration only, because various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description.BRIEF DESCRIPTION OF THE DRAWING

[0039] Fig. 1 A-F shows the design of 1stand 2ndgeneration U7 cassettes of the disclosure.

[0040] Fig. 2A-E shows rAAV1 .U7snRNA-induced DMD exon 17 exclusion in vitro and in vivo. Fig. 2A shows RT-PCR agarose gel images showing exon 17 skipping in two patient- derived FM cells harboring deletions of exons 18-33 or exons 18-41 . Cells were treated with three rAAV1 vectors — AAV.U7.17SAS, AAV.U7.17ESE1 or AAV.U7.17SDS — at four doses from 1 E3 to 1 E6 vg / cell. After 14 days of differentiation, total RNA was extracted, and RT-PCR was performed to detect the level of exon 17 exclusion. Red arrows at the left of the image represent PCR amplicons containing either out-of-frame (OOF) of exons 18-33 and 18-41 , or in-frame (IF) deletions of exons 17-33 and 17-41 . Untreated FM and wild-type (WT) cells carrying a normalDMD transcript were used as controls. The expected size of the WT dystrophin mRNA transcript is 2897 bp, resulting in its insufficient amplification in WT cells under PCR conditions optimized for exon 17 skipping in both FM cells. Fig. 2B shows RT-PCR quantification analysis in FM cells detected the highest levels of exon 17 exclusion at the dose of 1 E6 vg / cell. Fig. 2C shows RT- PCR quantification analysis of exon 17 skipping in DMDDel18 41mice intramuscularly (IM) injected into both left and right sides of tibialis anterior (TA) and gastrocnemius (Gast) muscles with the same rAAV1 vectors at the dose of 5E1 1 vg per muscle. Animals were euthanized 4 weeks post-injection with the rAAV1 vectors. The analysis revealed exclusion of exon 17 when AAV.U7.17SAS and AAV.U7.17SDS were injected in both TA and Gast muscles. The triceps tissue from Dup2Del18-41 (D2A18-41 ) and TA from diluent-treated DMDDel18-41mice were used as controls. D2A18-41 mice were identified from a Dup2 colony of mice which had a spontaneous mutation in the DMD gene and showed a complete absence of dystrophin protein (Gushchina et al., Mol Ther Methods Clin Dev, Volume 31, 101144 (2023); PMID: 38027058 ] Fig. 2D shows representative western blot (WB) images of left and right TA and Gast showing dystrophin expression in DMDDel18-41treated mice 4 weeks post-injection (p.i .). The right six lanes of every blot contain a 6-point standard curve of pooled C57BI / 6 (BI6) mouse (n = 3) lysate diluted in Dup2Del18-41 (D2A18-41) dystrophin-null muscle lysate, ranging from 0% to 50%. Fig. 2E shows quantification analysis of normalized dystrophin levels in treated DMDDel18-41mice. Dystrophin signals were quantified by densitometric analysis using Image Lab software (BioRad, version 6.0.0, build 25). A linear regression curve fit to the calibration curve on each gel was calculated using the 0%— 10% wild-type dystrophin points, and individual samples were quantified against that curve and presented as percentage of dystrophin expression. The acceptance criterion for a given blot was defined as R2> 0.8972. Data are presented here as mean ± SEM (n=4) with individual points. Statistical analysis was performed using one-way ANOVA with the Sidak’s multiple comparisons test: **p < 0.01 .DETAILED DESCRIPTION

[0041] The disclosure provides products, methods, and uses for inducing skipping of exon 17 of the DMD gene in a cell or for treating, ameliorating, delaying the progression of, and / or preventing a muscular dystrophy involving a mutation involving, surrounding, or affecting DMD exon 17. Thus, the disclosure provides products and methods for a therapy designed to induce exon 17 exclusion for the treatment of DMD patients carrying skip-amenable mutations (duplications and deletions) that flank exon 17 of the DMD gene. The dystrophin gene, or the DMD gene, is the largest known human gene, provides instructions for making a protein calleddystrophin. The DMD gene is 2.4 million base-pairs in size, comprises 79 exons and takes over 16 hours to be transcribed and cotranscriptionally spliced.

[0042] Dystrophin is located primarily in muscles used for movement (skeletal muscles) and in heart (cardiac) muscle. Dystrophin not only protects the sarcolemma from eccentric contractions, but also anchors a number of signaling proteins in close proximity to sarcolemma. Muscular dystrophy, including Duchenne’s muscular dystrophy (DMD) is caused by mutations in the dystrophin gene leading to absence of dystrophin protein (427 KDa) in skeletal and cardiac muscles, as well as the gastrointestinal tract and retina. Many clinical cases of DMD are linked to deletion mutations in the DMD gene. In contrast to the deletion mutations, DMD exon duplications account for around 5% of disease-causing mutations in unbiased samples of dystrophinopathy patients [Dent et al., Am J Med Genet, 134(3): 295-298 (2005)], although in some catalogues of mutations the number of duplications is higher, including that published by the United Dystrophinopathy Project by Flanigan et al. [Hum Mutat, 30(12): 1657-1666 (2009)], in which it was 11%.

[0043] Another form of MD is Becker Muscular Dystrophy (BMD). BMD, like DMD, is a genetic disorder that gradually makes the body's muscles weaker and smaller. BMD affects the muscles of the hips, pelvis, thighs, and shoulders, as well as the heart, but is known to cause less severe problems than DMD. BMD is also caused by a change in the dystrophin gene, which makes the protein too short. The flawed dystrophin puts muscle cells at risk for damage with normal use. See also, U.S. Patent Application Publication Nos. 2012 / 0077860, published March 29, 2012; 2013 / 0072541 , published March 21 , 2013; and 2013 / 0045538, published February 21 , 2013.

[0044] Despite many lines of research following the identification of the dystrophin gene, treatment options are limited. The most advanced therapies include those that aim at restoration of the missing protein, dystrophin, using mutation-specific genetic approaches, such as antisense oligonucleotide (AON)-mediated exon skipping. The disclosure provides an approach to treating patients carrying exon 17 skip-amenable mutations including, but not limited to, mutations that are within or that flank exon 17 of the DMD gene. Such mutations are known to affect up to 5.5% of all DMD patients.

[0045] More particularly, the disclosure provides nucleic acids comprising sequences designed to bind DMD exon 17 or DMD exon 17 and its surrounding intronic sequence to provide an altered form of dystrophin protein for use in treating a muscular dystrophy resultingfrom a mutation involving, surrounding, or affecting DMD exon 17. The disclosure provides nucleic acids comprising nucleotide sequences encoding and comprising U7-based small nuclear ribonucleic acids (snRNAs) (U7 snRNAs), and vectors, such as recombinant adeno- associated virus (rAAV), comprising the nucleic acids to deliver nucleic acids encoding 117- based snRNAs to induce exon-skipping of DMD exon 17 to provide an altered form of dystrophin protein for use in treating a muscular dystrophy resulting from a mutation involving, surrounding, or affecting DMD exon 17. Exon skipping is a treatment approach to correct and restore production of dystrophin. For specific genetic mutations it allows the body to make a shorter, usable form of dystrophin, or functional dystrophin. Although up to now exon skipping is not a cure for DMD, it may make the effects of DMD less severe.

[0046] Thus, the disclosure provides nucleic acids for treating any mutation amenable to antisense-mediated exon 17 skipping. In some aspects, such mutation amenable to exon 17 skipping is a mutation involving, surrounding, or affecting DMD exon 17. The rationale of antisense-mediated exon skipping is to induce the skipping of a target exon to restore the reading frame.

[0047] The disclosure provides a nucleic acid (or a nucleic acid molecule) or nucleic acids comprising or consisting of an antisense nucleotide sequence designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the open-reading frame of the DMD gene and result in functional dystrophin protein expression or restoration. Such nucleic acid(s) of the disclosure are provided to target the sequences set out in Table 1 as provided below, e.g., a sequence targeting exon 17, i.e., any of SEQ ID Nos: 1 -4, or a sequence encoding the amino acid sequence of SEQ ID NO: 5. Human exon 17 of the DMD gene with surrounding intronic sequence comprises the nucleotide sequence set out in SEQ ID NO: 1 . Inverted or reverse complement of exon 17 of the DMD gene with surrounding intronic sequence comprises the nucleotide sequence set out in SEQ ID NO: 2. Exon 17 of the DMD gene comprises the nucleotide sequence set out in SEQ ID NO: 3. Inverted or reverse complement of exon 17 of the DMD gene comprises the nucleotide sequence set out in SEQ ID NO: 4. Exon 17 of the DMD gene encodes the amino acid sequence set out in SEQ ID NO: 5.

[0048] In various aspects, the methods of the disclosure also target isoforms and variants of the nucleotide sequence set forth in SEQ ID NO: 1 or 3, or the nucleotide sequence encoding the amino acid sequence set out in SEQ ID NO: 5. In some aspects, the variants comprise 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%,83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, and 70% identity to the nucleotide sequence set forth in SEQ ID NO: 1 or 3 or a nucleotide sequence encoding the amino acid sequence set out in SEQ ID NO: 5. Table 1 provides the sequences of human DMD exon 17 and its surrounding intronic sequence.

[0049] Table 1 . Human DMD Exon 17 - Polynucleotide and Amino Acid Sequences.

[0050] The disclosure includes various nucleic acid comprising target sequences of various regions in and around exon 17, including the sense and antisense sequences set out in Table 1 , and their use in a method for inducing skipping of exon 17 of the D / WD gene in a cell or in amethod for treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17). In some aspects, the subject is a human subject. In some aspects, the muscular dystrophy is DMD. In some aspects, the muscular dystrophy is BMD.

[0051] The disclosure provides a nucleic acid (or a nucleic acid molecule) or nucleic acids (or nucleic acid molecules) comprising or consisting of an antisense encoding U7 snRNA nucleotide sequence (or the sense sequence thereof) designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the open-reading frame of the DMD gene and result in functional dystrophin protein expression or restoration. In some aspects, the nucleic acid comprises any of the nucleotide / polynucleotide sequences of any one of SEQ ID NOs: 6-161. In some aspects, the nucleic acid comprises a variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91 %, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71 %, and 70% identity to any of the polynucleotide / nucleotide sequences of any one of SEQ ID NOs: 6-161 . In some aspects, percent identity is percent sequence identity. In some aspects, identity or percent identity is over the full-length nucleotide sequence. In some aspects, identity or percent identity is over a portion of the sequence. In some aspects, the variant comprises the same biological activity as the non-variant polynucleotide sequence and, thus, functions the same as the non-variant in targeting exon 17 of the DMD gene and restoring the open-reading frame of the DMD gene to express or restore the expression of a functional dystrophin protein. In some aspects, the variant or nucleotide sequence variant comprises sufficient biological activity to target exon 17 of the DMD gene and restore the open-reading frame of the DMD gene and / or result in functional dystrophin protein expression or restoration.

[0052] The disclosure also provides a nucleic acid (or a nucleic acid molecule) or nucleic acids (or nucleic acid molecules) comprising or consisting of an antisense encoding nucleotide sequence designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the openreading frame of the DMD gene and result in functional dystrophin protein expression or restoration. More specifically, such nucleic acid comprises any of the polynucleotide sequences of SEQ ID Nos: 7, 13, 19, 25, or 31 . In some aspects, the nucleic acid comprises a combination of two or more, three or more, four or more, five or more, or six or more of any of the polynucleotide sequences of SEQ ID Nos: 7, 13, 19, 25, or 31 . In some aspects, the nucleicacid comprises a variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, and 70% identity to any of the polynucleotide sequences of SEQ ID NOs: 7, 13, 19, 25, or 31 . See Table 2 provided below.

[0053] The disclosure also provides a nucleic acid (or a nucleic acid molecule) or nucleic acids (or nucleic acid molecules) comprising or consisting of an antisense encoding U7 snRNA nucleotide sequence designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the open-reading frame of the DMD gene and result in functional dystrophin protein expression or restoration. More specifically, such nucleic acid comprises any of the polynucleotide sequences of SEQ ID NOs: 10, 16, 22, 28, or 34. In some aspects, the nucleic acid comprises a combination of two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or 10 or more of any of the polynucleotide sequences of SEQ ID Nos: 10, 16, 22, 28, or 34. In some aspects, the nucleic acid comprises a variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, and 70% identity to any of the polynucleotide sequences of SEQ ID NOs: 10, 16, 22, 28, or 34. See Table 2 provided below.

[0054] The disclosure also provides a nucleic acid (or a nucleic acid molecule) or nucleic acids (or nucleic acid molecules) comprising or consisting of an inverted or reverse complement U7 snRNA nucleotide sequence designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the open-reading frame of the DMD gene and result in functional dystrophin protein expression or restoration. More specifically, such nucleic acid comprises any of the polynucleotide sequences of SEQ ID NOs: 11 , 17, 23, 29, or 35. In some aspects, the nucleic acid comprises a combination of two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or 10 or more of any of the polynucleotide sequences of SEQ ID NOs: 11 , 17, 23, 29, or 35. In some aspects, the nucleic acid comprises a variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91 %, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, and 70% identity to any of the polynucleotide sequences of SEQ ID NOs: 1 1 , 17, 23, 29, or 35. See Table 2 provided below.

[0055] The disclosure also provides a nucleic acid (or a nucleic acid molecule) or nucleic acids (or nucleic acid molecules) comprising or consisting of a nucleotide sequence designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the open-reading frame of the DMD gene and result in functional dystrophin protein expression or restoration. Such target regions comprise or consist of any of the polynucleotide sequences of SEQ ID NOs: 6, 12, 18, 24, or 30. In some aspects, the target region comprises a variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91 %, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71 %, and 70% identity to any of the polynucleotide sequences of SEQ ID NOs: 6, 12, 18, 24, or 30. See Table 2 provided below.

[0056] The disclosure also provides a nucleic acid (or a nucleic acid molecule) or nucleic acids (or nucleic acid molecules) comprising or consisting of a target mRNA nucleotide sequence designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the openreading frame of the DMD gene and result in functional dystrophin protein expression or restoration. More specifically, such nucleic acid comprises any of the polynucleotide sequences of SEQ ID NOs: 8, 14, 20, 26, and 32. In some aspects, the nucleic acid comprises a combination of two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or 10 or more of any of the polynucleotide sequences of SEQ ID NOs: 8, 14, 20, 26, and 32. In some aspects, the nucleic acid comprises a variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91 %, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, and 70% identity to any of the polynucleotide sequences of SEQ ID NOs: 8, 14, 20, 26, and 32. See Table 2 provided below.

[0057] The disclosure also provides a nucleic acid (or a nucleic acid molecule) or nucleic acids (or nucleic acid molecules) comprising or consisting of an antisense RNA nucleotide sequence (or in some instances a sense RNA nucleotide sequence) designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the open-reading frame of the DMD gene and result in functional dystrophin protein expression or restoration. More specifically, such nucleic acid comprises any of the polynucleotide sequences of SEQ ID NOs: 9, 15, 21 , 27, or 33. In some aspects, the nucleic acid comprises a combination of two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or 10 or more ofany of the polynucleotide sequences of SEQ ID NOs: 9, 15, 21 , 27, or 33. In some aspects, the nucleic acid comprises a variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91 %, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71 %, and 70% identity to any of the polynucleotide sequences of SEQ ID NOs: 9, 15, 21 , 27, or 33. See Table 2 provided below showing antisense encoding, antisense, and target sequences of the relevant exon 17 nucleotide sequences of the disclosure. In Table 2, exonic sequence is set out in upper case font and surrounding intronic sequence is set out in lower case font.

[0058] The disclosure also provides a nucleic acid (or a nucleic acid molecule) or nucleic acids (or nucleic acid molecules) comprising or consisting of two or four copies of an antisense or sense encoding U7 snRNA nucleotide sequence designed to target exon 17 of the DMD gene or any of the surrounding regions of exon 17 that affects DMD transcription such that the use of the nucleic acid(s) restore the open-reading frame of the DMD gene and result in functional dystrophin protein expression or restoration. More specifically, such nucleic acid comprises two or four copies of any of the polynucleotide sequences of SEQ ID NOs: 10, 11 , 16, 17, 22, 23, 28, 29, 34, or 35. In some aspects, the nucleic acid comprises a combination of two or more, three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or 10 or more of any of the polynucleotide sequences of SEQ ID SEQ ID NOs: 10, 11 , 16, 17, 22, 23, 28, 29, 34, or 35. In some aspects, the nucleic acid comprises a combination of variant nucleotide sequences comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, and 70% identity to any of the polynucleotide sequences of SEQ ID SEQ ID NOs: 10, 1 1 , 16, 17, 22, 23, 28, 29, 34, or 35. See Table 2 provided below.

[0059] Thus, the disclosure also provides nucleic acids (or nucleic acid constructs or constructs) with two or four copies of U7 antisense or sense sequences. Each of these sequences is driven by the mouse U7 promoter and is designated U7 / AS or U7 / S. In some aspects, each of these U7 / AS or U7 / S sequences is connected by a 12 nucleotide sequence between the copies for cloning purposes. Thus, such construct, in some aspects, comprises the following exemplary structures: 2x copies: (U7 / AS)— 12nt — (U7 / AS) or (U7 / S)-12nt-(U7 / S), or 4x copies: (U7 / AS)— 12nt— (LI7 / AS)— 12nt— (U7 / AS)— 12nt— (U7 / AS) or (U7 / S)— 12nt— (U7 / S)— 12nt — (U7 / S) — 12nt — (U7 / S). More specifically, such nucleic acid comprises any of the polynucleotide sequences of SEQ ID NOs: 41-61 . In some aspects, the nucleic acid comprisesa variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91 %, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81%, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, and 70% identity to any of the polynucleotide sequences of SEQ ID NOs:42-61 . See Table 2 provided below.

[0060] The disclosure also provides a nucleic acid comprising stuffer and / or enhancer sequence in addition to promoter sequence. In some aspects, the inclusion of stuffer and / or enhancer sequence increases efficiency of exon skipping. In some aspects, the nucleic acid further comprises a promoter. In some aspects, the promoter is a U7 promoter. In some aspects, the nucleic acid further comprises multiple promoter. In some aspects, the nucleic acid comprises multiple U7 promoters. In some instances, the nucleic acid further comprises a combination of any two or more of any of the nucleic acids described herein. In some aspects, the combination comprises three or more, four or more, five or more, six or more, seven or more, eight or more, nine or more, or even ten or more nucleic acids of the disclosure. In some aspects, the presence of these additional elements or additional copies of snRNAs in the construct increases efficiency of exon skipping. Such nucleic acid comprises any of the polynucleotide sequences of any one of SEQ ID NOs: 62-161. In some aspects, the nucleic acid comprises a variant comprising 99%, 98%, 97%, 96%, 95%, 94%, 93%, 92%, 91%, 90%, 89%, 88%, 87%, 86%, 85%, 84%, 83%, 82%, 81 %, 80%, 79%, 78%, 77%, 76%, 75%, 74%, 73%, 72%, 71%, and 70% identity to any of the polynucleotide sequences of SEQ ID NOs:62-161 . See Table 2, provided below. Because of the size of the sequences of SEQ ID NOs:62-161 , the sequences are provided herein below in Example 6 and Table 7.

[0061] Table 2. Antisense Encoding, Antisense, and Target Sequences.

[0062] The disclosure provides nucleic acids comprising or consisting of antisense sequences (and sequences that are the reverse complement of the antisense sequences) that interfere with the expression of exon 17 of the DMD gene by interfering with the spliceosome resulting in the skipping of exon 17 of the DMD gene in order to restore the reading frame of the mRNA leading to expression of a truncated dystrophin protein in order to treat, ameliorate and / or prevent a muscular dystrophy resulting from a mutation in the DMD gene and the resultant altered version of mRNA. Thus, as used herein, “increased expression of dystrophin” includes “increased expression of a truncated dystrophin protein, an altered form or dystrophin protein, or a functional fragment of the dystrophin protein” or includes “increased expression of functional dystrophin” or “increased expression of a functional fragment of dystrophin”.

[0063] In some aspects, the disclosure provides nucleic acids comprising any one or more of the sequences set forth in any of SEQ ID NOs: 7, 13, 19, 25, or 31 , or a variant thereof. In some aspects, the disclosure provides nucleic acids comprising any one or more of the sequences set forth in any of SEQ ID NOs: 7, 13, 19, 25, or 31 , or a variant thereof, under the control of a U7 promoter or inserted into a sequence encoding U7 small nuclear RNA (U7 snRNA). Such sequences encoding U7 snRNA are set out in SEQ ID NOs: 10, 16, 22, 28, or 34 (or the reverse complement nucleotide sequences of SEQ ID NOs: 11 , 17, 23, 29, or 35) and can be found in Table 2. In some aspects, multiple copies of these sequences are inserted into the U7-encoding sequences. In some aspects, variants of these sequences are used which may contain one, two, three, four, five, six, seven, eight, nine, or 10 or more nucleotide substitutions, deletions, or insertions. As set out herein above, variants with percent identity to these sequences also are contemplated for use in the disclosure.

[0064] U7 snRNA have been found to be important tools in exon skipping and splicing modulation [Goyenvalle et al., Mol Ther 17(7):1234-40 (2009)]. Moreover, splicing modulation using antisense oligonucleotides (AONs) has been developed for the past two decades as a potential treatment for many diseases, most notably Duchene muscular dystrophy (DMD). Thisincludes pre-clinical and clinical trials [Mendell et al., Ann Neurol 74:637-47 (2013)]. However, such AONs were only shown to mediate weak exon skipping due to the fact that they penetrate the heart and diaphragm (i.e., the most affected muscles in DMD boys) only weakly and they are not stable, i.e., requiring reinjection of DMD patients. It is therefore described herein that AAV-based U7 snRNA gene therapy approaches help circumvent the aforementioned potential delivery problems of AONs.

