Therapeutic nucleic acids for treating genetic disorders

Abstract

There is provided a therapeutic nucleic acid capable of binding to a target indel allele associated with a pathogenic allele, wherein the pathogenic allele causes a dominant negative genetic disorder or gain-of function genetic disorder, and wherein the disorder is not Huntington's disease. There is further provided a conjugate, delivery particle, and pharmaceutical composition thereof and their uses. There is further provided a therapeutic nucleic acid which is capable of binding to a target indel allele of an intronic indel selected from: rs59464879, rs751205475 and rs78373442 associated with a pathogenic HTT allele which causes Huntington's disease, and uses thereof.

Description

THERAPEUTIC NUCLEIC ACIDS FOR TREATING GENETIC DISORDERS FIELD OF THE INVENTION

[0001] The present invention relates to therapeutic nucleic acids that are capable of binding to target indel alleles associated with pathogenic alleles, specifically capable of binding to target indel alleles present in pre-mRNA or DNA where allele-specific targeting could be therapeutic. The present invention further relates to derived products and pharmaceutical compositions thereof, and to the prevention or treatment of such disorders that can benefit from allele-specific targeting using the therapeutic nucleic acids. INTRODUCTION

[0002] Allele-specific targeting of pathogenic alleles in cellular nucleic acids holds significant therapeutic potential. This approach can preserve gene function while simultaneously providing a therapeutic effect by targeting alleles that cause pathology, as well as non- pathogenic alleles whose modulation can have a therapeutic effect or enhance the efficacy of treatments.

[0003] Dominant genetic disorders are those which result from a mutation in a gene which is dominant i.e. a single copy (allele) of the mutated gene is enough to cause disease. Dominant negative disorders means that the mutated allele, when expressed, produces large amounts of mutated protein encoded by the mutated allele which negatively competes or interferes with the function or activity of the wild type protein expressed from the wild type allele. This is most commonly observed for proteins that form homomeric complexes, in which the mutant subunits can effectively “poison” the assembly. In some cases the mutation which causes such disorders may be inherited, in others it may develop spontaneously. Dominant gain-of-function disorder means that the mutated allele, when expressed, produces a protein with a different or enhanced activity compared to the wild type protein expressed from a wild type allele. Most dominant negative or gain-of-function disorders are caused by genetic mutations in a gene which is located on a non-sex chromosome and are thereby termed ‘autosomal’.

[0004] In the field of cardiology, dominant negative genetic disorders such as hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), or restrictive cardiomyopathy (RCM) affect large numbers of patients. Often those patients which have more serious forms of the disease are those having dominant negative mutations. For example, TNNI3 mutations accountfor 2.2% total HCM cases in diagnostic labs, and also 0.5% of DCM. The % of HCM patients having such mutations is higher in severe disease setting because TNNI3 often causes restrictive cardiomyopathy / restrictive HCM with uniformly bad outcome. Similarly, MYH7 mutations account for ~15% total HCM and ~5% of DCM cases, and TNNT2 mutations account for ~2% total HCM and ~3 % of DCM cases in diagnostic labs. These patients represent an unmet clinical need.

[0005] Current strategies for using nucleic acid molecules capable of allele-specific binding for treating genetic disorders function through silencing a dominant negative or gain-of-function allele and have focused on targeting and binding to common single nucleotide polymorphisms (SNPs) associated with the pathogenic variant causing the disorder. Most published work in this field relates to single nucleotide substitutions, mostly synonymous coding SNPs. However for treating disorders caused by a heterozygous dominant allele, a very high degree of allele specificity is required. For many disorders, any silencing of the wild-type allele would cause a worsening of the disorder or even be fatal. Achieving complete allelic specificity on the basis of a single nucleotide difference is difficult. Oligonucleotides targeting SNPs are likely to have at least some off-target binding to an allele associated with the wild type copy of the gene. For this reason, current therapeutic nucleic acid molecules have drawbacks for the treatment of such dominant negative or gain-of-function genetic disorders, and patients suffering with these disorders remain without suitable therapies.

[0006] There is a need for nucleic acid therapies which are allele-specific, in particular which only bind to an allele associated with the dominant genetic disorder and which do not affect the expression of the wild type allele, in order to expand the applicability of such therapies to safely treat more genetic disorders.

[0007] One or more aspects or embodiments of the present invention intend to solve the above-mentioned problems in the art. STATEMENTS OF INVENTION

[0008] According to a first aspect of the present disclosure there is provided a therapeutic nucleic acid capable of binding to a target indel allele associated with a pathogenic allele, wherein the pathogenic allele causes a dominant negative genetic disorder or gain-of function genetic disorder, and wherein the disorder is not Huntington’s disease. Advantageously, indelscan be targeted with a high degree of specificity and typically with a higher degree of specificity than SNPs.

[0009] According to a second aspect of the present disclosure there is provided a conjugate comprising the therapeutic nucleic acid of the first aspect covalently linked to a delivery group.

[0010] According to a third aspect of the present disclosure there is provided a delivery particle comprising the therapeutic nucleic acid of the first aspect.

[0011] According to a fourth aspect of the present disclosure there is provided a vector comprising or encoding the therapeutic nucleic acid of the first aspect.

[0012] According to a fifth aspect of the present disclosure there is provided a cell comprising the therapeutic nucleic acid of the first aspect, the conjugate of the second aspect, the delivery particle of the third aspect, or the vector of the fourth aspect.

[0013] According to a sixth aspect of the present disclosure there is provided a pharmaceutical composition comprising the therapeutic nucleic acid according to the first aspect, the conjugate according to the second aspect, the delivery particle of the third aspect, the vector of the fourth aspect, or the cell of the fifth aspect.

[0014] According to a seventh aspect of the present disclosure there is provided a therapeutic nucleic acid according to the first aspect, a conjugate according to the second aspect, a delivery particle acceding to the third aspect, a vector according to the fourth aspect, a cell according to the fifth aspect or a pharmaceutical composition according to the sixth aspect, for use in the treatment or prevention of a dominant negative genetic disorder or gain-of-function genetic disorder, wherein the disorder is not Huntington’s disease.

[0015] According to a eighth aspect of the present disclosure there is provided a method of treatment of a subject having or suspected of having a dominant negative genetic disorder or gain-of-function genetic disorder, or a method of prevention of a dominant negative genetic disorder or gain-of-function genetic disorder in a subject, the method comprising: administering an effective amount of the therapeutic nucleic acid according to the first aspect, a conjugate according to the second aspect, a delivery particle acceding to the third aspect, a vector according to the fourth aspect, a cell according to the fifth aspect, or a pharmaceutical composition according to the sixth aspect, to the subject, wherein the disorder is not Huntington’s disease.

[0016] According to a ninth aspect of the present disclosure there is provided a therapeutic nucleic acid which is capable of binding to a target indel allele of an intronic indel selected from: rs59464879, rs751205475 and rs78373442 associated with a pathogenic HTT allele which causes Huntington’s disease.

[0017] According to a tenth aspect of the present disclosure there is provided a conjugate comprising the therapeutic nucleic acid of the ninth aspect covalently linked to a delivery group.

[0018] According to a eleventh aspect of the present disclosure there is provided a delivery particle comprising the therapeutic nucleic acid of the ninth aspect.

[0019] According to a twelfth aspect of the present disclosure there is provided a vector comprising or encoding the therapeutic nucleic acid of the ninth aspect.