[0065] The disclosure includes nucleic acids comprising or consisting of the nucleotide sequences encoding U7 snRNA (U7 snRNA antisense sequences, i.e., SEQ ID NOs: 10, 16, 22, 28, or 34, and reverse complement U7 snRNA antisense sequences, i.e., SEQ ID NOs: 1 1 , 17, 23, 29, or 35), that interfere with the expression of exon 17 of the DMD gene by interfering with the spliceosome resulting in the skipping of exon 17 of the DMD gene in order to restore the reading frame of the mRNA leading to expression of a truncated yet functional dystrophin protein in order to treat, ameliorate and / or prevent a muscular dystrophy resulting from a mutation in the DMD gene and the resultant altered version of mRNA. See Table 2.

[0066] In some aspects, the disclosure uses U7 snRNA molecules comprising the nucleotide sequences described herein to inhibit or interfere with splicing. U7 snRNA is normally involved in histone pre-mRNA 3' end processing but, in some aspects, it is converted into a versatile tool for splicing modulation or as antisense RNA that is continuously expressed in cells [Goyenvalle et al., Science 306(5702): 1796-9 (2004)]. By replacing the wild-type LI7 Sm binding site with a consensus sequence derived from spliceosomal snRNAs, the resulting RNA assembles with the seven Sm proteins found in spliceosomal snRNAs. As a result, this U7 Sm OPT RNA accumulates more efficiently in the nucleoplasm and no longer mediates histone pre-mRNA cleavage, although it can still bind to histone pre-mRNA and act as a competitive inhibitor for wild-type U7 small nuclear ribonucleoproteins (snRNPs). By further replacing the sequence binding to the histone downstream element with one complementary to a particular target in a splicing substrate, it is possible to create U7 snRNAs capable of modulating specific splicing events. One advantage of using U7 derivatives is that the antisense sequence is embedded into a small nuclear ribonucleoprotein (snRNP) complex. Moreover, when embedded into a gene therapy vector, these small RNAs can be permanently expressed inside the target cell after a single injection and their use using an AAV approach has been investigated in vivo [Levy et al., EurJ Hum Genet 18(9): 969-70 (2010); Wein et al., Hum Mutat31(2) 136-42 (2010); Wein et al., Nat Med 20(9): 992-1000 (2014)].

[0067] There are three major features to the U7-snRNA system: the U7 promoter to drive expression of (1 ) the modified snRNA in target cells; (2) an antisense sequence inserted in the snRNA backbone, which is designed to base-pair with splice junctions, branch points, or splicing enhancers; (3) a modified sequence (called smOPT) which recruits a distinct ring of RNA binding proteins that complexes with the U7snRNA making it more stable. [Schumperli et al., Cell and Mol Life Sciences 61 :2560-70 (2004)]. It is noteworthy that the antisense sequence and the U7 small nuclear RNA (snRNA) (117 snRNA) have proven safe for use in vivo in large animal models of muscular dystrophy [LeGuiner et al., Mol Ther 22.1923-35 (2014)].

[0068] The disclosure includes nucleic acids that bind to or are complementary to a polynucleotide encoding exon 17 of the DMD gene comprising intronic sequence surrounding exon 17 comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 or 2. The disclosure includes nucleic acids that bind to or are complementary to a polynucleotide encoding exon 17 of the DMD gene or its reverse complementary sequence comprising or consisting of SEQ ID NO: 3 or 4. The disclosure includes nucleic acids that bind to or are complementary to a polynucleotide encoding the amino acid sequence of SEQ ID NO: 5.

[0069] The disclosure includes nucleic acids comprising or consisting of an antisense encoding nucleotide sequence comprising or having at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NOs: 7, 13, 19, 25 or 31.

[0070] The disclosure includes nucleic acids comprising or consisting of an antisense encoding nucleotide sequence that binds to a target nucleotide sequence of SEQ ID NO: 6, 12, 18, 24, or 30.

[0071] The disclosure includes nucleic acids comprising or consisting of an antisense encoding nucleotide sequence that binds to a target mRNA sequence comprising or having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NOs: 8, 14, 20, 26, or 32.

[0072] The disclosure includes nucleic acids comprising or consisting of an antisense encoding nucleotide sequence that binds to a target antisense mRNA sequence comprising orhaving at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NOs: 9, 15, 21 , 27, or 33.

[0073] The disclosure includes nucleic acids comprising or consisting of a nucleotide sequence encoding a U7snRA comprising or having at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NO: 10, 16, 22, 28, or 34.

[0074] The disclosure includes nucleic acids comprising or consisting of a nucleotide sequence encoding a U7snRA reverse complement sequence comprising or having at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NO: 1 1 , 17, 23, 29, or 35.

[0075] The disclosure includes a nucleic acid comprising a combination of at least two of the nucleic acids of the disclosure. In some aspects, the nucleic acid comprises a combination of at least 3 copies of a nucleotide sequence, at least 4 copies of a nucleotide sequence, at least 5 copies of a nucleotide sequence, at least 6 copies of a nucleotide sequence, at least 7 copies of a nucleotide sequence, at least 8 copies of a nucleotide sequence, at least 9 copies of a nucleotide sequence, or at least 10 copies of a nucleotide sequence present in a nucleic acid of the disclosure. For example, in some aspects, the efficiency of exon skipping is shown to be enhanced by delivering multiple copies, i.e., at least two or more copies, of a nucleic acid comprising a nucleic acid of the disclosure. In some aspects, the efficiency of exon skipping is shown to be enhanced by delivering four or more copies of a nucleic acid comprising a nucleic acid of the disclosure.

[0076] In some aspects, exon skipping efficiency is improved by delivering at least two copies of an antisense encoding nucleotide sequence comprising or having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to thenucleotide sequence set out in any of SEQ ID NOs: 7, 13, 19, 25 or 31. In some aspects, exon skipping efficiency is improved by delivering at least four copies of such nucleotide sequence.

[0077] In some aspects, exon skipping efficiency is improved by delivering at least two copies of an antisense encoding nucleotide sequence that binds to a target nucleotide sequence of SEQ ID NO: 6, 12, 18, 24, or 30. In some aspects, exon skipping efficiency is improved by delivering at least four copies of such nucleotide sequence.

[0078] In some aspects, exon skipping efficiency is improved by delivering at least two copies of an antisense encoding nucleotide sequence that binds to a target mRNA sequence comprising or having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NOs: 8, 14, 20, 26, or 32. In some aspects, exon skipping efficiency is improved by delivering at least four copies of such nucleotide sequence.

[0079] In some aspects, exon skipping efficiency is improved by delivering at least two copies of an antisense encoding nucleotide sequence that binds to a target antisense mRNA sequence comprising or having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NOs: 9, 15, 21 , 27, or 33. In some aspects, exon skipping efficiency is improved by delivering at least four copies of such nucleotide sequence.

[0080] In some aspects, exon skipping efficiency is improved by delivering at least two copies of a nucleotide sequence encoding a U7snRA comprising or having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NO: 10, 16, 22, 28, or 34. In some aspects, exon skipping efficiency is improved by delivering at least four copies of such nucleotide sequence.

[0081] In some aspects, exon skipping efficiency is improved by delivering at least two copies of a nucleotide sequence encoding a U7snRA reverse complement sequence comprising or having at least 80%, at least 81 %, at least 82%, at least 83%, at least 84%, at least 85%, atleast 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the nucleotide sequence set out in any of SEQ ID NO: 11 , 17, 23, 29, or 35.

[0082] Thus, the disclosure provides nucleic acids, including nucleic acids encoding target sequence, nucleic acids encoding antisense sequences and reverse complements of the antisense sequences, nucleic acids encoding U7-based small nuclear ribonucleic acids (snRNAs), i.e., U7-based snRNAs, nucleic acids encoding the reverse complement of the 117- based snRNAs, and vectors (including but not limited to AAV or rAAV), nanoparticles, extracellular vesicles, or exosomes comprising the nucleic acids to induce exon-skipping for use in treating a muscular dystrophy. In some aspects, the disclosure includes nucleic acids, sometimes referred to herein as exon 17 U7 snRNA polynucleotide constructs, or exon 17- targeted U7 snRNA, which inhibit or interfere with the expression and / or incorporation of exon 17 of the DMD gene into the mRNA. The disclosure contemplates that the nucleic acids encoding these inhibitory splicing RNAs are responsible for sequence-specific gene exon skipping.

[0083] In some embodiments, a nucleic acid of the disclosure further comprise a promoter. In some aspects, the promoter sequence is already part of the nucleic acid. In some embodiments of the disclosure, the promoter DNAs are muscle-specific control elements, including, but not limited to, those derived from the actin and myosin gene families, such as from the myoD gene family [See Weintraub et al., Science, 251 : 761 -766 (1991 )], the myocytespecific enhancer binding factor MEF-2 [Cserjesi and Olson, Mol. Cell. Biol., 11 : 4854-4862 (1991 )], control elements derived from the human skeletal actin gene [Muscat et al., Mol. Cell. Biol., 7: 4089-4099 (1987)], the cardiac actin gene, muscle creatine kinase sequence elements [Johnson et al., Mol. Cell. Biol., 9:3393-3399 (1989)] and the murine creatine kinase enhancer (MCK) element, desmin promoter, control elements derived from the skeletal fast-twitch troponin C gene, the slow-twitch cardiac troponin C gene and the slow-twitch troponin I gene: hypozia- inducible nuclear factors [Semenza et al., Proc. Natl. Acad. Sci. USA, 88: 5680-5684 (1991 )], steroid-inducible elements and promoters including the glucocorticoid response element (GRE) [See Mader and White, Proc. Natl. Acad. Sci. USA, 90: 5603-5607 (1993)], and other control elements.

[0084] In some aspects, the promoter is any of U6, U7, tRNA, H1 , CMV, minimal CMV, T7, EF1 -alpha, Minimal EF1 -alpha, or a tissue-specific promoter including, but not limited to, a muscle-specific promoter or a cardiac-specific promoter. In some aspects, the promoter is U6or H1 . In some aspects, the muscle-specific promoter is unc45b, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, alpha-myosin heavy chain enhancer- / MCK enhancerpromoter (MHCK7), or CK1 . In some aspects, the cardiac-specific promoter is alpha-myosin heavy chain enhancer- / MCK enhancer-promoter (MHCK7), the 250-bp fragment of the myosin light chain-2v (MLC-2v) gene promoter (MLC250), cardiac troponin T (cTnT) promoter, the ?- myosin heavy chain (a-MHC) promoter, muscle creatine kinase (MCK), tMCK, minimal MCK, CK6, CK7, CK8, or CK1 . In some exemplary aspects, the promoter is a U7 promoter, but the disclosure is not limited to the use of only a U7 promoter, and other promoters are disclosed herein.

[0085] In some aspects, the nucleic acid (or nucleic acids) is delivered in a vector. In some aspects, the vector is an AAV vector. In some aspects, the nucleic acid (or nucleic acids) is delivered in a nanoparticle, extracellular vesicle, or exosome. In some aspects, a combination of nucleic acids is delivered in a vector, nanoparticle, extracellular vesicle, or exosome. In some aspects, any of the nucleic acids, vectors, AAV, or nanoparticles, extracellular vesicles, or exosomes are present in a composition or a pharmaceutical composition described herein.

[0086] In some aspects, the vectors are viral vectors, such as adeno-associated virus (AAV), adenovirus, retrovirus, lentivirus, equine-associated virus, alphavirus, pox virus, herpes virus, herpes simplex virus, polio virus, sindbis virus, vaccinia virus or a synthetic virus, e.g., a chimeric virus, mosaic virus, or pseudotyped virus, and / or a virus that contains a foreign protein, synthetic polymer, nanoparticle, or small molecule) to deliver the nucleic acids disclosed herein. In some aspects, adeno-associated virus (AAV) is used. In some aspects, recombinant adeno- associated virus (rAAV) is used.

[0087] AAV is a replication-deficient parvovirus, the single-stranded DNA genome of which is about 4.7 kb in length including two 145 nucleotide inverted terminal repeat (ITRs) and the double-stranded DNA genome of which is about 2.3 kb in length, including two 145 nucleotide ITRs. There are multiple serotypes of AAV. The nucleotide sequences of the genomes of the AAV serotypes are known. For example, the complete genome of AAV1 is provided in Gen Bank Accession No. NC_002077; the complete genome of AAV2 is provided in Gen Bank Accession No. NC 001401 and Srivastava et al., J Virol, 45: 555-64 (1983); the complete genome of AAV3 is.provided in GenBank Accession No. NC_1829; the complete genome of AAV4 is provided in GenBank Accession No. NC_001829; the AAV5 genome is provided in GenBank Accession No. AF085716; the complete genome of AAV6 is provided in GenBank Accession No. NC_00 1862; at least portions of AAV7 and AAV8 genomes are provided inGenBank Accession Nos. AX753246 and AX753249, respectively; the AAVrh74 genome; the AAV9 genome is provided in Gao et al., J Virol, 78: 6381-8 (2004); the AAV10 genome is provided in Mol Ther 13(1): 67-76 (2006); the AAV-11 genome is provided in Virology, 330(2): 375-83 (2004); the genome of AAV12 is provided in GenBank Accession No. DQ813647.1 ; and the genome of AAV13 is provided in GenBank Accession No. EU285562.1. A full list of all known AAV genomes is not provided herein; however, additional AAV vectors / genomes are known in the art and are considered to be generally contemplated for use in delivering any of the nucleic acids of the disclosure.

[0088] C / s-acting sequences directing viral DNA replication (rep), encapsidation / packaging and host cell chromosome integration are contained within the AAV ITRs. Three AAV promoters (named p5, p19, and p40 for their relative map locations) drive the expression of the two AAV internal open reading frames encoding rep and cap genes. The two rep promoters (p5 and p19), coupled with the differential splicing of the single AAV intron (at nucleotides 2107 and 2227), result in the production of four rep proteins (rep 78, rep 68, rep 52, and rep 40) from the rep gene. Rep proteins possess multiple enzymatic properties that are ultimately responsible for replicating the viral genome. The cap gene is expressed from the p40 promoter and it encodes the three capsid proteins VP1 , VP2, and VP3. Alternative splicing and non-consensus translational start sites are responsible for the production of the three related capsid proteins. A single consensus polyadenylation site is located at map position 95 of the AAV genome. The life cycle and genetics of AAV are reviewed in Muzyczka, Current Topics in Microbiology and Immunology, 158: 97-129 (1992).

[0089] AAV possesses unique features that make it attractive as a vector for delivering foreign DNA to cells, for example, in gene therapy. AAV infection of cells in culture is noncytopathic, and natural infection of humans and other animals is silent and asymptomatic. Moreover, AAV infects many mammalian cells allowing the possibility of targeting many different tissues in vivo. Moreover, AAV transduces slowly dividing and non-dividing cells, and can persist essentially for the lifetime of those cells as a transcriptionally active nuclear episome (extrachromosomal element). The AAV proviral genome is inserted as cloned DNA in plasmids which makes construction of recombinant genomes feasible. Furthermore, because the signals directing AAV replication and genome encapsidation are contained within the ITRs of the AAV genome, some or all of the internal approximately 4.3 kb of the genome (encoding replication and structural capsid proteins, rep-cap) may be replaced with foreign DNA. To generate AAV vectors, the rep and cap proteins may be provided in trans. Another significant feature of AAVis that it is an extremely stable and hearty virus. It easily withstands the conditions used to inactivate adenovirus (56° to 65°C for several hours), making cold preservation of AAV less critical. AAV may even be lyophilized. Finally, AAV-infected cells are not resistant to superinfection.

[0090] Recombinant AAV, i.e. , rAAV genomes, of the disclosure comprise one or more AAV ITRs flanking at least one exon 17-targeted U7 snRNA polynucleotide construct. Genomes with exon 17-targeted U7 snRNA polynucleotide constructs comprising each of the exon 17 targeting antisense sequences as described herein are specifically contemplated, as well as genomes with exon 17-targeted U7 snRNA polynucleotide constructs comprising each possible combination of two or more of the exon 17 targeting antisense sequences described herein. In some embodiments, the U7 snRNA polynucleotide includes its own promoter. In some aspects, rAAV of the disclosure comprise one or more AAV ITRs flanking a polynucleotide encoding, for example, one or more antisense sequences targeting DMD exon 17 or one or more DMD exon 17 117-based snRNAs (i.e., an snRNA that binds to a gene sequence within or surrounding exon 17 and, in some aspects, is expressed from a U7 snRNA). In some aspects, the polynucleotide is operatively linked to transcriptional control DNA, specifically promoter DNA that is functional in target cells as disclosed herein above.

[0091] AAV DNA in the rAAV genomes may be from any AAV serotype for which a recombinant virus can be derived including, but not limited to, AAV serotypes AAV is AAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV.rh74, AAV.rh8, AAV.rh , AAV1 1 , AAV12, AAV13, AAV-anc80, AAV-B1 , AAV-BR1 , AAV.PHP.EB, AAVv66, AAV2 / 1 , AAV2 / 8, AAV2 / 9, AAVMYO, MYOAAV, MYOAAV1 A, MYOAAV2A, MYOAAV3A, or any other myotropic serotype, or any derivative thereof. The nucleotide sequences of the genomes of these various AAV serotypes are known in the art.

[0092] DNA plasmids of the disclosure comprise rAAV genomes of the disclosure. The DNA plasmids are transferred to cells permissible for infection with a helper virus of AAV (e.g., adenovirus, E1 -deleted adenovirus or herpesvirus) for assembly of the rAAV genome into infectious viral particles. Techniques to produce rAAV particles, in which an AAV genome to be packaged, rep and cap genes, and helper virus functions are provided to a cell are standard in the art. Production of rAAV requires that the following components are present within a single cell (denoted herein as a packaging cell): a rAAV genome, AAV rep and cap genes separate from (i.e., not in) the rAAV genome, and helper virus functions. The AAV rep genes may be from any AAV serotype for which recombinant virus can be derived and may be from a differentAAV serotype than the rAAV genome ITRs, including, but not limited to, AAV serotypes AAV is AAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV.rh74, AAV.rh8, AAV.rh , AAV11 , AAV12, AAV13, AAV-anc80, AAV-B1 , AAV-BR1 , AAV.PHP.EB, AAVv66, AAV2 / 1 , AAV2 / 8, AAV2 / 9, AAVMYO, MYOAAV, MYOAAV1 A, MYOAAV2A, MY0AAV3A, or any other myotropic serotype, or any derivative thereof. Use of cognate components is specifically contemplated. Production of pseudotyped rAAV is disclosed in, for example, WO 01 / 83692 which is incorporated by reference herein in its entirety.

[0093] In some embodiments of the disclosure, the virus genome is a single-stranded genome or a self-complementary genome. In some embodiments of the methods, the genome of the rAAV lacks AAV rep and cap DNA.

[0094] The disclosure also provides a method of generating a packaging cell. The purpose of generating a packing cell is to create a cell line that stably expresses all the necessary components for AAV particle production. For example, a plasmid (or multiple plasmids) comprising a rAAV genome lacking AAV rep and cap genes, AAV rep and cap genes separate from the rAAV genome, and a selectable marker, such as a neomycin resistance gene, are integrated into the genome of a cell. AAV genomes have been introduced into bacterial plasmids by procedures such as GC tailing [Samulski et al., Proc Natl Acad Sci USA, 79:2077- 81 (1982)], addition of synthetic linkers containing restriction endonuclease cleavage sites [Laughlin et al., Gene, 23:65-73 (1983)] or by direct, blunt-end ligation [Senapathy et al., J Biol Chem 259:4661 -6 (1984)]. The packaging cell line is then infected with a helper virus such as adenovirus. The advantages of this method are that the cells are selectable and are suitable for large-scale production of rAAV. Other examples of suitable methods employ adenovirus or baculovirus rather than plasmids to introduce rAAV genomes and / or rep and cap genes into packaging cells.

[0095] General principles of rAAV production are reviewed in, for example, Carter, Current Opinions in Biotechnology, 1533-539 (1992); and Muzyczka, Curr Topics in Microbial and Immunol, 158:97-129 (1992)). Various approaches are described in Ratschin et al., Mol. Cell. Biol. 4:2072 (1984); Hermonat et al., Proc. Natl. Acad. Sci. USA, 81 :6466 (1984); Tratschin et al., Mol. Cell. Biol. 5:3251 (1985); McLaughlin et al., J. Virol., 62:1963 (1988); and Lebkowski et al., Mol. Cell. Biol., 7:349 (1988); Samulski et al., J. Virol., 63:3822-8 (1989); U.S. Patent No. 5,173,414; WO 95 / 13365 and corresponding U.S. Patent No. 5,658.776; WO 95 / 13392; WO 96 / 17947; PCT / US98 / 18600; WO 97 / 09171 (PCT / US96 / 1 1723); WO 97 / 08298(PCT / US96 / 13872); WO 97 / 21825 (PCT / US96 / 20777); WO 97 / 06243 (PCT / FR96 / 01064); WO 99 / 11764; Perrin et al., Vaccine 13:1217-50 (1995); Paul etal., Human Gene Therapy 4:609- 615 (1993); Clark et al., Gene Therapy 3:1124-32 (1996); U.S. Patent. No. 5,786,21 1 ; U.S. Patent No. 5,871 ,982; and U.S. Patent. No. 6,258,595. The foregoing documents are hereby incorporated by reference in their entirety herein, with particular emphasis on those sections of the documents relating to rAAV production.

[0096] The disclosure thus provides packaging cells that produce infectious rAAV. In one embodiment packaging cells may be stably transformed cancer cells such as HeLa cells, 293 cells and PerC.6 cells (a cognate 293 line). In another embodiment, packaging cells are cells that are not transformed cancer cells, such as low passage 293 cells (human fetal kidney cells transformed with E1 of adenovirus), MRC-5 cells (human fetal fibroblasts), WI-38 cells (human fetal fibroblasts), Vero cells (monkey kidney cells) and FRhL-2 cells (rhesus fetal lung cells). Cell transduction efficiencies of the methods of the disclosure described above and below may be at least about 60, 65, 70, 75, 80, 85, 90 or 95 percent efficient.

[0097] The rAAV of the disclosure, in various aspects, are purified by methods standard in the art such as by column chromatography or cesium chloride gradients. Methods for purifying rAAV vectors from helper virus are known in the art and include methods disclosed in, for example, Clark et al., Hum. Gene Ther. 10(6): 1031 -9 (1999); Schenpp et al., Methods Mol. Med. 69:427-43 (2002); U.S. Patent No. 6,566,118; and WO 98 / 09657.

[0098] In another embodiment, the disclosure provides compositions comprising the nucleic acids of the disclosure. In some embodiments, the disclosure provides vectors (including, but not limited to, AAV or rAAV), nanoparticles, extracellular vesicles, or exosomes comprising any of the nucleic acids or nucleic acid constructs described herein. In one aspect, the disclosure includes a composition comprising rAAV for delivering the antisense RNAs or the U7snRNAs described herein. Some compositions of the disclosure comprise rAAV in a pharmaceutically acceptable carrier. The compositions may also comprise other ingredients such as diluents. Acceptable carriers and diluents are nontoxic to recipients and are preferably inert at the dosages and concentrations employed, and include buffers such as phosphate, citrate, or other organic acids; antioxidants such as ascorbic acid; low molecular weight polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and / or nonionic surfactants such as Tween, pluronics or polyethylene glycol (PEG).

[0099] Sterile injectable solutions are prepared by incorporating nucleic acids, vectors (including, but not limited to rAAV vectors), nanoparticles, extracellular vesicles, or exosomes in the required amount in the appropriate solvent with various other ingredients enumerated above, as required, followed by filter sterilization. Generally, dispersions are prepared by incorporating the sterilized active ingredient into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying technique that yield a powder of the active ingredient plus any additional desired ingredient from the previously sterile-f iltered solution thereof.

[0100] In some aspects, when rAAV are used, the rAAV is titered. Titers of rAAV to be administered in methods of the disclosure will vary depending, for example, on the particular rAAV, the mode of administration, the treatment goal, the individual, and the cell type(s) being targeted, and may be determined by methods standard in the art. Titers of rAAV may range from about 1 x106, about 1 x107, about 1 x108, about 1 x109, about 1 x1010, about 1 x1011, about 1 x1012, about 1x1013to about 1 x1014or more DNase resistant particles (DRP) per ml.

[0101] Dosages, in various aspects, are expressed in units of viral genomes (vg) ( / .e., 1 x107vg, 1 x108vg, 1 x109vg, 1x101° vg, 1x1011vg, 1 x1012vg, 1x1013vg, 1 x1014vg, respectively). In some aspects of the disclosure, dosages of rAAV range from about 1 x105, about 1 x106, about 1 x107, about 1 x108, about 1 x109, about 1 x1010, about 1 x1011, about 1 x1012, about 1 x1013to about 1 x1014or more vg / cell. In some aspects of the disclosure, dosages of rAAV range from about 1 x105, about 1 x106, about 1 x107, about 1 x108, about 1x109, about 1 x1010, about 1x1011, about 1x1012, about 1 x1013, about 1 x1014, about 1 x1015, or about 1 x1016or more vg / injection site. In some aspects of the disclosure, dosages of rAAV range from about 1 x105, about 1 x106, about 1 x107, about 1 x108, about 1 x109, about 1 x1010, about 1 x1011, about 1 x1012, about 1 x1013, about 1 x1014, about 1 x1015, or about 1 x1016or more vg / kg.

[0102] The disclosure thus provides a method of delivering a nucleic acid of the disclosure to a subject in need thereof. In some aspects, the method comprises administering to the subject a vector, nanoparticle, extracellular vesicle, or exosome comprising the exon 17- targeted nucleic acids of the disclosure. In some aspects, the vector being administered to the subject is an AAV comprising such exon 17-targeted nucleic acids.

[0103] Methods of transducing a target cell (e.g., a skeletal muscle) with a vector, nanoparticle, extracellular vesicle, or exosome, in vivo or in vitro, are contemplated by the disclosure. The methods comprise the step of administering an effective dose, or effective multiple doses, of a composition comprising a vector, nanoparticle, extracellular vesicle, or exosome of the disclosure to an animal (including a human being) in need thereof. If the dose is administered prior to development of a muscular dystrophy, e.g., DMD or BMD, the administration is prophylactic. If the dose is administered after the development of a muscular dystrophy, the administration is therapeutic. In embodiments of the disclosure, an effective dose is a dose that alleviates (eliminates or reduces) at least one symptom associated with a muscular dystrophy being treated, that slows or prevents progression of the muscular dystrophy, e.g. DMD or BMD, that slows or prevents progression of the muscular dystrophy disorder / disease state, that diminishes the extent of disease, that results in remission (partial or total) of disease, and / or that promotes improved muscle strength and / or prolongs survival of the subject suffering from the disorder or disease.

[0104] Administration of an effective dose of a composition of the disclosure or a nucleic acid, vector, nanoparticle, extracellular vesicle, or exosome of the disclosure may be by routes standard in the art including, but not limited to, intramuscular, parenteral, intravenous, intrathecal, oral, buccal, nasal, pulmonary, intracranial, intraosseous, intraocular, rectal, or vaginal. Route(s) of administration and serotype(s) of AAV components of rAAV (in particular, the AAV ITRs and capsid protein) of the disclosure may be chosen and / or matched by those skilled in the art taking into account the infection and / or disease state being treated and the target cells / tissue(s). In some embodiments, the route of administration is intramuscular, intravenous, or subcutaneous. In some particular embodiments, the route of administration is intramuscular. In particular aspects, the route of administration takes into consideration the target cells / tissue(s) that express the DMD gene and the dystrophin protein.

[0105] In particular, actual administration of a nucleic acid, vector, nanoparticle, extracellular vesicle, or exosome of the disclosure is, in some aspects, accomplished by using any physical method that will transport the nucleic acid, vector, nanoparticle, extracellular vesicle, or exosome into the target tissue of a subject. Administration according to the disclosure includes, but is not limited to, injection into muscle, the liver, the cerebral spinal fluid, or the bloodstream. In some aspects, when administering an rAAV, simply resuspending an rAAV in phosphate buffered saline has been demonstrated to be sufficient to provide a vehicle useful for muscle tissue expression, and there are no known restrictions on the carriers or othercomponents that can be co-administered with the rAAV (although compositions that degrade DNA should be avoided in the normal manner with rAAV). In some aspects, capsid proteins of an rAAV are modified so that the rAAV is targeted to a particular target tissue of interest, such as muscle. See, for example, WO 02 / 053703, the disclosure of which is incorporated by reference herein. In some aspects, compositions or pharmaceutical compositions are prepared as injectable formulations or as topical formulations to be delivered to the muscles by transdermal transport. Numerous formulations for both intramuscular injection and transdermal transport have been previously developed and can be used in the practice of the disclosure. In some aspects, the rAAV are used with any pharmaceutically acceptable carrier or excipient for ease of administration and handling.

[0106] In some aspects, for purposes of intramuscular injection, solutions in an adjuvant, such as sesame or peanut oil or in aqueous propylene glycol, are employed, as well as sterile aqueous solutions. Such aqueous solutions, in various aspects, are buffered, if desired, and the liquid diluent is rendered isotonic with saline or glucose. In some aspects, solutions of rAAV as a free acid (DNA contains acidic phosphate groups) or a pharmacologically acceptable salt are prepared in water, suitably mixed with a surfactant such as hydroxpropylcellulose. In various aspects, a dispersion of rAAV is prepared in glycerol, liquid polyethylene glycol(s) and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. In this connection, the sterile aqueous media employed are all readily obtainable by standard techniques in the art.

[0107] Formulations, including pharmaceutical forms suitable for injectable use, include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating actions of microorganisms, such as bacteria and fungi. In some aspects, the carrier is a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. Proper fluidity, in some aspects, is maintained by the use of a coating, such as lecithin, by the maintenance of the required particle size, in the case of a dispersion, and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In some aspects, it is preferable to include isotonic agents, forexample, sugars or sodium chloride. Prolonged absorption of the injectable compositions, in some aspects, is brought about by use of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0108] Sterile injectable solutions are prepared, in some aspects, by incorporating rAAV in the required amount in the appropriate solvent with various other ingredients enumerated above, as required, followed by filter sterilization. Generally, dispersions are prepared by incorporating the sterilized active ingredient into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, various methods of preparation are vacuum drying and the freeze drying technique that yield a powder of the active ingredient plus any additional desired ingredient from the previously sterile-f iltered solution thereof.

[0109] Transduction with rAAV, in some aspects, is also carried out in vitro. In one embodiment, for example, desired target muscle cells are removed from the subject, transduced with rAAV and reintroduced into the subject. Alternatively, syngeneic or xenogeneic muscle cells, in some aspects, are used where those cells will not generate an inappropriate immune response in the subject.

[0110] Suitable methods for the transduction and reintroduction of transduced cells into a subject are known in the art. In one embodiment, cells are transduced in vitro by combining rAAV with muscle cells, e.g., in appropriate media, and screening for those cells harboring the DNA of interest using conventional techniques in the art, such as Southern blots and / or PGR, or by using selectable markers. Transduced cells, in some aspects, are then formulated into a composition, including a pharmaceutical composition, and the composition is introduced into the subject by various techniques, such as by intramuscular, intravenous, subcutaneous, and / or intraperitoneal injection, or by injection into smooth and cardiac muscle, using e.g., a catheter.

[0111] The disclosure provides methods of administering an effective dose (or doses, administered essentially simultaneously or doses given at intervals) of rAAV that encode inhibitory RNAs and rAAV that encode combinations of inhibitory RNAs, including snRNAs, that target exon 17, and skipping of exon 17, to a subject in need thereof.

[0112] Transduction of cells with rAAV of the invention results in sustained expression of the exon 17 U7-based snRNAs. The term “transduction” is used to refer to the administration / delivery of one or more exon 17-targeted U7snRNA polynucleotide construct to arecipient cell either in vivo or in vitro, via a replication-deficient rAAV of the invention resulting in expression of the one or more exon 17-targeted U7snRNA polynucleotide construct by the recipient cell. The disclosure thus provides methods of administering / delivering rAAV which express exon 17 U7-based snRNAs to a subject. In some aspects, the subject is a human being.

[0113] These methods include transducing the blood and vascular system, the central nervous system, and tissues (including, but not limited to, tissues, such as muscle, organs such as liver and brain, and glands such as salivary glands) with one or more rAAV of the disclosure. Transduction, in some aspects, is carried out with gene cassettes comprising tissue specific control elements. For example, one embodiment of the disclosure provides methods of transducing muscle cells and muscle tissues directed by muscle specific control elements, including, but not limited to, those derived from the actin and myosin gene families, such as from the myoD gene family [See Weintraub et al., Science, 251: 761 -6 (1991 )], the myocytespecific enhancer binding factor MEF-2 [Cserjesi et al., Mol Cell Biol 1 1 : 4854-62 (1991 )], control elements derived from the human skeletal actin gene [Muscat et al., Mol Cell Biol, 7: 4089-99 (1987)], the cardiac actin gene, muscle creatine kinase sequence elements [See Johnson et al., Mol Cell Biol, 9:3393-9 (1989)] and the murine creatine kinase enhancer (mCK) element, control elements derived from the skeletal fast-twitch troponin C gene, the slow-twitch cardiac troponin C gene and the slow-twitch troponin I gene: hypoxia-inducible nuclear factors [Semenza etal., Proc Natl Acad Sci USA, 88: 5680-4 (1991 )], steroid-inducible elements and promoters including the glucocorticoid response element (GRE) [See Mader et al., Proc Natl Acad Sci USA 90: 5603-7 (1993)], and other control elements.

[0114] Because AAV targets every dystrophin affected organ, the disclosure includes the delivery of DNAs encoding the inhibitory RNAs to all cells, tissues, and organs of a subject. In some aspects, the blood and vascular system, the central nervous system, muscle tissue, the heart, and the brain are attractive targets for in vivo DNA delivery. The disclosure includes the sustained expression of snRNA from transduced cells to affect DMD exon 17 expression (e.g., skip, knockdown or inhibit expression) and alter expression of the DMD protein. In some aspects, muscle tissue is targeted for delivery of the nucleic acid molecules and vectors of the disclosure. Muscle tissue is an attractive target for in vivo DNA delivery, because it is not a vital organ and is easy to access. The disclosure, in some aspects, contemplates sustained expression of one or more exon 17 U7-based snRNAs from transduced myofibers. By "muscle cell" or "muscle tissue" is meant a cell or group of cells derived from muscle of any kind (forexample, skeletal muscle and smooth muscle, e.g. from the digestive tract, urinary bladder, blood vessels or cardiac tissue). Such muscle cells, in some aspects, are differentiated or undifferentiated, such as myoblasts, myocytes, myotubes, cardiomyocytes and cardiomyoblasts.

[0115] In yet another aspect, the disclosure provides a method of restoring the open reading frame of the DMD gene in a cell comprising contacting the cell with a nucleic acid or a vector, nanoparticle, extracellular vesicle, or exosome comprising the nucleic acid encoding an exon 17-targeted antisense of the disclosure or the nucleic acid encoding an exon 17-targeted U7 snRNA of the disclosure. In some aspects, skipping of exon 17 results in exclusion or inhibition of exon 17 by at least about 5, about 10, about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 96, about 97, about 98, about 99, or 100 percent.

[0116] Thus, the disclosure provides methods of administering an effective dose (or doses, administered essentially simultaneously or doses given at intervals) of an exon 17-targeted U7snRNA polynucleotide construct or an rAAV that comprises a genome that encodes one or more exon 17-targeted U7snRNA polynucleotide construct to a subject in need thereof e.g., a subject or patient suffering from a muscular dystrophy, such as DMD).

[0117] In some aspects, a method of treating muscular dystrophy in a patient is provided. In some aspects, “treating” includes ameliorating, inhibiting, or even preventing one or more symptoms of a muscular dystrophy, including a Duchenne muscular dystrophy, (including, but not limited to, muscle wasting, muscle weakness, skeletal muscle problems, heart function abnormalities, breathing difficulties, issues with speech and swallowing (dysarthria and dysphagia) or cognitive impairment). In some aspects, the method of treating results in increased expression of dystrophin protein or increased expression of an altered form or fragment of dystrophin protein that is physiologically or functionally active in the subject. In particular aspects, the method of treating inhibits the progression of dystrophic pathology in the subject. In some aspects, the method of treating improves muscle function in the subject. In some aspects, the improvement in muscle function is an improvement in muscle strength. In some aspects, the improvement in muscle function is an improvement in stability in standing and walking. The improvement in muscle strength is determined by techniques known in the art, such as the maximal voluntary isometric contraction testing (MVICT). In some instances, the improvement in muscle function is an improvement in stability in standing and walking. Insome aspects, an improvement in stability or strength is determined by techniques known in the art such as the 6-minute walk test (6MWT), the 100 meter run / walk test, or timed stair climb.

[0118] In some embodiments, the method of treating comprises the step of administering one or more exon 17 U7-based snRNA polynucleotide construct without the use of a vector. In some embodiments, the method of treating comprises the step of administering an rAAV to the subject, wherein the genome of the rAAV comprises one or more exon 17 U7-based snRNA polynucleotide construct.

[0119] In yet another aspect, the disclosure provides a method of inhibiting the progression of dystrophic pathology associated with a muscular dystrophy, such as DMD. In some embodiments, the method comprises the step of administering one or more exon 17 U7-based snRNA polynucleotide construct without the use of a vector. In some embodiments, the method comprises the step of administering an rAAV to the patient, wherein the genome of the rAAV comprises an exon 17-targeted U7snRNA polynucleotide construct.

[0120] Combination therapies are also contemplated by the disclosure. Combination as used herein includes simultaneous treatment or sequential treatments. Combinations of methods of the disclosure with standard medical treatments (e.g., corticosteroids and / or immunosuppressive drugs) are specifically contemplated, as are combinations with other therapies such as those disclosed in International Publication No. WO 2013 / 016352, which is incorporated by reference herein in its entirety.

[0121] Each publication, patent application, patent, and other reference cited herein is incorporated by reference in its entirety to the extent that it is not inconsistent with the present disclosure.

[0122] Recitation of ranges of values herein are merely intended to serve as a shorthand method for referring individually to each separate value falling within the range and each endpoint, unless otherwise indicated herein, and each separate value and endpoint is incorporated into the specification as if it were individually recited herein.

[0123] All methods described herein are performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., "such as") provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0124] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims.EXAMPLES

[0125] Additional aspects and details of the disclosure will be apparent from the following examples, which are intended to be illustrative rather than limiting.Example 1Materials and Methods used in the Experiments

[0126] Cell lines

[0127] Exon skipping studies were performed in human cell lines carrying single exon 17 skip-amenable mutations (deletions of exons 18-33 (Dell 8-33) and 18-41 (Dell 8-41 )). These cell lines are a product of the Cell Line and Serum Banking Core, the shared resource core at Nationwide Children’s Hospital. Skin biopsies were obtained via a 3 mm punch biopsy under an existing approved protocol (IRB14-000719, “Evaluation of muscle, skin, and other tissues for the study of neuromuscular and neurologic diseases”). Primary fibroblast cultures were established in the Cell Line and Serum Banking core. Following immortalization with a lentiviral hTER gene, cells were infected with a tetracycline-inducible MyoD construct (tet-MyoD), after which cell culture in the presence of doxycycline induces transdifferentiation into the myoblast lineage. These FibroMyoD cells can subsequently be differentiated into myotubes to study dystrophin expression (Wein et al., J Neuromuscul Dis. 2017; 4(3):199-207; PMID28869484). All cells were grown in a humidified incubator at 37°C with 5% CO2.

[0128] Myoblast induction, myotube differentiation and AA V infection

[0129] FibroMyoD cells were plated into a 6-well plate and cultured using proliferation medium (DMEM 4.5 g / L glucose (Gibco), 20% fetal bovine serum (FBS; PAA Laboratories GmbH, Pasching, Austria), 100 mg / mL penicillin, 100 mg / mL streptomycin (Biowest, Nuaille, France), and L-glutamine (Gibco, Thermo Fisher Scientific, Waltham, MA, USA)). When cells reached approximately 85-90%confluence, the proliferation medium was switched to Skeletal Muscle Cell Growth Medium (SMGM, Promocell, C-23060, Heidelberg, Germany) with the addition of doxycycline (8 pig / ml; Sigma) to induce differentiation to myoblasts. At 95-100% confluency (2-3 days later), the SMGM medium was changed to Skeletal Muscle DifferentiationMedium (Promocell, C-23061 , Heidelberg, Germany) with doxycycline 8 .g / ml, in which cultures were maintained for 10-14 days. To promote differentiation, CHIR (5p.M; Axon Medchem, 2435) and DAPT (10p.M; Tocris, 2634) could also be added during that stage. rAAV1 injection was performed at the Myoblast stage, at 95-100% confluence and prior to myotube differentiation (Myoblast stage Day 2-3). FibroMyoD cell lines were treated with AAV1 vectors at the doses of 1 E2 to 1 E6 vg per cell. After 10-14 days of cell differentiation, cells were collected by adding TRIzol Reagent (0.5 ml per well, Invitrogen, 15596018) and stored at -80°C until RNA was extracted.

[0130] The DMDDel18~41mouse model

[0131] The DMDDel18-41mouse model was generated in the Center for Gene Therapy (Dr. Kevin Flanigan’s laboratory) at Nationwide Children’s Hospital by crossbreeding Dup2Del18-41 , which arose spontaneously on the background of the Dup2 mouse. As a result, this Dup2Del18- 41 mouse carries two frame-shift mutations. In order to create a mouse model carrying only exons 18-41 deletion, Dup2Del18-41mice were crossbred with Cre mouse model (B6.FVB-Tg(Ella- cre)C5379Lmgd / J, strain no. 003724) to remove one copy of duplicated exon 2.

[0132] DMDDenS 41intramuscular (IM) injections

[0133] For the IM injection studies, 12-week-old DMDDel18-41mice (n = 1 , 4 tissues) were injected in left and right sides of to the tibialis anterior (TA) and gastrocnemius (Gast) tissues at dose of 5E1 1 vg per muscle and euthanized 4 weeks later. TA and Gast tissues were collected from both legs and frozen in liquid-nitrogen-cooled isopentane. The DMD08118'41mouse model was generated in the Center for gene Therapy, Nationwide Children’s Hospital . This mouse model is used in various experiments described herein.

[0134] RNA extraction

[0135] RNA was extracted from human FM cell lines and DMD0811841mouse frozen tissues through TRIzol / chloroform extraction (Life Technologies, Carlsbad, CA, catalog no. 15596018; Fisher Bioreagents, Hampton, NH, catalog no. C297-4) and then purified using the RNA Clean & Concentrator-25 kit according to the manufacturer’s instruction (Zymo Research, Tustin, CA, catalog no. R1018).