[0020] According to a thirteenth aspect of the present disclosure there is provided a cell comprising the therapeutic nucleic acid of the ninth aspect, the conjugate of the tenth aspect, the delivery particle of the eleventh aspect, or the vector of the twelfth aspect.

[0021] According to a fourteenth aspect of the present disclosure there is provided a pharmaceutical composition comprising the therapeutic nucleic acid according to the ninth aspect, the conjugate according to the tenth aspect, the delivery particle of the eleventh aspect, the vector of the twelfth aspect, or the cell of the thirteenth aspect.

[0022] According to a thirteenth aspect of the present disclosure there is provided a therapeutic nucleic acid according to the ninth aspect, the conjugate according to the tenth aspect, the delivery particle of the eleventh aspect, the vector of the twelfth aspect, the cell of the thirteenth aspect, or a pharmaceutical composition according to the fourteenth aspect, for use in the treatment or prevention of Huntington’s disease.

[0023] According to a fifteenth aspect of the present disclosure there is provided a method of treatment of a subject having or suspected of having Huntington’s disease, or a method of prevention of Huntington’s disease in a subject, the method comprising: administering an effective amount of the therapeutic nucleic acid according to the ninth aspect, the conjugate according to the tenth aspect, the delivery particle of the eleventh aspect, the vector of the twelfth aspect, the cell of the thirteenth aspect, or a pharmaceutical composition according to the fourteenth aspect, to the subject. DEFINITIONS

[0024] As used herein in the present disclosure, unless otherwise clear from context, (i) the term “a” or “an” may be understood to mean “at least one”; (ii) the term “or” may be understood to mean “and / or”; (iii) the terms “comprising”, “comprise”, “including” (whether used with “not limited to” or not), and “include” (whether used with “not limited to” or not) may be understood to encompass itemized components or steps whether presented by themselves or together with one or more additional components or steps; (iv) the term “another” may be understood to mean at least an additional / second one or more; and (v) where ranges are provided, endpoints are included.

[0025] Unless otherwise specified, description of oligonucleotides and elements thereof (e.g., base sequence, sugar modifications, internucleotidic linkages, linkage phosphorus stereochemistry, etc.) is from 5’ to 3’. Unless otherwise specified, oligonucleotides described herein may be provided and / or utilized in salt forms, particularly pharmaceutically acceptable salt forms. As those skilled in the art will appreciate after reading the present disclosure, in some embodiments, oligonucleotides may be provided as salts, but are not limited to, e.g., sodium or potassium salts. As those skilled in the art will appreciate, in some embodiments, individual oligonucleotides within a composition may be considered to be of the same constitution and / or structure even though, within such composition (e.g., a liquid composition), particular such oligonucleotides might be in different salt form(s) (and may be dissolved and the oligonucleotide chain may exist as an anion form when, e.g., in a liquid composition) at a particular moment in time. For example, those skilled in the art will appreciate that, at a given pH, individual internucleotidic linkages along an oligonucleotide chain may be in an acid (H) form, or in one of a plurality of possible salt forms (e.g., a sodium salt, or a salt of a different cation, depending on which ions might be present in the preparation or composition)), and will understand that, so long as their acid forms (e.g., replacing all cations, if any, with H) are of the same constitution and / or structure, such individual oligonucleotides may properly be considered to be of the same constitution and / or structure.

[0026] Nucleic acid: The term “nucleic acid”, as used herein, includes any nucleotides and polymers thereof. The term “polynucleotide”, as used herein, refers to a polymeric form of nucleotides of any length, either ribonucleotides (RNA) or deoxyribonucleotides (DNA) or a combination thereof. These terms refer to the primary structure of the molecules and, thus, include double- and single-stranded DNA, and double- and single-stranded RNA. These terms include, as equivalents, analogs of either RNA or DNA comprising modified nucleotides and / ormodified polynucleotides, such as, though not limited to, methylated, protected and / or capped nucleotides or polynucleotides. The terms encompass poly- or oligo- ribonucleotides (RNA) and poly- or oligo-deoxyribonucleotides (DNA); RNA or DNA derived from N- glycosides or C- glycosides of nucleobases and / or modified nucleobases; nucleic acids derived from sugars and / or modified sugars; and nucleic acids derived from phosphate bridges and / or modified internucleotidic linkages. The term encompasses nucleic acids containing any combinations of nucleobases, modified nucleobases, sugars, modified sugars, phosphate bridges or modified internucleotidic linkages. Examples include, and are not limited to, nucleic acids containing ribose moieties, nucleic acids containing deoxy- ribose moieties, nucleic acids containing both ribose and deoxyribose moieties, nucleic acids containing ribose and modified ribose moieties. Unless otherwise specified, the prefix poly- refers to a nucleic acid containing 2 to about 10,000 nucleotide monomer units and wherein the prefix oligo- refers to a nucleic acid containing 2 to about 200 nucleotide monomer units.

[0027] Nucleotide: The term “nucleotide” as used herein refers to a monomeric unit of a polynucleotide that consists of a nucleobase, a sugar, and one or more internucleotidic linkages (e.g., phosphate linkages in natural DNA and RNA). The naturally occurring bases [guanine, (G), adenine, (A), cytosine, (C), thymine, (T), and uracil (U)] are derivatives of purine or pyrimidine, though it should be understood that naturally and non-naturally occurring base analogs are also included. The naturally occurring sugar is the pentose (five-carbon sugar) deoxyribose (which forms DNA) or ribose (which forms RNA), though it should be understood that naturally and non-naturally occurring sugar analogs are also included. Nucleotides are linked via internucleotidic linkages to form nucleic acids, or polynucleotides. Many internucleotidic linkages are known in the art (such as, though not limited to, phosphate, phosphorothioates, boranophosphates and the like). Artificial nucleic acids include PNAs (peptide nucleic acids), phosphotriesters, phosphorothionates, H-phosphonates, phosphoramidates, boranophosphates, methylphosphonates, phosphonoacetates, thiophosphonoacetates and other variants of the phosphate backbone of native nucleic acids, such as those described herein. In some embodiments, a natural nucleotide comprises a naturally occurring base, sugar and internucleotidic linkage. As used herein, the term “nucleotide” also encompasses structural analogs used in lieu of natural or naturally-occurring nucleotides, such as modified nucleotides and nucleotide analogs. In some embodiments, a “nucleotide” refers to a nucleotide unit in an oligonucleotide or a nucleic acid.

[0028] Oligonucleotide: The term "oligonucleotide" refers to a polymer or oligomer of nucleotides, and may contain any combination of natural and non-natural nucleobases, sugars, and internucleotidic linkages. Oligonucleotides can be single-stranded or double-stranded. A single-stranded oligonucleotide can have double-stranded regions (formed by two portions of the single-stranded oligonucleotide) and a double-stranded oligonucleotide, which comprises two oligonucleotide chains, can have single-stranded regions for example, at regions where the two oligonucleotide chains are not complementary to each other. Example oligonucleotides include, but are not limited to structural genes, genes including control and termination regions, self-replicating systems such as viral or plasmid DNA, single-stranded and double-stranded RNAi agents and other RNA interference reagents (RNAi agents or iRNA agents), shRNA, antisense oligonucleotides, ribozymes, microRNAs, microRNA mimics, supermirs, aptamers, antimirs, antagomirs, Ul adaptors, triplex-forming oligonucleotides, G-quadruplex oligonucleotides, RNA activators, immuno-stimulatory oligonucleotides, and decoy oligonucleotides.