[0136] Reverse transcription and PCR amplification

[0137] The RNA (1000 ng) was converted to cDNA using the RevertAid Reverse transcription kit and random hexamer primers according to the manufacturer’s protocol (ThermoScientific, Waltham, MA, catalog no. K1691 ). cDNA was amplified via PCR (Thermo Fisher, Waltham, MA, catalog no. K0171) using various sets of primers (Table 3) (IDT, Coralville, IA). The amplicons were visualized by gel electrophoresis and images were captured using the ChemiDoc MP imaging system (BioRad, Hercules, CA). Digital images of gels were quantified using Imaged software (version 1 ,46r, NIH, Bethesda, MD) to determine the relative amounts of different amplicons.

[0138] Table 3. RT-PCR primer sequences used in the study.

[0139] PCR amplification with an annealing temperature of 55-58sC was performed using the AccuStart™ II GelTrack PCR SuperMix (VWR, 89235-014).

[0140] Protein extraction and Western blotting

[0141] Protein was extracted using a RIPA lysis buffer (Cell signaling, catalog no. 9806S), and protein inhibitor cocktail (Sigma Aldrich, St. Louis, MO, catalog no. 11836170001 ). Briefly, 100-150 pL of lysis buffer was added to 10 sections of 20-mm thick tissue. Tissue was lysed using a metal bead and disrupted for 2 min at 30 Hz (TissueLyser II, Qiagen) followed by a 30- min incubation at room temperature before a second lysis step (1 min at 30 Hz, TissueLyser II, Qiagen). The lysate was centrifuged at 14,000 g for 20 min and the supernatant was collected for analysis. The protein concentration was quantified using the BioRad DC assay kit (BioRad, catalog no. 50001 12) following the manufacturer’s protocol. A calibration curve was made by combining lysates from BI6 mice (n = 3) and dystrophin-null age-matched Dup2Del18-41 mice (n = 3). These Dup2Del18-41 mice were received due to spontaneous mutation in the Dup2 colony and showed a complete absence of dystrophin protein. The supernatant was mixed with a 4 Laemmli sample buffer and heated for 5 min at 95°C. Total protein at 22.5 mg was run on aprecast 3%-8% Tris-Acetate NuPage gel (Invitrogen, catalog no. EA0378BOX) for 1 h at 80 V followed by 2 h at 120 V. Protein was transferred from gels to a 0.45-mm polyvinylidene fluoride membrane (BioRad, catalog no. 1620260) at a constant 55-mA current overnight at 4°C. Precision Plus Protein Dual Color Standards (BioRad, catalog no. 1610394) were used to determine the size of proteins of interest during separation and transfer. Membranes were probed with a rabbit monoclonal anti-dystrophin C-terminal antibody (Abeam, catalog no. ab154168) at 1 :1000 dilution in 5% nonfat dry milk in PBST buffer for 2 h. Blots were then washed 5 times each for 5 min (5 x 5) with PBST. Membranes were exposed to the secondary antibody, goat anti-rabbit horseradish peroxidase (1 :5000) for 1 h at room temperature, followed by 5 x 5-min washes with PBST and a 1 x 5-min wash with PBS. Membranes were incubated with 2 mL of enhanced chemiluminescence reagent (Thermo Scientific, catalog no. 34580) before visualization on a Chemidoc MP Imaging System (BioRad). Dystrophin signals were quantified by densitometric analysis using Image Lab software (BioRad, version 6.0.0, build 25). A linear regression curve fit to the calibration curve on each gel was calculated using the 0%- 50% WT dystrophin points, and individual samples were quantified against that curve and presented as percentage of dystrophin expression.

[0142] Membranes were probed with a rabbit monoclonal anti-dystrophin C-terminal antibody (Abeam, catalog no. ab154168) at 1 :1000 dilution in 5% nonfat dry milk in PBST buffer for 2 h. Blots were then washed five times for five minutes each with PBST. Membranes were exposed to the secondary antibody, goat anti-rabbit horseradish peroxidase (1 :5000) for 1 h at room temperature, followed by 5 x 5-min washes with PBST and a 1 x 5-min wash with PBS. Membranes were incubated with 2 mL of enhanced chemiluminescence reagent (Thermo Scientific, catalog no. 34580) before visualization on a Chemidoc MP Imaging System (BioRad). Dystrophin signals were quantified by densitometric analysis using Image Lab software (BioRad, version 6.0.0, build 25). A linear regression curve fit to the calibration curve on each gel was calculated using the 0%-50% WT dystrophin points, and individual samples were quantified against that curve and presented as percentage of dystrophin expression.Example 2Design and Generation of Sequences that Target Exon 17

[0143] In order to test the ability of the U7snRNA system to induce skipping of exon 17, five antisense sequences were designed. The sequences were designed using the Human Splicing Finder 3.0 program (Genomnics Inc.) (https_colon_forward slash forwardslash_bio.tools / human_splicing_finder) . Various target sequences and various targeting sequences with varying lengths and various binding sites were designed. Five antisense encoding sequences (i.e., SEQ ID NOs: 7, 13, 19, 25, and 31 ) were designed to bind or target exon 17 (one splice acceptor site (SAS) (SEQ ID NO: 7), one splice donor site (SDS) (SEQ ID NO: 31 ), and three exonic splicing enhancer (ESE) sites (SEQ ID NOs: 13, 19, and 25) in order to exclude an exon (e.g., exon 17) from the mRNA. Sequences were commercially synthesized (GenScript). These five antisense sequences were then used to make sequences encoding U7snRNA by placing them under the control of a U7 promoter or inserted into a sequence encoding U7 small nuclear RNA (U7 snRNA), thus arriving at the nucleotide sequences of SEQ ID NOs: 10, 16, 22, 28, and 34 which are shown in Table 2.

[0144] Tables 1 and 2 provide the sequences (nucleotide and amino acids) of exon 17 of the D / WD gene (and intronic sequence surrounding exon 17), target sequences on the DMD gene (exon 17 sequence (in upper case letters in SEQ ID NO: 1) and intronic sequence surrounding exon 17 (in lower case letters in SEQ ID NO: 1 ), antisense sequences used to target the sequences on the DMD gene (exon 17 and intronic sequence surrounding exon 17), reverse complement of the antisense sequences used to target the sequences on the DMD gene (exon 17 and intronic sequence surrounding exon 17), 117 sequences comprising antisense sequences used to target the sequences on the DMD gene (exon 17 and intronic sequence surrounding exon 17), and reverse complement of the U7 sequences comprising antisense sequences used to target the sequences on the DMD gene (exon 17 and intronic sequence surrounding exon 17).Example 3Development of Optimized rAAV.U7snRNA Constructs to Induce DMD Exon 17 Exclusion in Patient-Derived Cell Lines

[0145] As set forth above in Example 1 , five antisense sequences (i.e., SEQ ID NOs: 7, 13, 19, 25, and 31 ) were designed to test the ability of the U7snRNA system to induce skipping of exon 17. First generation U7snRNA constructs were then made, each construct comprising one copy of each of the splice acceptor site (SAS), splice donor site (SDS), or exon splicing enhancer site (ESE) antisense encoding sequences (i.e., SEQ ID NOs: 7, 13, 19, 25, and 31 ) were then inserted into a U7 expression construct comprising a U7 promoter to arrive at the U7snRNA sequences set forth in SEQ ID NOs: 10, 16, 22, 28, and 37.

[0146] Based on previous experience, exon skipping efficiency of U7snRNA constructs on the target site is enhanced or improved by adding multiple copies of the U7snRNA sequence. Thus, second generation U7snRNA constructs, containing either two copies (2xSDS, 2xSAS, or 2xESE) or four copies (4xSDS, 4xSAS, or 4xESE) of the U7snRNA sequences of the disclosure were made. See Table 4.

[0147] Table 4. Sequences of the U7snRNA constructs comprising multiple copies of the five RNA-encoding sense or antisense sequences of the disclosure.

[0148] The following U7snRNA constructs (nucleic acids comprising the nucleotide sequences of SEQ ID NOs: 42-61 ) containing multiple copies of the sense or antisense nucleotides encoding RNAs of the disclosure were designed to improve exon skipping efficiency.

[0149] 2x17SAS sense (874 bp) (SEQ ID NO: 42)

[0150] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATT TGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAA TATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGT GGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0151] 2x17SAS antisense (874 bp) (SEQ ID NO: 43)

[0152] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0153] 4x17SAS sense (1760 bp) (SEQ ID NO: 44)

[0154] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGgctagcgcatgcTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGaagcttagtactTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0155] 4x17SAS antisense (1760 bp) (SEQ ID NO: 45)

[0156] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAagtactaagcttCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAgcatgcgctagcCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0157] 2x17ESE1 sense (870 bp) (SEQ ID NO: 46)

[0158] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0159] 2x17ESE1 antisense (870 bp) (SEQ ID NO: 47)

[0160] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0161] 4x17ESE1 sense (1752 bp) (SEQ ID NO: 48)

[0162] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGgctagcgcatgcTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGaagcttagtactTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0163] 4x17ESE1 antisense (1752 bp) (SEQ ID NO: 49)

[0164] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACA- 61 -GTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAagtactaagcttCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAgcatgcgctagcCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0165] 2x17ESE2 sense (872 bp) (SEQ ID NO: 50)

[0166] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0167] 2x17ESE2 antisense (872 bp) (SEQ ID NO: 51 )

[0168] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0169] 4x17ESE2 sense (1756 bp) (SEQ ID NO: 52)

[0170] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGgctagcgcatgcTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGaagcttagtactTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0171] 4x17ESE2 antisense (1756 bp) (SEQ ID NO: 53)

[0172] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAagtactaagcttCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAgcatgcgctagcCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0173] 2x17ESE3 sense (866 bp) (SEQ ID NO: 54)

[0174] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0175] 2x17ESE3 antisense (866 bp) (SEQ ID NO: 55)

[0176] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTACTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTACTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0177] 4x17ESE3 sense (1744 bp) (SEQ ID NO: 56)

[0178] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGgctagcgcatgcTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGaagcttagtactTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0179] 4x17ESE3 antisense (1744 bp) (SEQ ID NO: 57)

[0180] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTACTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTACTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAagtactaagcttCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTACTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAgcatgcgctagcCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTACTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0181] 2x17SDS sense (876 bp) (SEQ ID NO: 58)

[0182] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATCTCACCTTTTCCTAATTTCAGAATAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATCTCACCTTTTCCTAATTTCAGAATAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0183] 2x17SDS antisense (876 bp) (SEQ ID NO: 59)

[0184] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTATTCTGAAATTAGGAAAAGGTGAGATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTATTCTGAAATTAGGAAAAGGTGAGATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA

[0185] 4x17SDS sense (1764 bp) (SEQ ID NO: 60)

[0186] TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATCTCACCTTTTCCTAATTTCAGAATAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGgctagcgcatgcTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATCTCACCTTTTCCTAATTTCAGAATAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGaagcttagtactTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATCTCACCTTTTCCTAATTTCAGAATAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATCTCACCTTTTCCTAATTTCAGAATAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG

[0187] 4x17SDS antisense (1764 bp) (SEQ ID NO: 61 )

[0188] CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTATTCTGAAATTAGGAAAAGGTGAGATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAccatggggatccCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTATTCTGAAATTAGGAAAAGGTGAGATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAagtactaagcttCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTATTCTGAAATTAGGAAAAGGTGAGATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAgcatgcgctagcCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTATTCTGAAATTAGGAAAAGGTGAGATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA- 16 -

[0189] For the initial screening, three rAAV plasmids comprising 17SAS, 17ESE1 , and 17SDS were made and purified, and sent to the Viral Vector Core (VVC) at Nationwide Children’s Hospital for insertion into a recombinant adeno-associated virus (rAAV) vector (i.e., between the ITRS) and three AAV1 ,U7.exon17 vectors were produced (Fig. 1 ). For the in vitro transduction studies, the constructs were produced using an AAV1 capsid. For in vivo studies, the constructs are produced into any AAV capsids as described herein. All vectors were produced using an AAV1 capsid at Andelyn Biosciences, formerly the NCH Viral Vector Core. All constructs (both generations 1 and 2) were tested via rAAV transduction in patient-derived cells for exon 17 skipping and dystrophin restoration. Two patient-derived FibroMyoD (FM) cell lines, carrying an out-of-frame deletion of exons 18-33 (Dell 8-33) and 18-41 (Dell 8-41 ) and suitable for exon 17 skipping, were generated from primary human fibroblasts (Wein et al., (2017). J Neuromuscul Dis 4, 199-207. 10.3233 / JND-170233). The advantage of the FM cells is that they can be transdifferentiated into a myoblast lineage, which can subsequently be differentiated into myotubes to study dystrophin expression (Wein et al., supra; Almeida et al., (2021 ). J Vis Exp. 10.3791 / 61991 ).

[0190] To detect exon 17 skipping, both FM cell lines were transdifferentiated into myoblasts, infected with four doses of AAV1 (from 1 E3 to 1 E6 vg / cell), differentiated into myotubes for 10-14 days, and then collected for RNA extraction. RT-PCR analysis detected the highest levels of exon 17 exclusion in both FM cell lines treated at the dose of 1 E6 vg / cell (Fig. 2A-B).

[0191] All constructs (both generations 1 and 2) were tested via rAAV transduction in patient-derived cells for exon 17 skipping and dystrophin restoration, and what appeared to be the three most efficacious candidates, i.e., 17SAS, 17ESE1 , and 17SDS, were selected for in vivo studies. Based on preliminary data, initial in vivo studies were carried out using these three constructs, including constructs comprising multiple copies of these constructs. Thus, in some particular experiments, constructs comprise the nucleotide sequences of SEQ ID NOs: 42-61 .

[0192] In some experiments, two copies or four copies of each of the sequences is used. Each sense / antisense sequence is driven by a U7 promoter. Thus, for example, when two copies are used, U7 / AS-U7 / AS are connected in series or U7 / S-U7 / S are connected in series, and, when four copies are used, U7 / AS-U7 / AS-U7 / AS-U7 / AS are connected in series or U7 / S- U7 / S-LI7 / S-U7 / S are connected in series. Therefore, these constructs are provided in either an antisense or sense orientation. In some embodiments, these constructs comprise the nucleotide sequences of SEQ ID NOs: 42-61 . These nucleotide sequences, provided in anantisense or sense orientation, are also referred to herein interchangeably as antisense or sense constructs. In some embodiments, these sequences or constructs are inserted into a vector for delivery into cells or into a subject.Example 4Initial In Vivo Testing of rAAV.U7snRNA Constructs to Induce DMD Exon 17 Exclusion in Mouse Model of DMD

[0193] For the initial in vivo studies, a study was designed as set out in Table 3 below.

[0194] Table 5. Initial in vivo study design.

[0195] DMDDel18-41mice (or controls (C57 / BI / 6)) were intramuscularly (IM) injected mice with three rAAV1 ,U7.exon17 vectors at the dose of 5E1 1 vg per leg. DMDDel18 41is a novel DMD mouse model carrying a deletion of exons 18-41 , which is the same mutation found in one of the two patient-derived cell lines that were studied. This mouse model was generated in the Center for Gene Therapy (Dr. Kevin Flanigan laboratory) at Nationwide Children’s Hospital.

[0196] Preliminary data revealed that to date, the 17SAS and 17SDS U7snRNA constructs were more effective in skipping / excluding exon 17 than the ESE U7snRNA construct (see Fig. 2A-E). Treatment with these U7.exon17 vectors comprising the 17SAS and 17SDS U7snRNAs resulted in the detection of in-frame DMD / dystrophin mRNA transcript by RT-PCR (Fig. 2C), and significant levels of dystrophin expression (up to 7.5% by western blotting (WB)) in mice 4 weeks post injection with the vectors (Fig. 2D-E).

[0197] These preliminary data demonstrate that IM injection of rAAV1 ,U7exon17 vectors with one copy of U7 in 12-week-old DMDDel18-41males at a dose of 5E1 1 vg / kg resulted in an average of 1 .8-1.9% in-frame Dell 7-41 DMD mRNA transcript in tissues from mice treated withrAAV1 .117.17SAS and rAAV1.U7.17SDS vectors, and up to 5.2% dystrophin expression in tissues after exonl 7SAS treatment (Fig. 2C-E). These data demonstrate that the exon 17 skipping strategy described herein is effective in obtaining dystrophin expression.Example 5In Vivo Testing of rAAV.U7snRNA Constructs to Induce DMD Exon 17 Exclusion in Mouse Model of DMD

[0198] To determine the effectiveness of the new generation of vectors and whether they can improve muscle pathology and muscle function in dystrophic mice, the three bestperforming candidates are packaged into a new vector, rMYOAAV3A, and administered intravenously in 1 1-13-week-old DMDDel18-41males at a dose of 1 E13 vg / kg (n=6) (Table 4).

[0199] Table 6. rMYOAAV3A in vivo study design.

[0200] This dose was chosen based on previous exon 2 skipping studies in Dup2 mice (Simmons et al., Mol Ther Methods Clin Dei / 21 , 325-340 (2021 ); 10.1016 / j.omtm.2021.03.014) and estimated efficacy published for myotrophic serotypes (Tabebordbar et al. (2021 ) Cell 184, 4919-4938 e4922. 10.1016 / j . cell.2021 .08.028). Animals are euthanized 4 weeks post-vector injection. Before euthanasia, two electrophysiology tests are carried out as a functional readout of therapy efficacy: in situ absolute and specific tetanic force output (providing an assessment of strength), and eccentric contraction measures (evaluating sarcolemmal stability) in the tibialis anterior (TA) muscle, and in vitro maximal tetanic force in isolated muscle strips dissected from the diaphragm, as has been reported in previous studies (Gushchina et al. (2023). Mol Ther Methods Clin Dei / 31 , 101144. 10.1016 / j.omtm.2023.101144). Skeletal muscles (both left and right TA, gastroc, quadriceps, triceps, gluteus maximus, biceps, diaphragm and heart) are dissected from tendon to tendon, and then snap-frozen in LN2-cooled isopentane and stored at-80°C until use. Liver, spleen, kidney, lung, testis, brain and tail vein site of injection are also collected for biodistribution and pathology analysis. Serum is collected to measure serum creatine kinase activity and lactate dehydrogenase, biomarkers for muscle damage (Gushchina, et al. (2017) Mol Ther25, 2360-2371. 10.1016 / j.ymthe.2O17.06.025) and to assess AAV antibody titers (Chicoine et al. (2014). Mol Ther 22, 338-347. 10.1038 / mt.2013.244). Transgene biodistribution, exon 17 skipping efficacy, and dystrophin expression will be assessed by ddPCR, RT-PCR, capillary western, and immunofluorescence analysis, including the percentage of dystrophin-positive fibers (PDPFs) and dystrophin intensity (Gushchina et al. (2023) Mol Ther Methods Clin Dev 31 , 101144. 10.1016 / j.omtm.2023.101144; Vetter et al. (2022). Neuropathol Appl Neurobiol 48, e12785. 10.1 11 1 / nan.12785). All experiments are carried out in a blinded fashion.

[0201] Significantly greater exon 17 skipping and dystrophin rescue is obtained using 2ndgeneration vectors resulting in stable D / WD mRNA, leading to functional dystrophin expression, and a significant increase in muscle specific force in both TA and diaphragm muscles, as well as significant improvement of the force drop during repeated eccentric contractions in the TA muscle.Example 6Additional Constructs to Induce DMD Exon 17 Exclusion

[0202] In an effort to improve or increase efficacy of exon skipping of the snRNAs of the disclosure, additional first and generation constructs were made by adding stuffer(s), promoter(s), and / or enhancer(s), and / or additional copies of small nuclear RNAs in a construct. These constructs were designed to enhance the expression of a functional but altered form of dystrophin protein for treating a muscular dystrophy by inducing DMD exon 17 exclusion. The design of these additional constructs is exemplified in Fig. 1 ; however, specifically designed constructs are described herein in detail below in this Example and in Table 7.

[0203] Stuffer sequence was added to each of the five first generation constructs for each of 17SAS, 17ESE1 , 17ESE2, 17ESE3, and 17SDS. Those sequences are found in SEQ ID NOs: 62-71 in both the sense and antisense orientations. The plasmid sequence for each of these sequences is further found in SEQ ID NOs: 72-81 as provided in detail below.

[0204] Stuffer and enhancer sequences were added to each of the five second generation constructs for each of 17SAS, 17ESE1 , 17ESE2, 17ESE3, and 17SDS. Those sequences are found in SEQ ID NOs: 82-121 in both the sense and antisense orientations as provided in detailbelow. The plasmid sequence for each of these sequences is further found in SEQ ID NOs: 122-16 as provided in detail below. The design of these sequences is exemplified in Fig. 1 .

[0205] Table 7. Names and sequence identification numbers of the additional snRNA constructs of the disclosure. The sequences of these additional snRNA constructs are provided after the table in this example.

[0206] The sequences of the additional snRNA constructs of the disclosure (both sense and antisense) and their sequences identification numbers are provided as set out below.1stgeneration of constructs1. U7.17SAS with stuffer, sense sequence (1949 bp) SEQ ID NO: 62GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACGTCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCTAGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACATTCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAG AGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCC GCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGG GGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTT TAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATG GCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCT GTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAA AACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStaffer sense sequence 1X17SAS sense sequence2. U7.17SAS with stuffer, antisense sequence (1949 bp) SEQ ID NO: 63CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGC TGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAGCTCCTAJGTrGTrACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCC ACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAAT TCCGCAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCA CCGCGGTGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTAC TGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTG GGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGAT GATGGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGA GCGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGA AAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCAC CCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCG CGAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTCGCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAG GCGGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGAT CCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCG CTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACC1X17SAS antisense sequenceStuffer antisense sequence3. U7.17ESE1 with stuffer, sense sequence (1947 bp) SEQ ID NO: 64GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTG CCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGC AGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTT GACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACG TCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAA GCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGA TCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAG GCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCG CGCCGGGGGGGGGGGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGC CAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGG TGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCT AGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACAT TCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAG AGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCC GCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGG GGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACMAGGAGCT GTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTT TAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATG GCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAA CAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStaffer sense sequence 1X17ESE1 sense sequence4. U7.17ESE1 with stuffer, antisense sequence (1947 bp) SEQ ID NO: 65CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACAC AGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCAT TTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAAC CGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTAJGTrGnACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAC TGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTC CGCAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACC GCGGTGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACT GGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGG GCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCC AGGCTTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATG ATGGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAG CGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAA GGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCC TAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTT CTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGCG AAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTCGCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAGGC GGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTG GCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGATCCT TGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCGCTT GCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACC1X17ESE1 antisense sequenceStuffer antisense sequence5. U7.17ESE2 with stuffer, sense sequence (1948 bp) SEQ ID NO: 66GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTG CCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTT GACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACG TCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAA GCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTT TTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGA TCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAA GAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAG GCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCG CGCCGGGGGGGGGGGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGC CAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGG TGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCT AGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACAT TCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAG AGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGG GGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTT TAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATG GCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTT CCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAA ACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStaffer sense sequence 1X17ESE2 sense sequence6. U7.17ESE2 with stuffer, antisense sequence (1948 bp) SEQ ID NO: 67CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACA GATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAG CTCCTMGTTGTTf^CATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAAT TCCGCAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCA CCGCGGTGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTAC TGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTG GGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCC CAGGCTTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGAT GATGGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGA GCGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGA AAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCAC CCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAAT TTCTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGCGAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTC GCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAG GCGGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATC TGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGAT CCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCG CTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACC1X17ESE2 antisense sequence Stuffer antisense sequence7. U7.17ESE3 with stuffer, sense sequence (1945 bp) SEQ ID NO: 68GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTG CCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGC AGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTT GACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACG TCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAA GCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTT TTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGA TCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAA GAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAG GCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCG CGCCGGGGGGGGGGGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGG TGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCT AGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACAT TCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAG AGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCC GCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGG GGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCT GTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTT TAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATG GCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTG CCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer sense sequence 1X17ESE3 sense sequence8. U7.17ESE3 with stuffer, antisense sequence (1945 bp) SEQ ID NO: 69CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTA CTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTT CCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCG CTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCC MGnGJJACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTG ATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCG CAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACCGC GGTGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGG GCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGC GGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAG GCTTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGAT GGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGC GGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAA GGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCC TAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGCG AAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTCGC GCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAGGC GGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTG GCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGATCCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCGCTT GCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACC1X17ESE3 antisense sequenceStaffer antisense sequence9. U7.17SDS with stuffer, sense sequence (1950 bp) SEQ ID NO: 70GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTG CCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGC AGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTT GACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACG TCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTT TTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGA TCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAA GAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCG CGCCGGGGGGGGGGGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGC CAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGG TGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCT AGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACATTCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAG AGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCC GCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGG GGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTT TAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATG GCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATCTCACCTTTTCC TAATTTCAGAATAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAA AACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer sense sequence 1X17SDS sense sequence10. U7.17SDS with stuffer, antisense sequence (1950 bp) SEQ ID NO: 71CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTATTCTGAAATTAGGAA AAGGTGAGATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTG CCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTA AAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTMGJIG IACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCC CCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACA ATTCCGCAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGC CACCGCGGTGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCT ACTGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACC CCAGGCTTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGA TGATGGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTG AGCGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATG AAAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCA CCCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAA TTTCTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCC GCGAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGC TCGCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCA GGCGGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATAT CTGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGA TCCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTC GCTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACC1X17SDS antisense sequence Staffer antisense sequencePlasmid sequences:1. AAV.U7.17SAS, sense plasmid sequence (5922 bp) SEQ ID NO: 72CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGAATTCACGCGTGGATCTGAATTCAATTCACGCGTGGTACCCTTAAGTCTAGAGGCTCGAG AAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGG TTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGCTCTGATCAAGAGA CAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAA TAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACGTCGACGGATCGGGAGATCTCCCGA TCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGAC CGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTT AAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTT CTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCC GCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAAT CTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCG CTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCC ACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTA ACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCTAGCTATCGCCATGTAAG CCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACATTCATCCGGGGTCAGC ACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAGAGTCGACGAGGAA CTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCAC CGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGG ACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGT TTTCAG CCA ATCAG CACTG ACTCATTTG CATAG CCTTTACA AG CG GTC ACA AACTC AAG AAACG AG CG GTTTTAATAGTCTTTTAG AATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCAC CCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATT TTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGC TCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGCTAGACTAGAGCATGGCTACGTAGA TAAGTAGCATGGCGGGTTAATCATTAACTACAAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCA CTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGG GAGTGGCCCCCCCCCCCCCCCCCCCGGCGATTCTCTTGTTTGCTCCAGACTCTCAGGCAATGACCTGATAGCCTTTGTAGAGACCT CTCAAAAATAGCTACCCTCTCCGGCATGAATTTATCAGCTAGAACGGTTGAATATCATATTGATGGTGATTTGACTGTCTCCGGCCTTTCTCACCCGTTTGAATCTTTACCTACACATTACTCAGGCATTGCATTTAAAATATATGAGGGTTCTAAAAATTTTTATCCTTGCGTTG AAATA AAG G CTTCTCCCG C AAAAGTATTAC AG G GTCATAATGTTTTTG GTACAACCG ATTTAG CTTTATG CTCTG AG G CTTTATTG C TTAATTTTGCTAATTCTTTGCCTTGCCTGTATGATTTATTGGATGTTGGAATCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACTAT GGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACG GGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCA CCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGG TGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAA CCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGCCATATTCAACGGGAAACGTCGAGGCCGCGATTAAATTCCAA CATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGCTTGTATGGGAAGC CCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCT GACGGAATTTATGCCACTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGAAA AACAGCGTTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCACT CGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGCCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTG ATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACC GGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACG AGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTT TTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAACTGTCAGACCAAGT TTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCA AAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTG CGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTC CGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTC TGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGG ACTCA AG ACG ATAGTTACCG G ATA AGG CGCAG CG GTCG G G CTG AACG G G GG GTTCGTGC AC AC AG CCCAG CTTG G AG CG AAC GACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATC CGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGG GTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGC CTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACC GCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCC CAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCC TTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGATTCCGTTGCAATGGCTGGCGGTAATATTGTTCTGGATATTACCAGCAAG GCCGATAGTTTGAGTTCTTCTACTCAGGCAAGTGATGTTATTACTAATCAAAGAAGTATTGCGACAACGGTTAATTTGCGTGATGG ACAGACTCTTTTACTCGGTGGCCTCACTGATTATAAAAACACTTCTCAGGATTCTGGCGTACCGTTCCTGTCTAAAATCCCTTTAATC GGCCTCCTGTTTAGCTCCCGCTCTGATTCTAACGAGGAAAGCACGTTATACGTGCTCGTCAAAGCAACCATAGTACGCGCCCTGTA GCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCG CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTG CTTTACG G CACCTCG ACCCCAAAAAACTTG ATTAG G GTG ATG GTTCACGTAGTG G G CCATCG CCCTG ATAG ACG GTTTTTCG CCC TTTG ACGTTG G AGTCC ACGTTCTTTAATAGTG G ACTCTTGTTCCA AACTG G AAC AACACTC AACCCTATCTCG GTCTATTCTTTTG A TTTATAAG G G ATTTTG CCG ATTTCG G CCTATTG GTTAAAAAATG AGCTG ATTTAACAAA AATTTA ACG CG AATTTTA AC AAAATATT AACGCTTACAATTTAAATATTTGCTTATACAATCTTCCTGTTTTTGGGGCTTTTCTGATTATCAACCGGGGTACATATGATTGACATGC TAGTTTTACGATTACCGTTCATCGCC2. AAV.U7.17SAS, antisense plasmid sequence (5922 bp) SEQ ID NO: 73GGCGATGAACGGTAATCGTAAAACTAGCATGTCAATCATATGTACCCCGGTTGATAATCAGAAAAGCCCCAAAAACAGGAAGATT GTATAAGCAAATATTTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATA GGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCC ACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAA TCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAA GCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGC TGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTACTATGGTTGCTTTGACGAGCACGTATAACGTGC TTTCCTCGTTAGAATCAGAGCGGGAGCTAAACAGGAGGCCGATTAAAGGGATTTTAGACAGGAACGGTACGCCAGAATCCTGAG AAGTGTTTTTATAATCAGTGAGGCCACCGAGTAAAAGAGTCTGTCCATCACGCAAATTAACCGTTGTCGCAATACTTCTTTGATTAG TAATAACATCACTTGCCTGAGTAGAAGAACTCAAACTATCGGCCTTGCTGGTAATATCCAGAACAATATTACCGCCAGCCATTGCAA CGGAATCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGCATT AATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGG TCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCT GACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGA AGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTC TCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCC GACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTA ACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAG TATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGG TAG CG GTG GTTTTTTTGTTTG CAAG CAG C AG ATTACG CG CAG AAAAAA AG G ATCTCA AG A AG ATCCTTTG ATCTTTTCTACG G G G TCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAA TTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTAGAAAAACTCATCGAGCATCAAATGAAA CTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTT CCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAAT AAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCAGACTTGT TCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGGCG AAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAGTGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAAT ATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAACGCTGTTTTTCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGG AGTACG G ATA AAATG CTTG ATG GTCG G AAGTG G CATAAATTCCGTCAG CC AGTTTAGTCTG ACCATCTCATCTGTAACATC ATTG G C AACG CTACCTTTG CCATGTTTCAG AAACAACTCTG GCG CATCG G G CTTCCC ATACA AG CG ATAG ATTGTCG CACCTG ATTG CCCG A CATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTCGACGTTTCCCGTTGAATATGGCT CATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATA AACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAA AATAGGCGTATCACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACG GTCACAG CTTGTCTGTAAG CG G ATG CCG GG AG CAG ACAAG CCCGTCAG G G CG CGTC AGCG G GTGTTG G CG G GTGTCG G G G CT GGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATAGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGCGG CATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGC G ATTCCAAC ATCCA ATAA ATCATACAGG CAAG G CAAAG AATTAG CAAAATTAAG CAATAA AG CCTCAG AG CATAA AG CTAAATCG GTTGTACCAAAAACATTATGACCCTGTAATACTTTTGCGGGAGAAGCCTTTATTTCAACGCAAGGATAAAAATTTTTAGAACCCTCA TATATTTTAAATGCAATGCCTGAGTAATGTGTAGGTAAAGATTCAAACGGGTGAGAAAGGCCGGAGACAGTCAAATCACCATCAAT ATGATATTCAACCGTTCTAGCTGATAAATTCATGCCGGAGAGGGTAGCTATTTTTGAGAGGTCTCTACAAAGGCTATCAGGTCATTG CCTGAGAGTCTGGAGCAAACAAGAGAATCGCCGGGGGGGGGGGGGGGGGGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCAC TGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGG AGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTAGCCATGCTCTAGTCTAGCC ACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTC ATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTC ACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCC ACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTC GTTTCTTG AGTTTGTG ACCG CTTGTAA AG G CTATG CAAATG AGTCAGTG CTG ATTG G CTG AAAACAG CC AATCACAG CTCCTATGT TGTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATCA GCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCGCAG CTTTTAG AG CAG AAGTAACACTTCCGTACAGG CCAGTTAACTTTCTG GTTTTTC AGTTCCTCGTCG ACTCTAG GCCGCCACCGCGG TGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGC TATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGG GCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGC TTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATG GGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCG GGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAG GTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCT AGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTT CTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGC GAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTC GCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAG GCGGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGAT CCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTC GCTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACCACGCGTGAATTGA ATTCAGATCCACGCGTGAATTCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCC GGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAG3. AAV.U7.17ESE1, sense plasmid sequence (5920 bp) SEQ ID NO: 74CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGAATTCACGCGTGGATCTGAATTCAATTCACGCGTGGTACCCTTAAGTCTAGAGGCTCGAG AAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGG TTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGCTCTGATCAAGAGA CAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAA TAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACGTCGACGGATCGGGAGATCTCCCGA TCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGAC CGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTT AAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTT CTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCC GCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAAT CTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCG CTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCC ACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTA ACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCTAGCTATCGCCATGTAAG CCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACATTCATCCGGGGTCAGC ACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAGAGTCGACGAGGAA CTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCAC CGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGG ACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGT TTTCAG CCA ATCAG CACTG ACTCATTTG CATAG CCTTTACA AG CG GTC ACA AACTC AAG AAACG AG CG GTTTTAATAGTCTTTTAG AATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCAC CCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTT TGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTC CCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGCTAGACTAGAGCATGGCTACGTAGATA AGTAGCATGGCGGGTTAATCATTAACTACAAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACT GAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGA GTGGCCCCCCCCCCCCCCCCCCCGGCGATTCTCTTGTTTGCTCCAGACTCTCAGGCAATGACCTGATAGCCTTTGTAGAGACCTCT CAAAAATAGCTACCCTCTCCGGCATGAATTTATCAGCTAGAACGGTTGAATATCATATTGATGGTGATTTGACTGTCTCCGGCCTTT CTCACCCGTTTGAATCTTTACCTACACATTACTCAGGCATTGCATTTAAAATATATGAGGGTTCTAAAAATTTTTATCCTTGCGTTGA AATAA AG G CTTCTCCCG CA AAAGTATTACAG G GTC ATA ATGTTTTTG GTACAACCG ATTTAG CTTTATG CTCTG AG G CTTTATTG CTTAATTTTGCTAATTCTTTGCCTTGCCTGTATGATTTATTGGATGTTGGAATCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGG TATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACTATG GTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGG GCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCAC CGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGT GGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAA CCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGCCATATTCAACGGGAAACGTCGAGGCCGCGATTAAATTCCAA CATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGCTTGTATGGGAAGC CCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCT GACGGAATTTATGCCACTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGAAA AACAGCGTTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCACT CGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGCCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTG ATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACC GGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACG AGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAACTGTCAGACCAAGT TTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCA AAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTG CGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTC CGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTC TGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGG ACTCA AG ACG ATAGTTACCG G ATA AGG CGCAG CG GTCG G G CTG AACG G G GG GTTCGTGC AC AC AG CCCAG CTTG G AG CG AAC GACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATC CGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGG GTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGC CTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACC GCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCC CAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCC TTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGATTCCGTTGCAATGGCTGGCGGTAATATTGTTCTGGATATTACCAGCAAG GCCGATAGTTTGAGTTCTTCTACTCAGGCAAGTGATGTTATTACTAATCAAAGAAGTATTGCGACAACGGTTAATTTGCGTGATGG ACAGACTCTTTTACTCGGTGGCCTCACTGATTATAAAAACACTTCTCAGGATTCTGGCGTACCGTTCCTGTCTAAAATCCCTTTAATC GGCCTCCTGTTTAGCTCCCGCTCTGATTCTAACGAGGAAAGCACGTTATACGTGCTCGTCAAAGCAACCATAGTACGCGCCCTGTA GCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCG CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTG CTTTACG G CACCTCG ACCCCAAAAAACTTG ATTAG G GTG ATG GTTCACGTAGTG G G CCATCG CCCTG ATAG ACG GTTTTTCG CCC TTTG ACGTTG G AGTCC ACGTTCTTTAATAGTG G ACTCTTGTTCCA AACTG G AAC AACACTC AACCCTATCTCG GTCTATTCTTTTG A TTTATAAG G G ATTTTG CCG ATTTCG G CCTATTG GTTAAAAAATG AGCTG ATTTAACAAA AATTTA ACG CG AATTTTA AC AAAATATT AACGCTTACAATTTAAATATTTGCTTATACAATCTTCCTGTTTTTGGGGCTTTTCTGATTATCAACCGGGGTACATATGATTGACATGC TAGTTTTACGATTACCGTTCATCGCC4. AAV.U7.17ESE1, antisense plasmid sequence (5920 bp) SEQ ID NO: 75GGCGATGAACGGTAATCGTAAAACTAGCATGTCAATCATATGTACCCCGGTTGATAATCAGAAAAGCCCCAAAAACAGGAAGATT GTATAAGCAAATATTTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATA GGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCC ACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAA TCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAA GCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGC TGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTACTATGGTTGCTTTGACGAGCACGTATAACGTGC TTTCCTCGTTAGAATCAGAGCGGGAGCTAAACAGGAGGCCGATTAAAGGGATTTTAGACAGGAACGGTACGCCAGAATCCTGAG AAGTGTTTTTATAATCAGTGAGGCCACCGAGTAAAAGAGTCTGTCCATCACGCAAATTAACCGTTGTCGCAATACTTCTTTGATTAG TAATAACATCACTTGCCTGAGTAGAAGAACTCAAACTATCGGCCTTGCTGGTAATATCCAGAACAATATTACCGCCAGCCATTGCAA CGGAATCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGCATT AATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGG TCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGA ACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCT GACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGA AGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTC TCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCC GACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTA ACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAG TATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGG TAG CG GTG GTTTTTTTGTTTG CAAG CAG C AG ATTACG CG CAG AAAAAA AG G ATCTCA AG A AG ATCCTTTG ATCTTTTCTACG G G G TCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAA TTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTAGAAAAACTCATCGAGCATCAAATGAAA CTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTT CCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAAT AAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCAGACTTGT TCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGGCGAAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAGTGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAAT ATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAACGCTGTTTTTCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGG AGTACG G ATA AAATG CTTG ATG GTCG G AAGTG G CATAAATTCCGTCAG CC AGTTTAGTCTG ACCATCTCATCTGTAACATC ATTG G C AACG CTACCTTTG CCATGTTTCAG AAACAACTCTG GCG CATCG G G CTTCCC ATACA AG CG ATAG ATTGTCG CACCTG ATTG CCCG A CATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTCGACGTTTCCCGTTGAATATGGCT CATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATA AACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAA AATAGGCGTATCACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACG GTCACAG CTTGTCTGTAAG CG G ATG CCG GG AG CAG ACAAG CCCGTCAG G G CG CGTC AGCG G GTGTTG G CG G GTGTCG G G G CT GGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATAGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGCGG CATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGC G ATTCCAAC ATCCA ATAA ATCATACAGG CAAG G CAAAG AATTAG CAAAATTAAG CAATAA AG CCTCAG AG CATAA AG CTAAATCG GTTGTACCAAAAACATTATGACCCTGTAATACTTTTGCGGGAGAAGCCTTTATTTCAACGCAAGGATAAAAATTTTTAGAACCCTCA TATATTTTAAATGCAATGCCTGAGTAATGTGTAGGTAAAGATTCAAACGGGTGAGAAAGGCCGGAGACAGTCAAATCACCATCAAT ATGATATTCAACCGTTCTAGCTGATAAATTCATGCCGGAGAGGGTAGCTATTTTTGAGAGGTCTCTACAAAGGCTATCAGGTCATTG CCTGAGAGTCTGGAGCAAACAAGAGAATCGCCGGGGGGGGGGGGGGGGGGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCAC TGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGG AGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTAGCCATGCTCTAGTCTAGCC ACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTC ATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGAC AATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCAC ACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGT TTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTG TTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATCAGC GAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCGCAGCT TTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACCGCGGT GGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCT ATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGG GCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGC TTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATG GGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCG GGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAG GTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCT AGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTT CTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGC GAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTC GCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAG GCGGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATC TGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGAT CCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTC GCTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACCACGCGTGAATTGA ATTCAGATCCACGCGTGAATTCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCC GGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAG5. AAV.U7.17ESE2, sense plasmid sequence (5921 bp) SEQ ID NO: 76CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGAATTCACGCGTGGATCTGAATTCAATTCACGCGTGGTACCCTTAAGTCTAGAGGCTCGAG AAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGG TTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGCTCTGATCAAGAGA CAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAA TAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACGTCGACGGATCGGGAGATCTCCCGA TCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGAC CGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTT CTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCC GCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAAT CTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCG CTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCC ACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTA ACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCTAGCTATCGCCATGTAAG CCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACATTCATCCGGGGTCAGC ACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAGAGTCGACGAGGAA CTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCAC CGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGG ACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGT TTTCAG CCA ATCAG CACTG ACTCATTTG CATAG CCTTTACA AG CG GTC ACA AACTC AAG AAACG AG CG GTTTTAATAGTCTTTTAG AATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCAC CCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTT TTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCT CCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGCTAGACTAGAGCATGGCTACGTAGAT AAGTAGCATGGCGGGTTAATCATTAACTACAAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCA CTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGG GAGTGGCCCCCCCCCCCCCCCCCCCGGCGATTCTCTTGTTTGCTCCAGACTCTCAGGCAATGACCTGATAGCCTTTGTAGAGACCT CTCAAAAATAGCTACCCTCTCCGGCATGAATTTATCAGCTAGAACGGTTGAATATCATATTGATGGTGATTTGACTGTCTCCGGCCT TTCTCACCCGTTTGAATCTTTACCTACACATTACTCAGGCATTGCATTTAAAATATATGAGGGTTCTAAAAATTTTTATCCTTGCGTTG AAATA AAG G CTTCTCCCG C AAAAGTATTAC AG G GTCATAATGTTTTTG GTACAACCG ATTTAG CTTTATG CTCTG AG G CTTTATTG C TTAATTTTGCTAATTCTTTGCCTTGCCTGTATGATTTATTGGATGTTGGAATCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCG GTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACTAT GGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACG GGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCA CCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGG TGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAA CCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGCCATATTCAACGGGAAACGTCGAGGCCGCGATTAAATTCCAA CATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGCTTGTATGGGAAGC CCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCT GACGGAATTTATGCCACTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGAAA AACAGCGTTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCACT CGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGCCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTG ATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACC GGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACG AGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTT TTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAACTGTCAGACCAAGT TTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCA AAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTG CGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTC CGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTC TGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGG ACTCA AG ACG ATAGTTACCG G ATA AGG CGCAG CG GTCG G G CTG AACG G G GG GTTCGTGC AC