[0029] Oligonucleotides of the present disclosure can be of various lengths. In particular embodiments, oligonucleotides can range from about 2 to about 200 nucleosides in length. In various related embodiments, oligonucleotides, single-stranded, double-stranded, or triple- stranded, can range in length from about 4 to about 10 nucleosides, from about 10 to about 50 nucleosides, from about 20 to about 50 nucleosides, from about 15 to about 30 nucleosides, from about 20 to about 30 nucleosides in length. In some embodiments, the oligonucleotide is from about 15 to about 30 nucleosides in length. In some embodiments, the oligonucleotide is at least 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleosides in length. In some embodiments, the oligonucleotide is at least 4 nucleosides in length. In some embodiments, the oligonucleotide is at least 5 nucleosides in length. In some embodiments, the oligonucleotide is at least 6 nucleosides in length. In some embodiments, the oligonucleotide is at least 7 nucleosides in length. In some embodiments, the oligonucleotide is at least 8 nucleosides in length. In some embodiments, the oligonucleotide is at least 9 nucleosides in length. In some embodiments, the oligonucleotide is at least 10 nucleosides in length. In some embodiments, the oligonucleotide is at least 11 nucleosides in length. In some embodiments, the oligonucleotide is at least 12 nucleosides in length. In some embodiments, the oligonucleotide is at least 15 nucleosides in length. In some embodiments, the oligonucleotide is at least 15 nucleosides in length. In some embodiments, the oligonucleotide is at least 16 nucleosides inlength. In some embodiments, the oligonucleotide is at least 17 nucleosides in length. In some embodiments, the oligonucleotide is at least 18 nucleosides in length. In some embodiments, the oligonucleotide is at least 19 nucleosides in length. In some embodiments, the oligonucleotide is at least 20 nucleosides in length. In some embodiments, the oligonucleotide is at least 25 nucleosides in length. In some embodiments, the oligonucleotide is at least 30 nucleosides in length. In some embodiments, the oligonucleotide is a duplex of complementary strands of at least 18 nucleosides in length. In some embodiments, the oligonucleotide is a duplex of complementary strands of at least 21 nucleosides in length. In some embodiments, each nucleoside counted in an oligonucleotide length independently comprises A, T, C, G, I or U, or optionally substituted A, T, C, G, I or U, or an optionally substituted tautomer of A, T, C, G, I or U. The optionally substituted nucleosides may include 8-oxoA, 8-oxo-dA, 8-oxoT, 8-oxo-dT, 8-oxoC, 8-oxo-dC, 8-oxoG, 8-oxo-dG, 8-oxo-I, 8-oxo-dI, 8-oxoU, and 8-oxo-dU.

[0030] As used herein, the terms “about” and “approximately” may be understood to permit standard variation as would be understood by those of ordinary skill in the art. In some embodiments, the terms “approximately” or “about” in reference to a number are generally taken to include numbers that fall within a range of 5%, 10%, 15%, 20%, 25%, or 30%, in either direction (greater than or less than) of the number unless otherwise stated or otherwise evident from the context.

[0031] Pharmaceutical composition: As used herein, the term “pharmaceutical composition” refers to an active agent, formulated together with one or more pharmaceutically acceptable carriers. In some embodiments, active agent is present in a unit dose amount appropriate for administration in a therapeutic regimen that shows a statistically significant probability of achieving a controlled therapeutic effect when administered to a relevant population.

[0032] Pharmaceutically acceptable: As used herein, the phrase “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0033] Pharmaceutically acceptable salt: The term “pharmaceutically acceptable salt”, as used herein, refers to salts of such compounds that are appropriate for use in pharmaceutical contexts, i.e., salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergicresponse and the like, and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al. describes pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). In some embodiments, a provided compound comprises more than one acid groups, for example, a provided oligonucleotide may comprise two or more acidic groups (e.g., in natural phosphate linkages and / or modified internucleotidic linkages). In some embodiments, a pharmaceutically acceptable salt, or generally a salt, of such a compound comprises two or more cations, which can be the same or different. In some embodiments, in a pharmaceutically acceptable salt (or generally, a salt), each acidic group having sufficient acidity independently exists as its salt form (e.g., in an oligonucleotide comprising natural phosphate linkages and phosphorothioate internucleotidic linkages, each of the natural phosphate linkages and phosphorothioate internucleotidic linkages independently exists as its salt form). In some embodiments, a pharmaceutically acceptable salt of an oligonucleotide is a sodium salt of a provided oligonucleotide. In some embodiments, a pharmaceutically acceptable salt of an oligonucleotide is a sodium salt of a provided oligonucleotide, wherein each acidic linkage, e.g., each natural phosphate linkage and phosphorothioate internucleotidic linkage, exists as a sodium salt form (all sodium salt).

[0034] Subject: As used herein, the term “subject” or “test subject” refers to any organism to which a provided compound or composition is administered in accordance with the present disclosure e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. In some embodiments, a subject may be suffering from, and / or susceptible to a disease, disorder, and / or condition.

[0035] Substantially: As used herein, the term “substantially” refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and / or proceed to completeness or achieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and / or chemical phenomena.

[0036] Suffering from. An individual who is “suffering from” a disease, disorder, and / or condition has been diagnosed with and / or displays one or more symptoms of a disease, disorder, and / or condition.

[0037] Therapeutic agent: As used herein, the phrase “therapeutic agent” refers to any agent that, when administered to a subject, has a therapeutic effect and / or elicits a desired biological and / or pharmacological effect. In some embodiments, a therapeutic agent is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition.

[0038] Therapeutically effective amount: In some embodiments, the term “therapeutically effective amount” means an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that elicits a desired biological response when administered as part of a therapeutic regimen. In some embodiments, a therapeutically effective amount of a substance is an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of the disease, disorder, and / or condition. As will be appreciated by those of ordinary skill in this art, the effective amount of a substance may vary depending on such factors as the desired biological endpoint, the substance to be delivered, the target cell or tissue, etc. For example, the effective amount of compound in a formulation to treat a disease, disorder, and / or condition is the amount that alleviates, ameliorates, relieves, inhibits, prevents, delays onset of, reduces severity of and / or reduces incidence of one or more symptoms or features of the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount, in some embodiments, a single dose is an infusion, which may take up to one or more hours.

[0039] Treat: As used herein, the term “treat,” “treatment,” or “treating” refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of, and / or reduce incidence of one or more symptoms or features of a disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject who exhibits only early signs of the disease, disorder, and / or condition, for example for the purpose of decreasing the risk of developing pathology associated with the disease, disorder, and / or condition.

[0040] Identity: As used herein, the term “identity” refers to the overall relatedness between polymeric molecules, e.g., between nucleic acid molecules (e.g., oligonucleotides, DNA, RNA,etc.) and / or between polypeptide molecules. In some embodiments, polymeric molecules are considered to be “substantially identical” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical. Calculation of the percent identity of two nucleic acid or polypeptide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or substantially 100% of the length of a reference sequence. The nucleotides at corresponding positions are then compared. When a position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller (CABIOS, 1989, 4: 11-17), which has been incorporated into the ALIGN program (version 2.0). In some exemplary embodiments, nucleic acid sequence comparisons made with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna.CMP matrix.

[0041] Wild-type: As used herein, the term “wild-type” has its art-understood meaning that refers to an entity having a structure and / or activity as found in nature in a “normal” (as contrasted with mutant, diseased, altered, etc.) state or context. Those of ordinary skill in the art will appreciate that wild type sequences, genes and polypeptides often exist in multiple different forms (e.g., alleles). In the case of dominant negative genetic disorders or gain-of-function genetic disorders, a subject may have one copy of a pathogenic allele, and one copy of a wild type allele of the same gene.

[0042] Further features and embodiments of the present invention will now be described under the following headed sections. Any feature in any section may be combined with any embodiment or aspect herein in any workable combination. DETAILED DESCRIPTION

[0043] Therapeutic nucleic acid

[0044] The therapeutic nucleic acid of the invention targets and binds to an indel allele associated with a disease-causing (i.e. pathogenic) allele which causes a dominant negative genetic disorder or a dominant gain-of-function genetic disorder.

[0045] Suitably, the therapeutic nucleic acid is at least partially complementary to the target indel allele associated with a disease-causing (pathogenic) allele. Suitable indel alleles associated with disease-causing (pathogenic) alleles are described below.

[0046] Suitably, the therapeutic nucleic acid comprises a target recognition sequence. The target recognition sequence is a region of complementarity to the target nucleic acid, typically sufficiently complementary to the target nucleic acid to allow hybridisation under physiological conditions. The target recognition sequence may specifically hybridise to the target nucleic acid.

[0047] Suitably the target nucleic acid is a nucleic acid molecule comprising the target indel allele. Suitably therefore the therapeutic nucleic acid targets and binds to a target nucleic acid comprising a target indel allele. Suitably therefore the therapeutic nucleic acid comprises a target recognition sequence which targets and binds to a target nucleic acid comprising a target indel allele.

[0048] Suitably the target nucleic acid is a transcript, suitably the target nucleic acid is RNA, suitably the target nucleic acid is pre-mRNA or mRNA. In one embodiment, the target nucleic acid is pre-mRNA. Suitably therefore the target nucleic acid is pre-mRNA comprising the target indel allele. Suitably therefore, the therapeutic nucleic acid of the invention comprises a target recognition sequence capable of binding to a target nucleic acid sequence, wherein the target nucleic acid sequence comprises a target indel allele associated with a disease-causing (pathogenic) allele, wherein the pathogenic allele causes a dominant negative genetic disorder or a dominant gain-of-function genetic disorder.

[0049] In one embodiment, the therapeutic nucleic acid of the invention is capable of binding to a target indel allele present within pre-mRNA encoding a disease-causing (pathogenic) allele, wherein the pathogenic allele causes a dominant negative genetic disorder or a dominant gain-of-function genetic disorder. Suitably therefore, the therapeutic nucleic acid of the invention comprises a target recognition sequence capable of binding to a target indel allele present within pre-mRNA encoding a disease-causing (pathogenic) allele, wherein the pathogenic allele causes a dominant negative genetic disorder or a dominant gain-of-function genetic disorder. Suitably therefore, the therapeutic nucleic acid of the invention comprises a target recognition sequence capable of binding to a target nucleic acid sequence, wherein the target nucleic acid sequence comprises a target indel allele present within pre-mRNA encoding a disease-causing (pathogenic) allele, wherein the pathogenic allele causes a dominant negative genetic disorder or a dominant gain-of-function genetic disorder.

[0050] Suitably the target recognition sequence may be up to 50, 40, 30, 20, 10 or 5 nucleotides in length. The target recognition sequence may be at least 15, 20, 25, 30 or 35 nucleotides in length. For example, the target recognition sequence may be between 15 to 25 nucleotides, or between 18 to 25 nucleotides in length. In a particular embodiment, the target recognition sequence is 16 nucleotides in length. The length may vary from target to target but may be routinely determined by a person having ordinary skill in the art. Suitably the target recognition sequence comprises a length which is suitable to bind to the target nucleic acid sequence, suitably a length with is suitable to bind to the target indel allele, suitably to the entire target indel allele. Suitably the target nucleic acid sequence may have approximately the same length as the target recognition sequence. Suitably the target nucleic acid sequence comprises at least the target indel allele, and optionally one or more additional nucleotides which may surround the target indel allele, suitably at the 5' and 3' ends thereof. Suitably the target nucleic acid sequence has a length which is suitable to encompass the target indel allele.

[0051] The target recognition sequence may be partially or fully complementary to the target nucleic acid. The target recognition sequence may comprise one or more (e.g. ≤5, ≤4, ≤3, ≤2 or 1) mismatch nucleobases when aligned with the target nucleic acid. Suitably, the target recognition sequence has a complementarity of over 75%, over 80%, over 85%, over 90%, over 95%, over 96%, over 97%, over 98%, over 99%, or up to 100% complementarity with the target nucleic acid sequence.

[0052] The target recognition sequence may comprise ≥2, ≥3, ≥4, ≥5, ≥6, ≥7, ≥8, ≥9, ≥10, ≥11, ≥12, ≥13, ≥14, ≥15, ≥16, ≥17, ≥18, ≥19, and / or ≥20 contiguous nucleotides that are complementary to their opposing nucleotides in the target nucleic acid.

[0053] Suitably, the target recognition sequence has complementarity to the target nucleic acid sequence across at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% of its total length.

[0054] It is understood that in order for the therapeutic nucleic acid to be capable of binding, it does not require that the entire length of the therapeutic nucleic acid binds to the target nucleic acid. It will be appreciated that a portion of the therapeutic nucleic acid may not bind to the target nucleic acid, for example the 5' or the 3' ends of the therapeutic nucleic acid.

[0055] Suitably the therapeutic nucleic acid may have a total length of between 10 to 60 nucleotides, suitably between 10 to 50, 10 to 40, 10 to 30 nucleotides. In one embodiment, the total length of the therapeutic nucleic acid is between 15 to 30 nucleotides in length. In one embodiment, the therapeutic nucleic acid is at least 15 nucleotides in length. Suitably the therapeutic nucleic acid comprises the target recognition sequence. Suitably therefore the therapeutic nucleic acid has a length which at least encompasses the target recognition sequence, and optionally one or more additional nucleotides suitably at the 5' and 3' ends thereof. In one embodiment, the total length of the therapeutic nucleic acid consists of the target recognition sequence.

[0056] Suitably the target recognition sequence is positioned approximately in the centre of the entire therapeutic nucleic acid. Suitably the target recognition sequence comprised in the therapeutic nucleic acid begins at between the 5thto the 10thnucleotide in the therapeutic nucleic acid (from the 5’ end), in one embodiment the therapeutic nucleic acid begins at around the 8thnucleotide in the therapeutic nucleic acid (from the 5’ end).

[0057] Suitably, the therapeutic nucleic acid targets and binds to the target indel allele associated with a disease-causing (pathogenic) allele, and / or reduces the level, expression and / or activity of the pathogenic allele or transcripts thereof, with a level of allelic discrimination of at least 1.1-fold, suitably at least 1.2-fold, suitably at least 1.3-fold, suitably at least 1.4-fold, suitably at least 1.5-fold, suitably at least 1.6-fold, suitably at least 1.7-fold, suitably at least 1.8-fold, suitably at least 1.9-fold, suitably at least 2-fold, suitably at least 3-fold, suitably at least 4-fold, suitably at least 5-fold, suitably at least 10-fold. In some embodiments, the level of allelic discrimination of at least 1.3-fold. In some embodiments, the level of allelic discrimination of at least 1.5-fold. In some embodiments, the level of allelic discrimination of at least 2.0-fold. Suitably, the level of allelic discrimination may be determined by using an assay to determine the level of binding of the therapeutic nucleic acid to either form of an indeland dividing the result of the level of binding to the on-target indel by the level of binding to off-target indel. Exemplary assays for determining allelic discrimination are provided in the Examples. For example, the therapeutic nucleic acid may reduce the level, expression and / or activity of an allele associated with an on-target indel at least 2-fold more than the level, expression and / or activity of an allele associated with an off-target indel as measured using, for example, the dual luciferase reporter assay described in Example 3.