AC AG CCCAG CTTG G AG CG AAC GACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGG GTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGC CTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACC GCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCC CAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCC TTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGATTCCGTTGCAATGGCTGGCGGTAATATTGTTCTGGATATTACCAGCAAGGCCGATAGTTTGAGTTCTTCTACTCAGGCAAGTGATGTTATTACTAATCAAAGAAGTATTGCGACAACGGTTAATTTGCGTGATGG ACAGACTCTTTTACTCGGTGGCCTCACTGATTATAAAAACACTTCTCAGGATTCTGGCGTACCGTTCCTGTCTAAAATCCCTTTAATC GGCCTCCTGTTTAGCTCCCGCTCTGATTCTAACGAGGAAAGCACGTTATACGTGCTCGTCAAAGCAACCATAGTACGCGCCCTGTA GCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCG CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTG CTTTACG G CACCTCG ACCCCAAAAAACTTG ATTAG G GTG ATG GTTCACGTAGTG G G CCATCG CCCTG ATAG ACG GTTTTTCG CCC TTTG ACGTTG G AGTCC ACGTTCTTTAATAGTG G ACTCTTGTTCCA AACTG G AAC AACACTC AACCCTATCTCG GTCTATTCTTTTG A TTTATAAG G G ATTTTG CCG ATTTCG G CCTATTG GTTAAAAAATG AGCTG ATTTAACAAA AATTTA ACG CG AATTTTA AC AAAATATT AACGCTTACAATTTAAATATTTGCTTATACAATCTTCCTGTTTTTGGGGCTTTTCTGATTATCAACCGGGGTACATATGATTGACATGC TAGTTTTACGATTACCGTTCATCGCC6. AAV.U7.17ESE2, antisense plasmid sequence (5921 bp) SEQ ID NO: 77GGCGATGAACGGTAATCGTAAAACTAGCATGTCAATCATATGTACCCCGGTTGATAATCAGAAAAGCCCCAAAAACAGGAAGATT GTATAAGCAAATATTTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATA GGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCC ACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAA TCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAA GCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGC TGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTACTATGGTTGCTTTGACGAGCACGTATAACGTGC TTTCCTCGTTAGAATCAGAGCGGGAGCTAAACAGGAGGCCGATTAAAGGGATTTTAGACAGGAACGGTACGCCAGAATCCTGAG AAGTGTTTTTATAATCAGTGAGGCCACCGAGTAAAAGAGTCTGTCCATCACGCAAATTAACCGTTGTCGCAATACTTCTTTGATTAG TAATAACATCACTTGCCTGAGTAGAAGAACTCAAACTATCGGCCTTGCTGGTAATATCCAGAACAATATTACCGCCAGCCATTGCAA CGGAATCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGCATT AATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGG TCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGA ACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCT GACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGA AGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTC TCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCC GACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTA ACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAG TATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGG TAG CG GTG GTTTTTTTGTTTG CAAG CAG C AG ATTACG CG CAG AAAAAA AG G ATCTCA AG A AG ATCCTTTG ATCTTTTCTACG G G G TCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAA TTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTAGAAAAACTCATCGAGCATCAAATGAAA CTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTT CCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAAT AAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCAGACTTGT TCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGGCG AAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAGTGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAAT ATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAACGCTGTTTTTCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGG AGTACG G ATA AAATG CTTG ATG GTCG G AAGTG G CATAAATTCCGTCAG CC AGTTTAGTCTG ACCATCTCATCTGTAACATC ATTG G C AACG CTACCTTTG CCATGTTTCAG AAACAACTCTG GCG CATCG G G CTTCCC ATACA AG CG ATAG ATTGTCG CACCTG ATTG CCCG A CATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTCGACGTTTCCCGTTGAATATGGCT CATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATA AACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAA AATAGGCGTATCACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACG GTCACAG CTTGTCTGTAAG CG G ATG CCG GG AG CAG ACAAG CCCGTCAG G G CG CGTC AGCG G GTGTTG G CG G GTGTCG G G G CT GGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATAGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGCGG CATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGC G ATTCCAAC ATCCA ATAA ATCATACAGG CAAG G CAAAG AATTAG CAAAATTAAG CAATAA AG CCTCAG AG CATAA AG CTAAATCGGTTGTACCAAAAACATTATGACCCTGTAATACTTTTGCGGGAGAAGCCTTTATTTCAACGCAAGGATAAAAATTTTTAGAACCCTCA TATATTTTAAATGCAATGCCTGAGTAATGTGTAGGTAAAGATTCAAACGGGTGAGAAAGGCCGGAGACAGTCAAATCACCATCAATATGATATTCAACCGTTCTAGCTGATAAATTCATGCCGGAGAGGGTAGCTATTTTTGAGAGGTCTCTACAAAGGCTATCAGGTCATTG CCTGAGAGTCTGGAGCAAACAAGAGAATCGCCGGGGGGGGGGGGGGGGGGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCAC TGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGG AGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTAGCCATGCTCTAGTCTAGCC ACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTC ATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCC TGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCA CACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCG TTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTT GTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATCAG CGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCGCAGC TTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACCGCGGT GGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCT ATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGG GCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGC TTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATG GGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCG GGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAG GTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCT AGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTT CTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGC GAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTC GCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAG GCGGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATC TGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGAT CCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCGCTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACCACGCGTGAATTGA ATTCAGATCCACGCGTGAATTCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCC GGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAG7. AAV.U7.17ESE3, sense plasmid sequence (5918 bp) SEQ ID NO: 78CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGAATTCACGCGTGGATCTGAATTCAATTCACGCGTGGTACCCTTAAGTCTAGAGGCTCGAG AAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGCTCTGATCAAGAGA CAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAA TAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACGTCGACGGATCGGGAGATCTCCCGA TCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGAC CGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTT AAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTT CTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCC GCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAAT CTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCG CTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCC ACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTA ACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCTAGCTATCGCCATGTAAG CCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACATTCATCCGGGGTCAGC ACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAGAGTCGACGAGGAA CTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCAC CGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGG ACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGT TTTCAG CCA ATCAG CACTG ACTCATTTG CATAG CCTTTACA AG CG GTC ACA AACTC AAG AAACG AG CG GTTTTAATAGTCTTTTAG AATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCAC- 91 -CCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTG GAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCC CGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGCTAGACTAGAGCATGGCTACGTAGATAAG TAGCATGGCGGGTTAATCATTAACTACAAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGA GGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT GGCCCCCCCCCCCCCCCCCCCGGCGATTCTCTTGTTTGCTCCAGACTCTCAGGCAATGACCTGATAGCCTTTGTAGAGACCTCTCA AAAATAGCTACCCTCTCCGGCATGAATTTATCAGCTAGAACGGTTGAATATCATATTGATGGTGATTTGACTGTCTCCGGCCTTTCTC ACCCGTTTGAATCTTTACCTACACATTACTCAGGCATTGCATTTAAAATATATGAGGGTTCTAAAAATTTTTATCCTTGCGTTGAAAT AAAGGCTTCTCCCGCAAAAGTATTACAGGGTCATAATGTTTTTGGTACAACCGATTTAGCTTTATGCTCTGAGGCTTTATTGCTTAA TTTTG CTAATTCTTTG CCTTG CCTGTATG ATTTATTG G ATGTTG G AATCG CCTG ATG CG GTATTTTCTCCTTACG C ATCTGTG CG GTAT TTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACTATGGT GCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGC TTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCG AAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGG CACTTTTCG G G G AAATGTG CGCG G AACCCCTATTTGTTTATTTTTCTAAATACATTCAA ATATGTATCCG CTCATG AG AC AATAACCC TGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGCCATATTCAACGGGAAACGTCGAGGCCGCGATTAAATTCCAACAT GGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGCTTGTATGGGAAGCCCG ATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGAC GGAATTTATGCCACTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGAAAAAC AGCGTTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCACTCGA TTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGCCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATG CGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACCGGA TTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGT CGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTC AAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAACTGTCAGACCAAGTTTA CTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAA TCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGC GTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGA AGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTA GCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTC AAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACC TACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGT AAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTC GCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTT TACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTT TGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATA CGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCC AACAGTTGCGCAGCCTGAATGGCGAATGGCGATTCCGTTGCAATGGCTGGCGGTAATATTGTTCTGGATATTACCAGCAAGGCCG ATAGTTTG AGTTCTTCTACTCAG G CA AGTG ATGTTATTACTAATCA AAG AAGTATTG CG ACAACG GTTAATTTGCGTG ATG G ACAG ACTCTTTTACTCGGTGGCCTCACTGATTATAAAAACACTTCTCAGGATTCTGGCGTACCGTTCCTGTCTAAAATCCCTTTAATCGGCC TCCTGTTTAGCTCCCGCTCTGATTCTAACGAGGAAAGCACGTTATACGTGCTCGTCAAAGCAACCATAGTACGCGCCCTGTAGCGG CG CATTAAG CG CG G CG G GTGTG GTG GTTACG CG CAG CGTG ACCG CTACACTTG CCAGCG CCCTAGCG CCCG CTCCTTTCG CTTTC TTCCCTTCCTTTCTCG CCACGTTCG CCG G CTTTCCCCGTCAAG CTCTAAATCG G G G G CTCCCTTTAG G GTTCCG ATTTAGTG CTTTA CGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGA CGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATA AG G G ATTTTGCCG ATTTCG G CCTATTG GTTAAAAAATG AG CTG ATTTAACA AAAATTTA ACG CG AATTTTAACAAA ATATTAACG CT TACAATTTAAATATTTGCTTATACAATCTTCCTGTTTTTGGGGCTTTTCTGATTATCAACCGGGGTACATATGATTGACATGCTAGTTT TACGATTACCGTTCATCGCC8. AAV.U7.17ESE3, antisense plasmid sequence (5918 bp) SEQ ID NO: 79GGCGATGAACGGTAATCGTAAAACTAGCATGTCAATCATATGTACCCCGGTTGATAATCAGAAAAGCCCCAAAAACAGGAAGATT GTATAAGCAAATATTTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATA GGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAA TCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAA GCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGC TGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTACTATGGTTGCTTTGACGAGCACGTATAACGTGC TTTCCTCGTTAGAATCAGAGCGGGAGCTAAACAGGAGGCCGATTAAAGGGATTTTAGACAGGAACGGTACGCCAGAATCCTGAG AAGTGTTTTTATAATCAGTGAGGCCACCGAGTAAAAGAGTCTGTCCATCACGCAAATTAACCGTTGTCGCAATACTTCTTTGATTAG TAATAACATCACTTGCCTGAGTAGAAGAACTCAAACTATCGGCCTTGCTGGTAATATCCAGAACAATATTACCGCCAGCCATTGCAA CGGAATCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGCATT AATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGG TCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGA ACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCT GACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGA AGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTC TCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCC GACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTA ACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAG TATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGG TAG CG GTG GTTTTTTTGTTTG CAAG CAG C AG ATTACG CG CAG AAAAAA AG G ATCTCA AG A AG ATCCTTTG ATCTTTTCTACG G G G TCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAA TTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTAGAAAAACTCATCGAGCATCAAATGAAA CTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTT CCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAAT AAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCAGACTTGT TCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGGCG AAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAGTGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAAT ATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAACGCTGTTTTTCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGG AGTACG G ATA AAATG CTTG ATG GTCG G AAGTG G CATAAATTCCGTCAG CC AGTTTAGTCTG ACCATCTCATCTGTAACATC ATTG G C AACG CTACCTTTG CCATGTTTCAG AAACAACTCTG GCG CATCG G G CTTCCC ATACA AG CG ATAG ATTGTCG CACCTG ATTG CCCG A CATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTCGACGTTTCCCGTTGAATATGGCT CATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATA AACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAA AATAGGCGTATCACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACG GTCACAG CTTGTCTGTAAG CG G ATG CCG GG AG CAG ACAAG CCCGTCAG G G CG CGTC AGCG G GTGTTG G CG G GTGTCG G G G CT GGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATAGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGCGG CATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGC G ATTCCAAC ATCCA ATAA ATCATACAGG CAAG G CAAAG AATTAG CAAAATTAAG CAATAA AG CCTCAG AG CATAA AG CTAAATCG GTTGTACCAAAAACATTATGACCCTGTAATACTTTTGCGGGAGAAGCCTTTATTTCAACGCAAGGATAAAAATTTTTAGAACCCTCA TATATTTTAAATGCAATGCCTGAGTAATGTGTAGGTAAAGATTCAAACGGGTGAGAAAGGCCGGAGACAGTCAAATCACCATCAAT ATGATATTCAACCGTTCTAGCTGATAAATTCATGCCGGAGAGGGTAGCTATTTTTGAGAGGTCTCTACAAAGGCTATCAGGTCATTG CCTGAGAGTCTGGAGCAAACAAGAGAATCGCCGGGGGGGGGGGGGGGGGGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCAC TGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGG AGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTAGCCATGCTCTAGTCTAGCC ACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTC ATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTACTGTG TTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACAC CCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTC TTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTA CATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATCAGCGA GCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCGCAGCTTTT AGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACCGCGGTGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCT GGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTA CACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATGGGGC GATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGGGAC TCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTG GGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCTAGTA GAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGC CATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGCGAAG CAG CGCAAA ACG CCTAACCCTAAG CAG ATTCTTC ATG CAATTGTCG GTCA AG CCTTG CCTTGTTGTAG CTTAAATTTTG CTCG CG C ACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAGGCG GAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTGG CCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGATCCT TGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCGCT TGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACCACGCGTGAATTGAATT CAGATCCACGCGTGAATTCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGG GCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAG9. AAV.U7.17SDS, sense plasmid sequence (5923 bp) SEQ ID NO: 80CTGCGCG CTCG CTCG CTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGAATTCACGCGTGGATCTGAATTCAATTCACGCGTGGTACCCTTAAGTCTAGAGGCTCGAG AAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTCGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGCTCTGATCAAGAGA CAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGTTGACATTGATTATTGACTAGTTATTAA TAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACGTCGACGGATCGGGAGATCTCCCGA TCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGAC CGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTT AAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTT CTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCC GCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAAT CTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCG CTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCC ACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTA ACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCTAGCTATCGCCATGTAAG CCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACATTCATCCGGGGTCAGC ACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAGAGTCGACGAGGAA CTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCAC CGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGG ACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAGAAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGT TTTCAG CCA ATCAG CACTG ACTCATTTG CATAG CCTTTACA AG CG GTC ACA AACTC AAG AAACG AG CG GTTTTAATAGTCTTTTAG AATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCAC CCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATCTCACCTTTTCCTAATTTCAGAATAATT TTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGC TCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGCTAGACTAGAGCATGGCTACGTAGA TAAGTAGCATGGCGGGTTAATCATTAACTACAAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCG CTCG CTCG CTCA CTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGG GAGTGGCCCCCCCCCCCCCCCCCCCGGCGATTCTCTTGTTTGCTCCAGACTCTCAGGCAATGACCTGATAGCCTTTGTAGAGACCT CTCAAAAATAGCTACCCTCTCCGGCATGAATTTATCAGCTAGAACGGTTGAATATCATATTGATGGTGATTTGACTGTCTCCGGCCT TTCTCACCCGTTTGAATCTTTACCTACACATTACTCAGGCATTGCATTTAAAATATATGAGGGTTCTAAAAATTTTTATCCTTGCGTTG AAATA AAG G CTTCTCCCG C AAAAGTATTAC AG G GTCATAATGTTTTTG GTACAACCG ATTTAG CTTTATG CTCTG AG G CTTTATTG C TTAATTTTGCTAATTCTTTGCCTTGCCTGTATGATTTATTGGATGTTGGAATCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCG GTATTTCACACCGCATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACTAT GGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACG GGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCA CCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAA CCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGCCATATTCAACGGGAAACGTCGAGGCCGCGATTAAATTCCAA CATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGCTTGTATGGGAAGC CCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCT GACGGAATTTATGCCACTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGAAA AACAGCGTTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCACT CGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGCCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTG ATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACC GGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACG AGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTT TTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAACTGTCAGACCAAGT TTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCA AAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTG CGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTC CGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTC TGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGG ACTCA AG ACG ATAGTTACCG G ATA AGG CGCAG CG GTCG G G CTG AACG G G GG GTTCGTGC AC AC AG CCCAG CTTG G AG CG AAC GACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATC CGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGG GTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGC CTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACC GCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCC CAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCC TTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGCGATTCCGTTGCAATGGCTGGCGGTAATATTGTTCTGGATATTACCAGCAAG GCCGATAGTTTGAGTTCTTCTACTCAGGCAAGTGATGTTATTACTAATCAAAGAAGTATTGCGACAACGGTTAATTTGCGTGATGG ACAGACTCTTTTACTCGGTGGCCTCACTGATTATAAAAACACTTCTCAGGATTCTGGCGTACCGTTCCTGTCTAAAATCCCTTTAATC GGCCTCCTGTTTAGCTCCCGCTCTGATTCTAACGAGGAAAGCACGTTATACGTGCTCGTCAAAGCAACCATAGTACGCGCCCTGTA GCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCG CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTG CTTTACG G CACCTCG ACCCCAAAAAACTTG ATTAG G GTG ATG GTTCACGTAGTG G G CCATCG CCCTG ATAG ACG GTTTTTCG CCC TTTG ACGTTG G AGTCC ACGTTCTTTAATAGTG G ACTCTTGTTCCA AACTG G AAC AACACTC AACCCTATCTCG GTCTATTCTTTTG A TTTATAAG G G ATTTTG CCG ATTTCG G CCTATTG GTTAAAAAATG AGCTG ATTTAACAAA AATTTA ACG CG AATTTTA AC AAAATATT AACGCTTACAATTTAAATATTTGCTTATACAATCTTCCTGTTTTTGGGGCTTTTCTGATTATCAACCGGGGTACATATGATTGACATGC TAGTTTTACGATTACCGTTCATCGCC10. AAV.U7.17SDS, antisense plasmid sequence (5923 bp) SEQ ID NO: 81GGCGATGAACGGTAATCGTAAAACTAGCATGTCAATCATATGTACCCCGGTTGATAATCAGAAAAGCCCCAAAAACAGGAAGATT GTATAAGCAAATATTTAAATTGTAAGCGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATA GGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCC ACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAA TCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAA GCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGC TGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTACTATGGTTGCTTTGACGAGCACGTATAACGTGC TTTCCTCGTTAGAATCAGAGCGGGAGCTAAACAGGAGGCCGATTAAAGGGATTTTAGACAGGAACGGTACGCCAGAATCCTGAG AAGTGTTTTTATAATCAGTGAGGCCACCGAGTAAAAGAGTCTGTCCATCACGCAAATTAACCGTTGTCGCAATACTTCTTTGATTAG TAATAACATCACTTGCCTGAGTAGAAGAACTCAAACTATCGGCCTTGCTGGTAATATCCAGAACAATATTACCGCCAGCCATTGCAA CGGAATCGCCATTCGCCATTCAGGCTGCGCAACTGTTGGGAAGGGCGATCGGTGCGGGCCTCTTCGCTATTACGCCAGCTGCATT AATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGG TCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGA ACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCT GACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGA AGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTC TCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTA ACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAG TATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGG TAG CG GTG GTTTTTTTGTTTG CAAG CAG C AG ATTACG CG CAG AAAAAA AG G ATCTCA AG A AG ATCCTTTG ATCTTTTCTACG G G G TCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAA TTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTAGAAAAACTCATCGAGCATCAAATGAAA CTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTT CCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAAT AAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCAGACTTGT TCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGGCG AAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAGTGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAAT ATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAACGCTGTTTTTCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGG AGTACG G ATA AAATG CTTG ATG GTCG G AAGTG G CATAAATTCCGTCAG CC AGTTTAGTCTG ACCATCTCATCTGTAACATC ATTG G C AACG CTACCTTTG CCATGTTTCAG AAACAACTCTG GCG CATCG G G CTTCCC ATACA AG CG ATAG ATTGTCG CACCTG ATTG CCCG A CATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTCGACGTTTCCCGTTGAATATGGCT CATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATA AACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAA AATAGGCGTATCACGAGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACG GTCACAG CTTGTCTGTAAG CG G ATG CCG GG AG CAG ACAAG CCCGTCAG G G CG CGTC AGCG G GTGTTG G CG G GTGTCG G G G CT GGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATAGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGCGG CATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGC G ATTCCAAC ATCCA ATAA ATCATACAGG CAAG G CAAAG AATTAG CAAAATTAAG CAATAA AG CCTCAG AG CATAA AG CTAAATCG GTTGTACCAAAAACATTATGACCCTGTAATACTTTTGCGGGAGAAGCCTTTATTTCAACGCAAGGATAAAAATTTTTAGAACCCTCA TATATTTTAAATGCAATGCCTGAGTAATGTGTAGGTAAAGATTCAAACGGGTGAGAAAGGCCGGAGACAGTCAAATCACCATCAAT ATGATATTCAACCGTTCTAGCTGATAAATTCATGCCGGAGAGGGTAGCTATTTTTGAGAGGTCTCTACAAAGGCTATCAGGTCATTG CCTGAGAGTCTGGAGCAAACAAGAGAATCGCCGGGGGGGGGGGGGGGGGGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCAC TGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGG AGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTAGCCATGCTCTAGTCTAGCC ACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTC ATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTATTCTGAAATTAGGAAAAGG TGAGATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTC CACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCT CGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATG TTGTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATC AGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCGCA GCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACCGCG GTGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGG CTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCG GGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAG G CTTTACACTTTATG CTCCTG CAG G ACTG GTCCACCTAC AACAAAG CTCTC ATCAACCGTG G CTCCCTCACTTTCTG G CTG G ATG AT GGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGC GGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAA GGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACC CTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAAT TTCTG CCATTC ATCCG CTTATTATCACTTATTCAG G CGTAG CACCAG G CGTTTAAG G GCACCA ATA ACTG CCTTAAAAGG CG CG CC GCGAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGC TCGCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGC AGGCGGAACTCCATATATGGGCTATGAACTAATGACCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTAT ATCTGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCA GATCCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGG TTCGCTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTGATATCTTCTCGAGCCTCTAGACTTAAGGGTACCACGCGTGAATTGAATTCAGATCCACGCGTGAATTCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAG2ndgeneration of constructs:1. U7.2xl7SAS, sense sequence (874 bp) SEQ ID NO: 42TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAA GAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAG GGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCA ATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTAC AATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTAT GTGGGATCCCCATGGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAG CGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATT CACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTAC AGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCC AATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTC CTAGGAAACGCGTATGTG2. U7.2xl7SAS, antisense sequence (874 bp) SEQ ID NO: 43CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTT CATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGT CACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTC CACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCT CGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATG TTGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGA AGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTC ACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTG AGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAA AAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACA GCCAATCACAGCTCCTATGTTGTTA3. Stuffer-U7.2xl7SAS, sense sequence (1902 bp) SEQ ID NO: 82GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTG CCAG CTG G G G CG CCCTCTG GTAAG GTTG G G AAG CCCTG C AAAGTAAACTG G ATG G CTTTCTCG CCG CCAAGG ATCTG ATG G CG C AGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGT TGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGAC GTCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTT AAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGC CTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCC GGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGG CGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCAT AGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCA GGGCGCGCCGGGGGGGGGGGCGCTGAGGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTC GACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAG AAGTTWXACGCATGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAA GCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTG ATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCC CTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCCCATGGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGA CTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCG AATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGA GTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTT TCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTG AGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer 2X17SAS4. Stuffer-U7.2xl7SAS, antisense sequence (1902 bp) SEQ ID NO: 83CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGC TGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAG CTCCTATGTTGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGG AGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCT CCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTT CGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATA AACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCT TGCTGTCCTGCCCCACCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCG ACCGGTGGATCGGCCGCCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGGGACTCTGGGGTTCGAAATGACCGACCAAGCGA CGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGG CTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCC AATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCATTCATCCGCTTATTATCACTTATTCAGGC GTAG CACCAG G CGTTTAAG G G CACCA ATA ACTG CCTTAAAAG GCGCGCCGCGAAGCAGCG CAAA ACG CCTAACCCTAAG CAG A TTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTCGCGCACTACTCAGCGACCTCCAACACACAAGC AGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAGGCGGAACTCCATATATGGGCTATGAACTAATGA CCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGATCCTTGGCGGCGAGAAAGCCATCCAGTTTACT TTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCGCTTGCTGTCCATAAAACCGCCCAGTAGAAG CTG C AGTTG ATATCTTCTCG AG CCTCTAG ACTTA AG G GTACCStuffer 2X17SAS5. U7.17SAS-Stuffer-U7.17SAS, sense sequence (1936 bp) SEQ ID NO: 84TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTC AAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTG GAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTC CGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTG GTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCGGCTCGAGAAGATATCGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTC GCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCA GAAATTCGCCGGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTAC TAGGGTGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCC AACCTTTCATAGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGA GTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAA GTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTG TCGTGCCCGCCCAGTCTAGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGA TAGCCCAGTAGCTGACATTCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCC ACCGCGGTGGCGGCCTAGAGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTC GCTGATCAGTGGGGGGTGGGGTGGGGCAGGACCAATAGCAGCTGCAGAAGTTTAAACGCATGCCATGGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGT TTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAA TGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAG CCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAG CAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer 1X17SAS6. U7.17SAS-Stuffer-U7.17SAS, antisense sequence (1936 bp) SEQ ID NO: 85CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGC TGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAGCTCCTATGTTGTTACCATGGCATGCGTTTAAACTTCTGCAGCTGCTATTGGTCCTGCCCCACCCCACCCCCCACTGATCAGCGAGCT CTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCGCAGCTTTTAGA GCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACCGCGGTGGCTGA TCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGG ACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACA CTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATGGGGCGA TTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGGGACTC TGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGG GCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCA TTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGCGAAGCA GCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCGATATCTTCTCGAGCCGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTG AAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAA GTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCA CTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAG TTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAStuffer 1X17SAS7. aMHCEnh-U7.17SAS-Stuffer-aMHCEnh-U7.17SAS, sense sequence (1914 bp) SEQ ID NO: 86CCTTCAGATTAAAAATAACTGAGGTAAGGGCCTGGGTAGGGGAGGTGGTGTGAGACGCTCCTGTCTCTCCTCTATCTGCCCATCG GCCCTTTGGGGAGGAGGAATGTGCCCAAGGACTAAAAAAAGGCCATGGAGCCAGAGGGGCGAGGGCAACAGACCTTTCATGGGCAAACC7’7"GGGGCCC7’GC7"GaccggtTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCAT AGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAAC TGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGG TCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTT TGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAATCT CGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCGCT GAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCCAC GGTTGATGAGAGCTTTGCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTC GTGCCCGCCCAGTCTAGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATA GCCCAGTAGCTGACATTCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCAAC TGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCCGCCACCAGCGGCCATCAAGCAC GTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGAC AATAGCAGCCTTCAGATTAAAAATAACTGAGGTAAGGGCCTGGGTAGGGGAGGTGGTGTGAGACGCTCCTGTCTCTCCTCTATCT GCCCATCGGCCCTTTGGGGAGGAGGAATGTGCCCAAGGACTAAAAAAAGGCCATGGAGCCAGAGGGGCGAGGGCAACAGACC TTTCATGGGCAAACCTTGGGGCCCTGCTGctgcagaagtttaaacgcatgcTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAAT CAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTT ATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCG AAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGA GCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCC GGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG aMHC enhancer Stuffer1X17SAS8. aMHCEnh-U7.17SAS-Stuffer-aMHCEnh-U7.17SAS, antisense sequence (1914 bp) SEQ ID NO: 87 CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGC TGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAG CTCCTATGTTGTTAgcatgcgtttaaacttctgcagCAGCAGGGCCCCAAGG7’7TGCCCATGAAAGG7’C7GT7GCCC7’CGCCCCTCTGGC7’ CCATGGCCTTTTTTTAGTCCTTGGGCACATTCCTCCTCCCCAAAGGGCCGATGGGCAGATAGAGGAGAGACAGGAGCGTCTCACA CCACCTCCCCTACCCAGGCCCTTACCTCAG7TATTTTTAATCTGAAGGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCC CACCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGGGATCGGCCGCG GGTACAATTCCGCAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTGCTGATCCCGCCTCGAACCAGACACGTAGAAA GCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAG AAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCA GTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGCAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGA TGGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAG CGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAA AGGTTGGGCTTCGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGG GAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAA ATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGA GGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAAT ATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCT GAAAACAGCCAATCACAGCTCCTATGTTGTTAaccggtCAGCAGGGCCCCAAGGTTTGCCCATGAAAGGT’CreTTGCCCT’CGCCCC TCTGGCTCCATGGCCTTTTTTTAGTCCTTGGGCACATTCCTCCTCCCCAAAGGGCCGATGGGCAGATAGAGGAGAGACAGGAGCG