[0058] Suitably the therapeutic nucleic acid does not bind to any sequence associated with the corresponding wild type allele of the disease-causing (pathogenic) allele. Suitably the therapeutic nucleic acid does not bind to the alternative indel allele which is associated with the corresponding wild type allele of the disease-causing (pathogenic) allele.

[0059] Suitably, the therapeutic nucleic acid may be selected from any type of known therapeutic nucleic acid such as: ASO, gRNA, siRNA, shRNA, miRNA etc. Suitably, the therapeutic nucleic acid is an oligonucleotide. In one embodiment, the therapeutic nucleic acid is an antisense oligonucleotide (ASO).

[0060] Suitably the ASO may comprise any oligonucleotide. An oligonucleotide is an oligomer comprising two or more nucleotides. The oligonucleotide may comprise DNA or RNA. The nucleotides can be naturally occurring or artificial. A nucleotide typically contains a nucleobase, a sugar and at least one linking group, such as a phosphate, 2’O-methyl, 2’ methoxy- ethyl, phosphoramidate, methylphosphonate or phosphorothioate group. The nucleobase is typically heterocyclic. Nucleobases include, but are not limited to, purines and pyrimidines and more specifically adenine (A), guanine (G), thymine (T), uracil (U) and cytosine (C). The sugar is typically a pentose sugar. Nucleotide sugars include, but are not limited to, ribose and deoxyribose. The nucleotide is typically a ribonucleotide or deoxyribonucleotide. The nucleotide typically contains a monophosphate, diphosphate or triphosphate. Phosphates may be attached on the 5’ or 3’ side of a nucleotide.

[0061] In some embodiments, the oligonucleotide comprises 2’ O-methoxyethyl modifications (MOE), peptide nucleic acid modifications (PNA), bridged nucleic acid modifications (BNA), locked nucleic acid modifications (LNA) and / or phosphorodiamidate morpholino oligomers (PMO). The nucleotides may contain additional modifications. In particular, suitable modified nucleotides include, but are not limited to, 2’amino pyrimidines (such as 2’-amino cytidine and 2’-amino uridine), 2’-hydroxyl purines (such as , 2’-fluoro pyrimidines (such as 2’-fluorocytidine and 2’fluoro uridine), hydroxyl pyrimidines (such as 5’-α-P-borano uridine), 2’-O-methyl nucleotides (such as 2’-O-methyl adenosine, 2’-O-methyl guanosine, 2’-O-methyl cytidine and 2’-O-methyl uridine), 4’-thio pyrimidines (such as 4’-thio uridine and 4’-thio cytidine) and nucleotides have modifications of the nucleobase (such as 5- pentynyl-2’-deoxy uridine, 5-(3-aminopropyl)-uridine and 1,6-diaminohexyl-N-5- carbamoylmethyl uridine). In one embodiment, the oligonucleotide comprises 2’ O- methoxyethyl modifications (MOE) and / or locked nucleic acid modifications (LNA).

[0062] The oligonucleotide may comprise 3ʹ to 5ʹ phosphodiester (PO) linkages as naturally found in DNA or RNA. The oligonucleotide may comprise modified internucleoside linkages, e.g. phosphotriester, phosphorothioate (PS), boranophosphate, phosphorodiamidate, phosphoamidate, and / or thiophosphoramidate linkages. The modified internucleoside linkage may be selected from any other modifications known in the art. In one embodiment, the oligonucleotide comprises phosphorothioate (PS) linkages. In some embodiments, the ASO comprises bridged nucleic acids. In some embodiments, the ASO comprises locked nucleic acids.

[0063] The oligonucleotide may comprise one or more asymmetric centres and thus give rise to enantiomers, diastereomers, and other stereoisomeric configurations, e.g. R, S. For example, stereochemistry at the phosphorus atom in the internucleoside linkage can be manipulated to produce phosphorothioate (PS) linkages with controlled R and S configurations. Stereochemistry may be constrained at one or more modified internucleoside linkages. This stereochemical precision can reduce off-target effects and improve the pharmacokinetic and pharmacodynamic profiles of the oligonucleotides. Suitably the oligonucleotide is at least about 10%, 20%, 30%, 40%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% diastereomerically pure.

[0064] Suitably the oligonucleotide may comprise one or more chiral internucleotide linkages. The use of chiral internucleotidic linkages in oligonucleotide synthesis can create a chiral center at the linkage phosphorus. A chiral linkage phosphorus center can have either an “Sp” or “Rp” configuration. A conventional stereorandom preparation of an oligonucleotide that contains n chiral linkage phosphorus is a mixture of 2nstereoisomers with respect to chiral linkage phosphorus centers, each of which stereoisomers share the same constitution but differs in stereochemistry along its backbone. Without control of stereochemistry of backbone chiral centers, a stereorandom oligonucleotide preparation (e.g., a random mixture of diastereoisomers) provides uncontrolled (or stereorandom) compositions comprising undetermined levels ofoligonucleotide stereoisomers. Even though these stereoisomers may have the same base sequence and / or chemical modifications, they are different chemical entities at least about due to their different backbone stereochemistry, and they can have different properties, e.g., activities, toxicities, distribution etc. In some embodiments, each chiral internucleotidic linkage is independently formed with about 97% or more diastereoselectivity (e g., as measured through preparation of a suitable dimer). In some embodiments, most chiral intemucleotidic linkages are independently formed with about 98% or more diastereoselectivity. In some embodiments, one or more, e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more chiral internucleotidic linkages are independently formed with about 99% or more diastereoselectivity. In some embodiments, overall diastereoselectivity (as product of diastereoselectivity of all chiral internucleotidic linkages) is about 80% or more. In some embodiments, it is about 81% or more. In some embodiments, it is about 82% or more. In some embodiments, it is about 83% or more. In some embodiments, it is about 84% or more. In some embodiments, it is about 85% or more. In some embodiments, the oligonucleotide is stereodefined or stereopure. In some embodiments, the oligonucleotide may therefore be referred to as chirally controlled. In some embodiments, level of a particular stereoisomer, of a chirally controlled oligonucleotide composition is enriched as described herein (e.g., in some embodiments, each chiral internucleotidic linkage independently has a stereopurity of about 97%, 98%, 99% or more). In some embodiments, each chirally controlled internucleotide linkage independently has a diastereopurity of at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% with respect to its chiral linkage phosphorus.

[0065] The oligonucleotide may comprise a single type of nucleic acid chemistry as described herein or combinations of different nucleic acid chemistries.

[0066] For example, each of the sugar moieties in the oligonucleotide may comprise a 2ʹ-O- methoxyethyl (2ʹ-MOE) modification and each of the internucleoside linkages may be a phosphorothioate (i.e. a fully PS-MOE oligonucleotide). PS modifications are known to result in resistance to a broad spectrum of nucleases and increase protein binding, which also improves tissue uptake.2ʹ-MOE modifications are known to enable enhanced binding affinity to the target pre-mRNA with minimal toxicity and reduce plasma protein binding.

[0067] Suitably, the oligonucleotide may comprise a combination of PO and PS internucleoside linkages. This may facilitate the fining tuning of the pharmacokinetics of the oligonucleotide.