TCTCACACCACCTCCCCTACCCAGGCCCTTACCTCAGTTATTTTTAATCTGAAGG aMHC enhancerStuffer1X17SAS9. U7.4xl7SAS with stuffer, sense sequence (1944 bp) SEQ ID NO: 88GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACC GTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTG CAGAAGTTTAAACGCATGTAACAACATAGGAG CTGTG ATTGG CTGTTTTCAGCCA ATCAG CACTG ACTC ATTTG CATAG CCTTTA CAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTT GTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGC TCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAA ACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTT CTCTGGTTTCCTAGGAAACGCGTATGTGGCTAGCGCATGCTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCA CTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGA ACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGT GGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGG TTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGT GTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGAAGCTTAGTACTTAACAACATAGGAGCTGTGATTG GCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGT CTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCT TGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATGGTGACAGCCTGTGAA ATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAG TTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCCCATGGT AACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCA AGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGG AGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCC GCAATGGTGACAGCCTGTGAAATCTGTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGG TCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAAC GCGTATGTGStuffer 4X17SAS10. U7.4xl7SAS with stuffer, antisense sequence (1944 bp) SEQ ID NO: 89CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGC TGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAG CTCCTATGTTGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGG AGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCT CCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTT CGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATA AACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGA TTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAAGTACTAAGCTTCACATACGCGTTTCCTAGGAAACCAGAGAAGGAT CAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTT CCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCC AGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAA GCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAA GG CTATGCA AATG AGTCAGTG CTG ATTGG CTG A AAACAG CCA ATCACAGCTCCTATGTTGTTAG CATG CGCTAG CCACATACG C GTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTA GACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCACAGATTTCACAGGCTGTCACCATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACA CCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGT TTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATCA GCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGGATATCTTCTCGAGCCTC TAGACTTAAGGGTACCStuffer4X17SAS11. U7.2xl7ESEl, sense sequence (870 bp) SEQ ID NO: 46TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAA GAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAG GGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCA ATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAA TGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGT GGGATCCCCATGGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCG GTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCA CATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAG ACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTT CACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAG GAAACGCGTATGTG12. U7.2xl7ESEl, antisense sequence (870 bp) SEQ ID NO: 47CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTT CATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGA CAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCA CACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCG TTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTT GTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAA GAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCA GCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGA TCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAG ACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCA ATCACAGCTCCTATGTTGTTA13. Stuffer-U7.2xl7ESEl, sense sequence (1898 bp) SEQ ID NO: 90GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTG CCAG CTG G G G CG CCCTCTG GTAAG GTTG G G AAG CCCTG C AAAGTAAACTG G ATG G CTTTCTCG CCG CCAAGG ATCTG ATG G CG C AGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGT TGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGAC GTCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTT AAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGC CTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCC GGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGG CGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCAT AGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCA GGGCGCGCCGGGGGGGGGGGCGCTGAGGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTC GACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAG AAGTTTAAACGCATGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTG ATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCG CTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCT CCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTG GTTTCCTAGGAAACGCGTATGTGGGATCCCCATGGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGAC TCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGA ATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGT TGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGA CTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAG GGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer 2X17ESE114. Stuffer-U7.2xl7ESEl, antisense sequence (1898 bp) SEQ ID NO: 91CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACAC AGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCAT TTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAAC CGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCT CCTATGTTGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGA GCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTC CAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGA TGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAAC AATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGC TGTCCTGCCCCACCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACC GGTGGATCGGCCGCCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGC CCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTG GATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATT ATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAG CACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGCGAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTT CATG CAATTGTCG GTCA AG CCTTG CCTTGTTGTAG CTTAAATTTTG CTCG CGC ACTACTCAG CG ACCTCCA AC AC ACA AG CAG G G AGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAGGCGGAACTCCATATATGGGCTATGAACTAATGACCCC GTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGATCCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGC AGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCGCTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGC AGTTG ATATCTTCTCG AG CCTCTAG ACTTAAG G GTACCStuffer 2X17ESE115. U7.17ESE1-Stuffer-U7.17ESE1, sense sequence (1932 bp) SEQ ID NO: 92TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTC AAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTG GAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTC CGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTC TACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCGGCTCGAGAAGATATCGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCG GCGCGCCTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAG GGTGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAAC CTTTCATAGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTC CCGCTCAGGGCGCGCCGGGGGGGGGGGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCG CCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGT GTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTC GTGCCCGCCCAGTCTAGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATA GCCCAGTAGCTGACATTCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCAC CGCGGTGGCGGCCTAGAGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAA GCTGCGGAATTGTACCCGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGC TGATCAGTGGGGGGTGGGGTGGGGCAGGACCAATAGCAGCTGCAGAAGTTTAAACGCATGCCATGGTAACAACATAGGAGCT GTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTT TAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATG GCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAA CAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer 1X17ESE116. U7.17ESE1-Stuffer-U7.17ESE1, antisense sequence (1932 bp) SEQ ID NO: 93CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACAC AGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCAT TTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAAC CGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTACCATGGCATGCGTTTAAACTTCTGCAGCTGCTATTGGTCCTGCCCCACCCCACCCCCCACTGATCAGCGAGCTCT AGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCGCAGCTTTTAGAG CAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACCGCGGTGGCTGATC CCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGAC AAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGGCACGACA GGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTT TATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATGGGGCGATTC AGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGGGACTCTGG GGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGGCTT CGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCTAGTAGAGCT TTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCATTCA TCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGCGAAGCAGCG CAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCGATATCTTCTCGAGCCGGATCCCACATACGCGTTTCCTAGGA AACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAA TTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATA TGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGT GACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAStuffer1X17ESE117. aMHCEnh-U7.17ESEl-Stuffer-aMHCEnh-U7.17ESEl, sense sequence (1910 bp) SEQ ID NO: 94CCTTCAGATTAAAAATAACTGAGGTAAGGGCCTGGGTAGGGGAGGTGGTGTGAGACGCTCCTGTCTCTCCTCTATCTGCCCATCG GCCCTTTGGGGAGGAGGAATGTGCCCAAGGACTAAAAAAAGGCCATGGAGCCAGAGGGGCGAGGGCAACAGACCTTTCATGG GCAAACCTTGGGGCCCTGCrGaccggtTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCAT AGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAAC TGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCT TCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCG GAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGA TCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAATCTCGT GATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCGCTGAG GTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGGT TGATGAGAGCTTTGCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTG CCCGCCCAGTCTAGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCC CAGTAG CTG AC ATTCATCCG G G GTCAG CACCGTTTCTG CG G ACTG G CTTTCTACGTGTCTG GTTCG AG G CG G G ATCAG CAACTG GCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCCGCCACCAGCGGCCATCAAGCACGT TATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAA TAGCAGCCTTCAGATTAAAAATAACTGAGGTAAGGGCCTGGGTAGGGGAGGTGGTGTGAGACGCTCCTGTCTCTCCTCTATCTGC CCATCGGCCCTTTGGGGAGGAGGAATGTGCCCAAGGACTAAAAAAAGGCCATGGAGCCAGAGGGGCGAGGGCAACAGACCTTT CATGGGCAAACCT'TGGGGCCCTGCT’GctgcagaagtttaaacgcatgcTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCA GCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTAT CGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAA AGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTG TGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG aMHC enhancer Stuffer 1X17ESE118. aMHCEnh-U7.17ESEl-Stuffer-aMHCEnh-U7.17ESEl, antisense sequence (1910 bp) SEQ ID NO: 95 CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACAC AGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCAT TTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAAC CGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCT CCTATGTTGTTAgcatgcgtttaaacttctgcagCAGC GGGCCCC AGGT7TGCCCA7G AAGGTC7G7~7GCCCTCGCCCC7'C7'GGC7'CC ATGGCCTTTTTTTAGTCCTTGGGCACATTCCTCCTCCCCAAAGGGCCGATGGGCAGATAGAGGAGAGACAGGAGCGTCTCACACC ACCTCCCCTACCCAGGCCCT7ACCTCAG7TATTTTTAATC7~GAAGGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCA CCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGGGATCGGCCGCGGG TACAATTCCGCAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTGCTGATCCCGCCTCGAACCAGACACGTAGAAAGC CAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAA AGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTG AGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGCAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATGG GGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGG GACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGG TTGGGCTTCGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAA GAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTC AGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTG AGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTA AAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAA- Ill -CAGCCAATCACAGCTCCTATGTTGTTAaccggtCAGCAGGGCCCCAAGGTTTGCCCATGAAAGGTCTGTTGCCCTCGCCCCTCTGGCTCCATGGCCTTTTTTTAGTCCTTGGGCACATTCCTCCTCCCCAAAGGGCCGATGGGCAGATAGAGGAGAGACAGGAGCGTCTCACACCACCTCCCCTACCCAGGCCCTTACCTCAGTTATTTTTAATCTGAAGG aMHC enhancer Stuffer 1X17ESE119. U7.4xl7ESEl with stuffer, sense sequence (1936 bp) SEQ ID NO: 96GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACC GTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTG CAGAAGTTTAAACGCATGTAACAACATAGGAG CTGTG ATTGG CTGTTTTCAGCCA ATCAG CACTG ACTC ATTTG CATAG CCTTTA CAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTT GTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGC TCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACC CCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTC TGGTTTCCTAGGAAACGCGTATGTGGCTAGCGCATGCTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTG ACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACC GAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGG AGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTC TGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGA GAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGAAGCTTAGTACTTAACAACATAGGAGCTGTGATTGGCTG TTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTT AGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATC TCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGTCTGTGTTAGTGATGGCTGA ATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTC CCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCCCATGGTAACAACA TAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACG AGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTG TGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAATTGT CTGTGTTAGTGATGGCTGAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGA AAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer 4X17ESE120. U7.4xl7ESEl with stuffer, antisense sequence (1936 bp) SEQ ID NO: 97CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACAC AGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCAT TTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAAC CGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCT CCTATGTTGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGA GCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTC CAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGA TGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAAC AATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTG GCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAAGTACTAAGCTTCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAA AGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCG ACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCAC AGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTA TGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAGCATGCGCTAGCCACATACGCGTTTCC TAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAG TGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTCAGCCATCACTAACACAGACAATTGCGGAA GTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCA CTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAG TTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTACAT GCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATCAGCGAGCT CTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGGATATCTTCTCGAGCCTCTAGACTT AAGGGTACCStuffer 4X17ESE121. U7.2X17ESE2, sense sequence (872 bp) SEQ ID NO: 50TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAA GAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAG GGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCA ACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACA ATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATG TGG G ATCCCC ATG GTAACAACATAG G AG CTGTG ATTG G CTGTTTTC AG CCA ATCAG CACTG ACTC ATTTG CATAG CCTTTACAAG C GGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTC ACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACA GACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAA TTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCT AGGAAACGCGTATGTG22. U7.2X17ESE2, antisense sequence (872 bp) SEQ ID NO: 51CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTT CATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATC CTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCC ACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTC GTTTCTTG AGTTTGTG ACCG CTTGTAA AG G CTATG CAAATG AGTCAGTG CTG ATTG G CTG AAAACAG CC AATCACAG CTCCTATGT TGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAA GAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGA CCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGA GATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAA AGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAG CCAATC AC AG CTCCTATGTTGTTA23. Stuffer-U7.2xl7ESE2, sense sequence (1900 bp) SEQ ID NO: 98GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTG CCAG CTG G G G CG CCCTCTG GTAAG GTTG G G AAG CCCTG C AAAGTAAACTG G ATG G CTTTCTCG CCG CCAAGG ATCTG ATG G CG C AGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGT TGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGAC GTCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTT AAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGC CTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCC GGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCAT AGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCA GGGCGCGCCGGGGGGGGGGGCGCTGAGGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTC GACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAG AAGTTWXACGCATGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAA GCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTG ATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCG CTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCC TCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTG GTTTCCTAGGAAACGCGTATGTGGGATCCCCATGGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGAC TCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGA ATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGT TGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGA GGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer2X17ESE224. Stuffer-U7.2xl7ESE2, antisense sequence (1900 bp) SEQ ID NO: 99CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACA GATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAG CTCCTATGTTGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGG AGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCT CCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTT CGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATA AACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCT TGCTGTCCTGCCCCACCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCG ACCGGTGGATCGGCCGCCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGGGACTCTGGGGTTCGAAATGACCGACCAAGCGA CGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGG CTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCC AATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCATTCATCCGCTTATTATCACTTATTCAGGC GTAG CACCAG G CGTTTAAG G G CACCA ATA ACTG CCTTAAAAG GCGCGCCGCGAAGCAGCG CAAA ACG CCTAACCCTAAG CAG A TTCTTCATGCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTCGCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGCAGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAGGCGGAACTCCATATATGGGCTATGAACTAATGA CCCCGTAATTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTGGCCCGTACATCGCGAAGAGTCAAGAACGC GAAACGATCCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGATCCTTGGCGGCGAGAAAGCCATCCAGTTTACT TTGCAGGGCTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCGCTTGCTGTCCATAAAACCGCCCAGTAGAAG CTG C AGTTG ATATCTTCTCG AG CCTCTAG ACTTA AG G GTACCStuffer2X17ESE225. U7.17ESE2-Stuffer-U7.17ESE2, sense sequence (1934 bp) SEQ ID NO: 100TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTC AAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCGGCTCGAGAAGATATCGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGCCTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCCGGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCGCTGAGGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAAAGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCCGCCCAGTCTAGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCCAGTAGCTGACATTCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGCGGTGGCGGCCTAGAGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACCAATAGCAGCTGCAGAAGTTTAAACGCATGCCATGGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTG TTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCA AAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer1X17ESE226. U7.17ESE2-Stuffer-U7.17ESE2, antisense sequence (1934 bp) SEQ ID NO: 101CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAGCTCCTATGTTGTTACCATGGCATGCGTTTAAACTTCTGCAGCTGCTATTGGTCCTGCCCCACCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGCGGGTACAATTCCGCAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTAACTTTCTGGTTTTTCAGTTCCTCGTCGACTCTAGGCCGCCACCGCGGTGGCTGATCCCGCCTCGAACCAGACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTCCTGCAGGACTGGTCCACCTACAACAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGATGGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGATGATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATGTCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCACCAGGCGTTTAAGGGCACCAATAACTGCCTTAAAAGGCGCGCCGCGAAGCAGCGCAAAACGCCTAACCCTAAGCAGATTCTTCATGCAATTGTCGGTCGATATCTTCTCGAGCCGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCA CTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAG TTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAStuffer1X17ESE227. aMHCEnh-U7.17ESE2-Stuffer-aMHCEnh-U7.17ESE2, sense sequence (1912 bp) SEQ ID NO: 102CCTTCAGATTAAAAATAACTGAGGTAAGGGCCTGGGTAGGGGAGGTGGTGTGAGACGCTCCTGTCTCTCCTCTATCTGCCCATCG GCCCTTTGGGGAGGAGGAATGTGCCCAAGGACTAAAAAAAGGCCATGGAGCCAGAGGGGCGAGGGCAACAGACCTTTCATGG GC AACCTTGGGGCCCTGCreaccggtTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCAT AGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAAC TGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCT TCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTC GGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTG ATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAATCTCG TGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCAGGGCGCGCCGGGGGGGGGGGCGCTGA GGTCTGCCTCGTGAAGAAGGTGTTGCTGACTCATACCAGGCCTGAATCGCCCCATCATCCAGCCAGAAAGTGAGGGAGCCACGG TTGATGAGAGCTTTGCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGT GCCCGCCCAGTCTAGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGC CCAGTAGCTGACATTCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCAACTG GCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCTGCGGAATTGTACCCGCGGCCGATCCCGCCACCAGCGGCCATCAAGCACGT TATCGATACCGTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAA TAGCAGCCTTCAGATTAAAAATAACTGAGGTAAGGGCCTGGGTAGGGGAGGTGGTGTGAGACGCTCCTGTCTCTCCTCTATCTGC CCATCGGCCCTTTGGGGAGGAGGAATGTGCCCAAGGACTAAAAAAAGGCCATGGAGCCAGAGGGGCGAGGGCAACAGACCTTT CA7GGGCAAACC7TGGGGCCC7GC7GctgcagaagtttaaacgcatgcTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCA GCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTAT CGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAA AGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCA GGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGT GTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTG aMHC enhancer Stuffer 1X17ESE228. aMHCEnh-U7.17ESE2-Stuffer-aMHCEnh-U7.17ESE2, antisense sequence (1912 bp) SEQ ID NO: 103 CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACA GATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAG CTCCTKVGTVGlJAgcatgcgmaaactctgcagCAGCAGGGCCCCAAGGTTTGCCCATGAAAGGTCTGTTGCCCTCGCCCCTCTGGCT CCATGGCCTTTTTTTAGTCCTTGGGCACATTCCTCCTCCCCAAAGGGCCGATGGGCAGATAGAGGAGAGACAGGAGCGTCTCACA CCACCTCCCCTACCCAGGCCCTTACC7'CAG7~TATTT7~TAATCTGAAGGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCC CACCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGGGATCGGCCGCG GGTACAATTCCGCAGCTTTTAGAGCAGAAGTAACACTTCCGTACAGGCCAGTTGCTGATCCCGCCTCGAACCAGACACGTAGAAA GCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAG AAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGGCACGACAGGTTTCCCGACTGGAAAGCGGGCA GTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGCAAAGCTCTCATCAACCGTGGCTCCCTCACTTTCTGGCTGGATGA TGGGGCGATTCAGGCCTGGTATGAGTCAGCAACACCTTCTTCACGAGGCAGACCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAG CGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCAACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAA AGGTTGGGCTTCGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGG GAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGA GGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAAT ATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCT GAAAACAGCCAATCACAGCTCCTATGTTGTTAaccggtCAGCAGGGCCCCA4GG7TTGCCG47GA4AGGTC7G7TGCCCTCGCCCC TCTGGCTCCATGGCCTTTTTTTAGTCCTTGGGCACATTCCTCCTCCCCAAAGGGCCGATGGGCAGATAGAGGAGAGACAGGAGCG TCTCACACCACCTCCCCTACCCAGGCCCTTACCTCAGTTATTTTTAATCTGAAGG aMHC enhancer Staffer 1X17ESE229. U7.4xl7ESE2 with staffer, sense sequence (1940 bp) SEQ ID NO: 104GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACC GTCGACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTG CAGAAGTTTAAACGCATGTAACAACATAGGAG CTGTG ATTGG CTGTTTTCAGCCA ATCAG CACTG ACTC ATTTG CATAG CCTTTA CAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTT GTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGC TCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAAC CCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCT CTGGTTTCCTAGGAAACGCGTATGTGGCTAGCGCATGCTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACT GACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAAC CGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTG GAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTT TCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTG AGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGAAGCTTAGTACTTAACAACATAGGAGCTGTGATTGGCT GTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTT TTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGA TCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACCAGGATCTGTTCCCTTGTGG TCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTC TTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCCCATGGTAACA ACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAAGAA ACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAGGG GTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAA CCAGGATCTGTTCCCTTGTGGTCAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACA ATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTAT GTGStaffer 4X17ESE230. U7.4X17ESE2 with staffer, antisense (1940 bp) SEQ ID NO: 105CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACA GATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCC ATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAA ACCG CTCGTTTCTTG AGTTTGTG ACCG CTTGTA AAGG CTATGCA AATG AGTCAGTG CTG ATTGG CTGAA AAC AG CCA ATCACAG CTCCTATGTTGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGG AGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCT CCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTT CGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGA TTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTAAGTACTAAGCTTCACATACGCGTTTCCTAGGAAACCAGAGAAGGAT CAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTT CCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGTTGCGGAAGTGCGTCTGTAGCGAGCC AGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAA GCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAA GG CTATGCA AATG AGTC AGTG CTG ATTGG CTG A AAACAG CCA ATCACAGCTCCTATGTTGTTAG CATG CGCTAG CCACATACG C GTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTGACCACAAGGGAACAGATCCTGGT TGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACA CCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGT TTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCACTGATCA GCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGTGGATCGGCCGGATATCTTCTCGAGCCTC TAGACTTAAGGGTACCStuffer 4X17ESE231. U7.2X17ESE3, sense sequence (866 bp) SEQ ID NO: 54TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTCAA GAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTGGAG GGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTACAATGA AAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGG ATCCCC ATG GTAACAACATAG G AG CTGTG ATTG G CTGTTTTCAG CCA ATCAG CACTG ACTCATTTG CATAG CCTTTAC AAG CG GTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATA TCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGC ACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGG TCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACG CGTATGTG32. U7.2X17ESE3, antisense sequence (866 bp) SEQ ID NO: 55CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCTTT CATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTACTGT GTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTTCCACA CCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCGCTCGTTT CTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCCTATGTTGTT ACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAG AACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAG AGGCAGATTACTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAA G GTG CC ATTTCCACACCCCTCCACTG ATATGTG AATC ACA AAG CACAGTTCCTTATTCG GTTCG ATA AACA ATATTCTAAAAG ACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATC ACAG CTCCTATGTTGTTA33. Stuffer-U7.2xl7ESE3, sense sequence (1894 bp) SEQ ID NO: 106GGTACCCTTAAGTCTAGAGGCTCGAGAAGATATCAACTGCAGCTTCTACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTG CCAG CTG G G G CG CCCTCTG GTAAG GTTG G G AAG CCCTG C AAAGTAAACTG G ATG G CTTTCTCG CCG CCAAGG ATCTG ATG G CG C AGGGGATCAAGCTCTGATCAAGAGACAGGATGAGGATCGTTTCGCGTTCTTGACTCTTCGCGATGTACGGGCCAGATATACGCGT TGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTCCGCCTGCAGGGACGTCGACGGATCGGGAGATCTCCCGATCCCCTATCTGCTCCCTGCTTGTGTGTTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTT AAGCTACAACAAGGCAAGGCTTGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGCGCGC CTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAATTCGCC GGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGGTGGG CGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCAT AGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTCA GGGCGCGCCGGGGGGGGGGGCGCTGAGGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTC GACTAGAGCTCGCTGATCAGTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGCTGCAG AAGTTWXACGCATGTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAA GCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTG ATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCG CTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCC AATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTT CCTAGGAAACGCGTATGTGggatccccatggTAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTG CATAGCCTTTACAAGCGGTCACAAACTCAAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGG AACTGTGCTTTGTGATTCACATATCAGTGGAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGT CCTTCCCTGGCTCGCTACAGACGCACTTCCGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTC GGAAAACCCCTCCCAATTTCACTGGTCTACAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTG ATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGStuffer2X17ESE334. Stuffer-U7.2xl7ESE3, antisense sequence (1894 bp) SEQ ID NO: 107CACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGCGGGGAAGAGAACTGTTTTGCT TTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCAAAAATTAGAAGAGGCAGATTA CTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGAGGGTGAGATCAAGGTGCCATTT CCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAATATTCTAAAAGACTATTAAAACCG CTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCTGAAAACAGCCAATCACAGCTCC TATGTTGTTACCATGGGGATCCCACATACGCGTTTCCTAGGAAACCAGAGAAGGATCAAAGCCCCTCTCACACACCGGGGAGC GGGGAAGAGAACTGTTTTGCTTTCATTGTAGACCAGTGAAATTGGGAGGGGTTTTCCGACCGAAGTCAGAAAACCTGCTCCA AAAATTAGAAGAGGCAGATTACTGTGTTGCGGAAGTGCGTCTGTAGCGAGCCAGGGAAGGACATCAACTCCACTTTCGATGA GGGTGAGATCAAGGTGCCATTTCCACACCCCTCCACTGATATGTGAATCACAAAGCACAGTTCCTTATTCGGTTCGATAAACAAT ATTCTAAAAGACTATTAAAACCGCTCGTTTCTTGAGTTTGTGACCGCTTGTAAAGGCTATGCAAATGAGTCAGTGCTGATTGGCT G AA AAC AG CCA ATCACAG CTCCTATGTTGTTACATGCGTTTAAACTTCTGCAGCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGT CCTGCCCCACCCCACCCCCCACTGATCAGCGAGCTCTAGTCGACGGTATCGATAACGTGCTTGATGGCCGCTGGTGGCGACCGGT GGATCGGCCGCCTCAGCGCCCCCCCCCCCGGCGCGCCCTGAGCGGGACTCTGGGGTTCGAAATGACCGACCAAGCGACGCCCA ACCTGCCATCACGAGATTTCGATTCCACCGCCGCCTTCTATGAAAGGTTGGGCTTCGGAATCGTTTTCCGGGACGCCGGCTGGAT GATCCTCCAGCGCGGGGATCTCATGCTGGAGTTCTTCGCCCACCCTAGTAGAGCTTTAAATCTCTGTAGGTAGTTTGTCCAATTATG TCACACCACAGAAGTAAGGTTCCTTCACAAAGATCCGGCGAATTTCTGCCATTCATCCGCTTATTATCACTTATTCAGGCGTAGCAC CAG G CGTTTAAG G G CACCAATAACTG CCTTAAA AG G CG CG CCG CG AAG CAG CG C AAAACG CCTA ACCCTA AG CAG ATTCTTCAT GCAATTGTCGGTCAAGCCTTGCCTTGTTGTAGCTTAAATTTTGCTCGCGCACTACTCAGCGACCTCCAACACACAAGCAGGGAGC AGATAGGGGATCGGGAGATCTCCCGATCCGTCGACGTCCCTGCAGGCGGAACTCCATATATGGGCTATGAACTAATGACCCCGTAA TTGATTACTATTAATAACTAGTCAATAATCAATGTCAACGCGTATATCTGGCCCGTACATCGCGAAGAGTCAAGAACGCGAAACGAT CCTCATCCTGTCTCTTGATCAGAGCTTGATCCCCTGCGCCATCAGATCCTTGGCGGCGAGAAAGCCATCCAGTTTACTTTGCAGGG CTTCCCAACCTTACCAGAGGGCGCCCCAGCTGGCAATTCCGGTTCGCTTGCTGTCCATAAAACCGCCCAGTAGAAGCTGCAGTTG ATATCTTCTCGAGCCTCTAGACTTAAGGGTACCStuffer2X17ESE335. U7.17ESE3-Stuffer-U7.17ESE3, sense sequence (1928 bp) SEQ ID NO: 108TAACAACATAGGAGCTGTGATTGGCTGTTTTCAGCCAATCAGCACTGACTCATTTGCATAGCCTTTACAAGCGGTCACAAACTC AAGAAACGAGCGGTTTTAATAGTCTTTTAGAATATTGTTTATCGAACCGAATAAGGAACTGTGCTTTGTGATTCACATATCAGTG GAGGGGTGTGGAAATGGCACCTTGATCTCACCCTCATCGAAAGTGGAGTTGATGTCCTTCCCTGGCTCGCTACAGACGCACTTC CGCAACACAGTAATCTGCCTCTTCTAATTTTTGGAGCAGGTTTTCTGACTTCGGTCGGAAAACCCCTCCCAATTTCACTGGTCTA CAATGAAAGCAAAACAGTTCTCTTCCCCGCTCCCCGGTGTGTGAGAGGGGCTTTGATCCTTCTCTGGTTTCCTAGGAAACGCGTATGTGGGATCCGGCTCGAGAAGATATCGACCGACAATTGCATGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGGC GCGCCTTTTAAGGCAGTTATTGGTGCCCTTAAACGCCTGGTGCTACGCCTGAATAAGTGATAATAAGCGGATGAATGGCAGAAAT TCGCCGGATCTTTGTGAAGGAACCTTACTTCTGTGGTGTGACATAATTGGACAAACTACCTACAGAGATTTAAAGCTCTACTAGGG TGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCGGCGTCCCGGAAAACGATTCCGAAGCCCAACCTT TCATAGAAGGCGGCGGTGGAATCGAAATCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCG CTCAGGGCGCGCCGGGGGGGGGGGCGCTGAG GTCTG CCTCGTG A AG A AG GTGTTG CTG ACTCATACCAG G CCTG AATCG CCCC ATCATCCAGCCAGAAAGTGAGGGAGCCACGGTTGATGAGAGCTTTGTTGTAGGTGGACCAGTCCTGCAGGAGCATAAAGTGTAA AGCCTGGGGTGCCTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGC CCGCCCAGTCTAGCTATCGCCATGTAAGCCCACTGCAAGCTACCTGCTTTCTCTTTGCGCTTGCGTTTTCCCTTGTCCAGATAGCCC AGTAGCTGACATTCATCCGGGGTCAGCACCGTTTCTGCGGACTGGCTTTCTACGTGTCTGGTTCGAGGCGGGATCAGCCACCGC GGTGGCGGCCTAGAGTCGACGAGGAACTGAAAAACCAGAAAGTTAACTGGCCTGTACGGAAGTGTTACTTCTGCTCTAAAAGCT GCGGAATTGTACCCGCGGCCGATCCACCGGTCGCCACCAGCGGCCATCAAGCACGTTATCGATACCGTCGACTAGAGCTCGCTGA TCAGTGGGGGGTGGGGTGGGGCAGGACCAATAGCAGCTGCAGAAGTTTAAACGCATGCCATGGTAACAACATAGGAGCTGTG ATTGGCTGTTTTCAGC...