[0068] In some embodiments, the antisense oligonucleotide is a gapmer antisense oligonucleotide i.e. ‘a gapmer’. A gapmer ASO comprises a central region or ‘gap’ flanked on either side by a first and a second flanking region. Suitably the central region comprises a DNA- based internal ‘gap’ which is unmodified or which comprises 2ʹ-O-methyl (2ʹ-OMe) modifications. Suitably the two flanking regions comprise RNA, suitably modified RNA, often comprising for example: 2ʹ-O-methyl (2ʹ-OMe), locked nucleic acid (LNA), S-cEt oligonucleotides (constrained ethyl nucleotides), tricycle-DNA, or 2ʹ-O-methoxyethyl (2ʹ- MOE) modifications, suitably that bind to targets with high affinity.

[0069] In some embodiments, the structure of an oligonucleotide comprises or consists of an asymmetrical format. In some embodiments, the structure of an oligonucleotide comprises or consists of a symmetrical format. Suitably therefore in some embodiments the gapmer oligonucleotide may comprise flanking regions of different lengths, and comprising different modifications.

[0070] In one embodiment, the antisense oligonucleotide is a gapmer comprising a symmetrical format.

[0071] Suitably the gapmer comprises a first flanking region and a second flanking region each comprising modified nucleotides, and a central region consisting of unmodified nucleotides. Suitably the gapmer comprises between 2-10 nucleotides in each flanking region, suitably between 3-5 nucleotides in each flanking region. In some embodiments, 5 nucleotides in each flanking region. Suitably wherein one or more of the nucleotides in each flanking region is modified, suitably wherein the majority of nucleotides in each flanking region is modified, suitably wherein each flanking region consists of modified nucleotides.

[0072] Suitably the modified nucleotides are modified RNA nucleotides. Suitably the modified nucleotides are 2ʹ-O-methoxyethyl (2ʹ-MOE) modified nucleotides and / or locked nucleotides. Suitably wherein the 2’-MOE modified nucleotides may be indicated as MOE- nucleotide, wherein ‘-‘ indicates the attachment of the MOE to the following nucleotide. Suitably each flanking region may comprise only one type of modified nucleotide or may comprise a mixture of differently modified nucleotides. Suitably each flanking region may comprise only 2ʹ-O-methoxyethyl (2ʹ-MOE) modified nucleotides. Alternatively each flanking region may comprise only locked nucleotides. Alternatively still, each flanking region may comprise both 2ʹ-O-methoxyethyl (2ʹ-MOE) modified nucleotides and locked nucleotides.

[0073] Suitably, to allow RNAse H binding to the heteroduplex and cleavage of the target RNA comprising the target indel allele , the central region of the oligonucleotide comprises unmodified nucleotides. Suitably the central region comprises a length of between 7-10 nucleotides.

[0074] In some cases, to reduce cleavage in off-target effects with partial complementarity, modifications in the flanking regions can be distributed asymmetrically, for example the modifications in the first and the second flanking region may be distributed as follows, wherein the first number indicates the number of modified positions in the first flanking region (5’) and the second number indicates the number of modified positions in the second flanking region (3’): (3 / 7), (4 / 6), (5 / 5), (6 / 4), (7 / 3), (3 / 6), (4 / 5), (5 / 4), (6 / 3), (3 / 5), (4 / 4), (5 / 3), (3 / 4), (4 / 3), or (3 / 3).

[0075] In one embodiment, the oligonucleotide is a gapmer comprising a first and a second flanking region and a central region therebetween, wherein the first and second flanking regions consist of 2’MOE modified nucleotides, and wherein the central region consists of unmodified DNA. Suitably wherein the first and second flanking regions comprise between 3-5 nucleotides and the central region comprises between 7-10 nucleotides. In one embodiment wherein the first and second flanking regions consist of 5 nucleotides and the central region consists of 8 nucleotides. Suitably wherein the oligonucleotide comprises phosphorothioate (PS) linkages.

[0076] In one embodiment, the oligonucleotide is a gapmer comprising a first and a second flanking region and a central region therebetween, wherein the first and second flanking regions consist of locked nucleotides, and wherein the central region consists of unmodified DNA. Suitably wherein the first and second flanking regions comprise between 3-5 nucleotides and the central region comprises between 7-10 nucleotides. In one embodiment wherein the first and second flanking regions consist of 3 nucleotides and the central region consists of 10 nucleotides. Suitably wherein the oligonucleotide comprises phosphorothioate (PS) linkages.

[0077] In one embodiment, the oligonucleotide is a gapmer comprising a first and a second flanking region and a central region therebetween, wherein the first and second flanking regions consist of 2’MOE modified nucleotides and locked nucleotides, and wherein the central region consists of unmodified DNA. Suitably wherein the first and second flanking regions comprise between 3-5 nucleotides and the central region comprises between 7-10 nucleotides. In one embodiment wherein the first and second flanking regions consist of 5 nucleotides and the central region consists of 10 nucleotides. In one embodiment wherein each flanking regionconsists of two locked nucleotides and three 2’MOE modified nucleotides. Suitably wherein the oligonucleotide comprises phosphorothioate (PS) linkages.

[0078] Optionally the therapeutic nucleic acid may comprise one or more terminal 5’ or 3’ modifications. Optionally the therapeutic nucleic acid may comprise a phosphate, inverted dT, a lipid such as cholesterol, palmitic acid or oleic acid, polyethylene glycol (PEG) or other polymer, a fluorescent label such as FAM or Cy5, biotin or other affinity tag, a methyl group, a fluorine, or an antibody, cell penetrating peptide, or GalNAc group at the 3’ or the 5’ end thereof, for example.

[0079] Suitably the therapeutic nucleic acid comprises a sequence selected from any one of SEQ ID NOs: 1-32, 33-84, 85-145, 146-187, 232, 188-219, and 260, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, or a sequence which targets and binds to the alternative indel allele at the same indel site to that targeted and bound by SEQ ID NO: 1-32, 33-84, 85-145, 146-187, 232, 188-219 or 260. Suitably the therapeutic nucleic acid comprises a sequence selected from any one of SEQ ID NOs: 1-32, 33-78, 85-139, 146-181, 232, 188-191, 199-202, and 212-215, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, or a sequence which targets and binds to the alternative indel allele at the same indel site to that targeted and bound by SEQ ID NO: 1-32, 33-78, 85-139, 146-181, 232, 188-191, 199-202, and 212-215. Suitably the therapeutic nucleic acid comprises a sequence selected from SEQ ID NO: 66, 87, 101, 156 and 199, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably the therapeutic nucleic acid consists of a sequence selected from SEQ ID NO: 66, 87, 156 and 199.

[0080] In one further aspect of the invention there is provided a therapeutic nucleic acid, optionally an antisense oligonucleotide, comprising a sequence according to any of SEQ ID NOs: 33-78, 85-139, 146-181, 232, 188-191, 199-202, and 212-215, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, or a sequence which targets and binds to the alternative indel allele at the same indel site to thattargeted and bound by SEQ ID NO: 1-32, 33-78, 85-139, 146-181, 232, 188-191, 199-202, and 212-215.

[0081] In one embodiment, the therapeutic nucleic acid comprises a sequence selected from any one of SEQ ID NO: 85-145, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which targets and binds to an indel allele of the intronic indel rs377373012. In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 85-139, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which targets and binds to an indel allele of the intronic indel rs377373012. In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 87 (E3), SEQ ID NO: 92-95 (E8-E11), SEQ ID NO: 101 (E17), SEQ ID NO: 103 (E19), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which targets and binds to an indel allele of the intronic indel rs377373012. In one embodiment, the therapeutic nucleic acid is an ASO and consists of a sequence selected from any one of SEQ ID NO: 87 (E3), SEQ ID NO: 92-95 (E8-E11), SEQ ID NO: 101 (E17), SEQ ID NO: 103 (E19). Suitably which targets and binds to an indel allele of the intronic indel rs377373012. In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 85-107 (E3-E23), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, suitably which targets and binds to an indel allele of the intronic indel rs377373012. In one embodiment, the therapeutic nucleic acid is an ASO and consists of a sequence selected from any one of SEQ ID NO: 85-107 (E3-E23), suitably which targets and binds to an indel allele of the intronic indel rs377373012. In one embodiment, the therapeutic nucleic acid is an ASO and comprises SEQ ID NO: 87 (E3) or SEQ ID NO: 101 (E17), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which targets and binds to an indel allele of the intronic indel rs377373012. In one embodiment, the therapeutic nucleic acid is an ASO and consists ofSEQ ID NO: 87 (E3) or SEQ ID NO: 101 (E17). Suitably which targets and binds to an indel allele of the intronic indel rs377373012. Suitably which targets and binds to an indel allele at intronic indel site rs377373012. Suitably which is associated with the pathogenic allele TNNI3.

[0082] In one embodiment, the therapeutic nucleic acid comprises a sequence selected from any one of SEQ ID NO: 33-84, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which target and bind to an indel allele of the intronic indel rs34598192. In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 33-78, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 60-66 (D1-D7), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which targets and binds to an indel allele of the intronic indel rs34598192. In one embodiment, the therapeutic nucleic acid is an ASO and consists of a sequence selected from any one of SEQ ID NO: 60-66 (D1-D7), suitably which targets and binds to an indel allele of the intronic indel rs34598192. In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 33-37 (C1-C5), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, suitably which targets and binds to an indel allele of the intronic indel rs34598192. In one embodiment, the therapeutic nucleic acid is an ASO and consists of a sequence selected from any one of SEQ ID NO: 33-37 (C1-C5), suitably which targets and binds to an indel allele of the intronic indel rs34598192. Suitably which target and bind to an indel allele of the intronic indel rs34598192. In one embodiment, the therapeutic nucleic acid is an ASO and comprises SEQ ID 66 (D7), SEQ ID NO: 33 (C1), or SEQ ID 36 (C4), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which targets and binds to an indel allele of the intronic indel rs34598192. In one embodiment, the therapeutic nucleic acid is an ASO and consists of SEQ ID 66 (D7), SEQ ID NO: 33 (C1),or SEQ ID 36 (C4). Suitably which targets and binds to an indel allele of the intronic indel rs34598192. Suitably which targets and binds to an indel allele at intronic indel site rs34598192. Suitably which is associated with the pathogenic allele MYH7.

[0083] In one embodiment, the therapeutic nucleic acid comprises a sequence selected from any one of SEQ ID NO: 146-187 and 232, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which target and bind to an indel allele of the intronic indel rs45533739. In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 146- 181 and 232, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which target and bind to an indel allele of the intronic indel rs45533739. In one embodiment, the therapeutic nucleic acid is an ASO and comprises SEQ ID NO: 156, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably which targets and binds to an indel allele of the intronic indel rs45533739. In one embodiment, the therapeutic nucleic acid is an ASO and consists of SEQ ID NO: 156, which targets and binds to an indel allele of the intronic indel rs45533739. Suitably which targets and binds to an indel allele at intronic indel site rs45533739. Suitably which is associated with the pathogenic allele TNNT2.

[0084] Target Indel Allele

[0085] An indel is an insertion or deletion in a nucleic acid sequence, suitably an insertion and / or deletion of one or more nucleotides into genomic DNA of less than 1 kb in length. The therapeutic nucleic acid of the invention is capable of binding to a target indel allele which is associated with a pathogenic allele in the subject to be treated, wherein the pathogenic allele causes a dominant negative or gain-of function genetic disorder. Suitably by ‘indel allele’ in the context of the present invention it is meant a DNA polymorphism, otherwise known as an insertion / deletion polymorphism, which is similar to an SNP but involves an insertion or a deletion of nucleotides rather than simply a difference in a single nucleotide. Essentially, segments of DNA in specific locations or sites in the genome are missing in some individuals of a population and present in others. Suitably an indel allele is characterized by the presence or insertion (one allele), or the absence or deletion (the alternative allele) of a sequence of DNA,at given ‘indel’ site or position, which sequence can vary. Suitably wherein the site is designated with an rsID number. Suitably, the rsID numbers mentioned herein refer to version 156 of the Single Nucleotide Polymorphism Database (dbSNP), which is maintained by the National Center for Biotechnology Information (NCBI) and is accessible via www.ncbi.nlm.nih.gov / snp or via Genome Aggregation Database (gnomAD) v4.1.0 (Chen et al 2024). Whilst many indel sites are biallelic i.e. two different alleles are possible at the indel site, some indel sites may be multi-allelic i.e. have more than two alleles. Therefore, there may be more than one different insertion or deletion allele possible at a given indel site. Suitably an indel allele is therefore either the presence / insertion of a sequence or the absence / deletion of a sequence in the genome of a subject at a particular site / position or locus. Suitably the particular position or locus of an indel in a genome is typically given by an rsID number as used herein or a chromosome number and location as used for example in Figure 21. Suitably wherein in the present invention, a target indel allele (either an insertion or a deletion) at the recited indel site happens to be associated with the pathogenic allele in said subject. What is meant by ‘associated with’ in this context is defined below.

[0086] Suitably by 'capable of binding' it is meant that the therapeutic nucleic acid comprises a sequence which is able to bind to the target indel allele, i.e. Which is at least partially complementary to the target indel allele. In other words, this may be regarded as the therapeutic nucleic acid having the ability to target the indel allele. Typically the therapeutic nucleic acid comprises a target recognition sequence which is complementary to a target nucleic acid region comprising the target indel allele, as described above.

[0087] Suitably by ‘associated with’ it is meant that the target indel allele is on the same haplotype as the pathogenic allele in the subject to be treated. Suitably therefore the indel allele is in cis or in phase with the pathogenic allele. Suitably this may mean that the indel allele is present within the pathogenic allele itself, or may be present within a different sequence which is located close to the pathogenic allele, and is therefore on the same haplotype, or on the same chromosome as the pathogenic allele. Suitably, in any case, the indel allele is expressed with the pathogenic allele. Suitably the target indel allele is present within the same transcript as the pathogenic allele, suitably therefore the target indel allele is present within pre-mRNA or mRNA encoding the pathogenic allele. Suitably the indel allele may be pathogenic i.e. it may be a cause of the pathogenesis of the disease, or a contributing factor to the disease in the subject, howeveroften the indel allele is not in itself pathogenic but is simply associated with the pathogenic allele.

[0088] In some preferred embodiments, the therapeutic nucleic acid targets an indel allele which is located within a pathogenic allele which causes said genetic disorder. Suitably the indel allele is specifically associated with the pathogenic allele which causes said genetic disorder.