Claims

CLAIMSWHAT IS CLAIMED IS:1 . A nucleic acid that binds or is complementary to a polynucleotide encoding(a) exon 17 of the DMD gene comprising intronic sequence surrounding exon 17 comprising or consisting of the nucleotide sequence of SEQ ID NO: 1 or 2;(b) exon 17 of the DMD gene or its reverse complementary sequence comprising or consisting of SEQ ID NO: 3 or 4; or(c) the amino acid sequence of SEQ ID NO: 5.

2. The nucleic acid of claim 1 comprising or consisting of:(a) an antisense encoding nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 , or comprising or consisting of the nucleotide sequence of SEQ ID NO: 7, 13, 19, 25 or 31 ;(b) an antisense encoding nucleotide sequence that binds to a target nucleotide sequence of SEQ ID NO: 6, 12, 18, 24, or 30;(c) an antisense encoding nucleotide sequence that binds to a target mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 8, 14, 20, 26, or 32; or(d) an antisense encoding nucleotide sequence that binds to a target antisense mRNA sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 9, 15, 21 , 27, or 33, or comprising or consisting of the nucleotide sequence of 9, 15, 21 , 27, or 33.

3. The nucleic acid of claim 1 or 2 further comprising a promoter.

4. The nucleic acid of any one of claims 1 -3, wherein the promoter is a U6, U7, tRNA, H1 , CMV, minimal CMV, T7, EF1 -alpha, Minimal EF1 -alpha, or a tissue-specific promoter including, but not limited to, a muscle-specific promoter.

5. The nucleic acid of claim 4, wherein the promoter is a U7 promoter.

6. The nucleic acid of claim 5, wherein the nucleotide sequence comprises or consists of(a) a nucleotide sequence encoding a U7snRA comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 10, 16, 22, 28, or 34; or(b) a nucleotide sequence encoding a U7snRNA reverse complement sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of SEQ ID NO: 11 , 17, 23, 29, or 35, or comprising or consisting of the nucleotide sequence of SEQ ID NO: 11 , 17, 23, 29, or 35.

7. The nucleic acid of any one of claims 1 -6 further comprising an inverted terminal repeat region.

8. A nucleic acid comprising a combination of any two or more of the nucleic acids of any one of claims 1-7.

9. The nucleic acid of claim 8 wherein the nucleotide sequence comprises or consists of(a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 42-61 , or(b) a nucleotide sequence comprising any one of SEQ ID NOs: 42-61 .

10. The nucleic acid of claim 1 , wherein the nucleotide sequence comprises or consists of(a) a nucleotide sequence comprising at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleotide sequence of any one of SEQ ID NOs: 62-161 , or(b) a nucleotide sequence comprising any one of SEQ ID NOs: 162-161 .

11. A composition comprising the nucleic acid of any one of claims 1 -10.

12. A vector, nanoparticle, extracellular vesicle, or exosome comprising the nucleic acid of any one of claims 1 -10 or the composition of claim 1 1 .

13. The vector of claim 12, wherein the vector is an adeno-associated virus (AAV).

14. The AAV of claim 13, wherein the AAV lacks rep and cap genes.

15. The AAV of claim 13 or 14, wherein the AAV is a recombinant AAV (rAAV) or a self- complementary recombinant AAV (scAAV).

16. The AAV of any one of claims 13-15, wherein the AAV is AAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV.rh74, AAV.rh8, AAV.rhW, AAV11 , AAV12, AAV13, AAV-anc80, AAV-B1 , AAV-BR1 , AAV.PHP.EB, AAVv66, AAV2 / 1 , AAV2 / 8, AAV2 / 9, AAVMYO, MYOAAV, MYOAAV1 A, MYOAAV2A, MYOAAV3A, or any other myotropic serotype, or any derivative thereof.

17. The AAV of any one of claims 13-16, wherein the AAV is AAVMYO, MYOAAV, MYOAAV1 A, MYOAAV2A, MYOAAV3A, or AAV9.

18. A method for inducing skipping of exon 17 of the DMD gene in a cell, the method comprising providing the cell with(a) the nucleic acid of any one of claims 1-10;(b) the composition of claim 1 1 ;(c) the vector, nanoparticle, extracellular vesicle, or exosome of claim 12; or(d) the AAV of any one of claims 13-17.

19. The method of claim 18, wherein the cell is in a human subject.

20. A method for treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17) comprising administering to the subject an effective amount of(a) the nucleic acid of any one of claims 1-10;(b) the composition of claim 1 1 ;(c) the vector, nanoparticle, extracellular vesicle, or exosome of claim 12; or(d) the AAV of any one of claims 13-17.21 . The method of any one of claims 18-20, wherein the mutation is any mutation involving, surrounding, or affecting DMD exon 17.

22. The method of claim 20 or 21 , wherein administering results in increased expression of dystrophin protein or a functional dystrophin protein in the subject.

23. The method of any one of claims 20-22, wherein administering inhibits the progression of dystrophic pathology in the subject.

24. The method of any one of claims 20-23, wherein administering improves muscle function in the subject.

25. The method of claim 24, wherein the improvement in muscle function is an improvement in muscle strength.

26. The method of claim 24, wherein the improvement in muscle function is an improvement in stability in standing and walking.

27. Use of(a) the nucleic acid of any one of claims 1-10;(b) the composition of claim 1 1 ;(c) the vector, nanoparticle, extracellular vesicle, or exosome of claim 12; or(d) the AAV of any one of claims 13-17. in treating, ameliorating, and / or preventing a muscular dystrophy in a subject with a mutation amenable to skipping exon 17 of the DMD gene (DMD exon 17).

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