[0089] Suitably the target indel allele is not associated with the corresponding wild type allele to the pathogenic allele in the subject. Suitably therefore the target indel allele is not on the same haplotype as the wild type allele in the subject. Suitably therefore the target indel allele is not present in transcripts, i.e. In pre-mRNA or mRNA, encoding the wild type allele. Suitably therefore the therapeutic nucleic acid of the invention is allele-specific.

[0090] Suitably the indel allele may be selected from: an insertion and / or a deletion, and may be of any length of less than 1Knt or 1Kb. Suitably, the insertion may be of between 1 and 50, 1 and 40, 1 and 30, 1 and 20, 1 and 10 nucleotides, or base pairs. Suitably the deletion may be of between 1 and 50, 1 and 40, 1 and 30, 1 and 20, 1 and 10 nucleotides or base pairs.

[0091] Suitably, the target indel allele may be located in a coding or a non-coding region of a nucleic acid, suitably of a target nucleic acid as described above. Suitably the target indel allele is located in a non-coding region such as a UTR or an intron of a nucleic acid, suitably of a target nucleic acid as described above. In one embodiment, the target indel allele is located in an intron of a nucleic acid, suitably of a target nucleic acid as described above. Therefore in one embodiment, the target indel allele is an intronic indel allele.

[0092] Suitably the target indel allele is heterozygous in the subject. Suitably therefore at the genetic locus or site of the target indel allele, the subject having the dominant negative or gain- of function genetic disorder is heterozygous. Suitably both alleles of the indel are common. Suitably therefore the target indel allele has a high minor allele frequency (the frequency at which the second most common allele occurs in a given population, MAF), suitably across different ancestry populations. Preferably the target indel allele has a MAF of at least 0.01, at least 0.06, at least 0.07, at least 0.08, at least 0.09, at least 0.1, suitably between 0.1 and 0.5. Suitably the target indel allele has a MAF of about 0.1, 0.2, 0.3, 0.4, or 0.5.

[0093] Suitably the target indel allele may be any indel listed in Figure 21. Suitably the target indel allele may be any indel listed in column ‘genome variant id’ Figure 21. For example, the target indel may be in the AARS1 gene and may be either C or CA at position 70265687 on chromosome 16 of human genome GRCh38.p14. Suitably therefore, the sequence C or CA atposition 70265687 on chromosome 16 of human genome GRCh38.p14 may be associated with a pathogenic AARS1 allele. Suitably therefore, there is provided a therapeutic nucleic acid capable of binding to C or CA at position 70265687 on chromosome 16 of human genome GRCh38.p14.

[0094] Pathogenic Allele

[0095] The present invention relates to therapeutic nucleic acids for targeting and binding to a target indel allele which is associated with a pathogenic allele in a subject, thereby providing a means to reduce expression of the pathogenic allele, and treatment of the disease in the subject.

[0096] Suitably the pathogenic allele which causes a dominant negative or gain-of function genetic disorder may be selected from any gene which is known to cause a dominant negative or gain-of function genetic disorder. Suitably the pathogenic allele may be selected from any gene mentioned in Figure 21. Suitably the pathogenic allele may be selected from any gene mentioned in column ‘gene id’ or ‘gene symbol’ of Figure 21. Suitably, a pathogenic allele mentioned in column ‘gene id’ or ‘gene symbol’ of Figure 21 may cause or be associated with a disorder listed in column ‘phenotype’ of Figure 21. For example, the pathogenic allele may be AARS1 and may cause Charcot-Marie-Tooth disease. For example, the pathogenic allele may be ABCA4 and may cause macular degeneration. Suitably therefore, the pathogenic allele may be selected from a gene which causes a disorder listed in column ‘phenotype’ of Figure 21.

[0097] Suitably, the pathogenic allele may be selected from any one of the following genes: AARS1, ABCA4, ABCA7, ABCB6, ABCC6, ABCC8, ACOX1, ACTA1, ACTL6B, ACVR1, ADCY10, ADGRE2, ADH1C, ADRB3, AFG3L2, AFP, AGBL1, AGRP, AIPL1, ALDH18A1, ALDH2, ALG10B, ALG5, ALG8, ALOX5, ALPK1, ANGPT2, ANKRD26, ANO1, ANO3, ANO5, ANXA11, ANXA5, APOA4, APOA5, APOE, APOL1, AQP2, AQP5, ARMC5, ASB10, ASPN, ATAD3A, ATL1, ATL3, ATP8B1, ATXN1, ATXN10, ATXN7, AVP, BEAN1, BEST1, BFSP1, BFSP2, BHLHE41, BICRA, BLK, BLVRA, BSCL2, BTNL2, C1S, C9, C9orf72, CACNA1H, CACNA1S, CACNB2, CACNB4, CALCR, CAPN3, CAPN5, CARD14, CASQ1, CAV1, CAV3, CBL, CCL11, CCL2, CCM2, CD46, CD96, CDH15, CDH23, CDON, CDSN, CEACAM16, CEBPE, CEL, CELA2A, CEP85L, CFAP43, CFB, CFHR1, CFHR3, CFI, CFTR, CHEK1, CHI3L1, CHMP2B, CHN1, CHRNA1, CHRNA2, CHRNA4, CHRNB1, CHRNE, CILK1, CLCN1, CLCN2, CLCN6, CLCN7, CLEC3B, CLPB, COCH, COG4, COL10A1, COL17A1, COL18A1, COL1A1, COL1A2, COL2A1, COL4A2, COL4A3, COL4A4, COL6A1, COL6A2, COL7A1, COL8A2, COL9A2, COL9A3, COMP, COMT, COQ2, CORIN, CPA6, CPOX, CPSF1, CRB1,CREB3L3, CRELD1, CRY1, CRYAB, CRYBA1, CRYBA4, CRYBB1, CRYBB2, CRYBB3, CRYGB, CRYGC, CRYGD, CRYGS, CRYM, CSRP3, CST3, CTNNA3, CTRC, CYP11B1, CYP2C9, CYP3A4, DEAF1, DES, DHX16, DIABLO, DIAPH3, DIO1, DMPK, DNA2, DNAAF4, DNAJB11, DNAJB6, DNASE1, DPP6, DRD4, DRD5, DSC2, DSPP, DVL1, EDAR, EDARADD, EDN1, EFHC1, EGLN1, EIF2AK2, ELANE, ELP1, EMILIN1, ENAM, ENPP1, EPB41, EPCAM, EPHA2, EPHB4, EPHX2, EPO, EPOR, ERCC6, ERLIN2, EVC2, F12, F5, FAM83H, FCGR2A, FCGR2B, FDPS, FGA, FGB, FGF23, FGF8, FIG4, FIGLA, FOXD3, FOXE1, FOXL1, FREM1, FSHR, FTL, FUS, FUZ, FXYD2, FZD2, GARS1, GATA5, GBA1, GBF1, GCK, GCM2, GCNT2, GDAP1, GDF1, GDF15, GDF2, GDF3, GFI1, GFI1B, GH1, GHRL, GHSR, GJA8, GJB2, GJB3, GJB6, GLRA1, GLS, GLUD1, GNAL, GNAT1, GNB4, GNE, GP1BA, GPD1L, GPD2, GPR161, GPRC5B, GREM2, GRIN2D, GSDME, GSN, GUCA1A, GUCA1B, GUCY2D, H1-4, H4C3, H4C5, H4C9, HABP2, HAL, HARS1, HBA1, HBA2, HBB, HCK, HCN1, HDC, HEPACAM, HLA- DRB1, HMGA2, HMMR, HNMT, HOXD10, HPD, HRG, HSD11B1, HSF4, HSPB1, HSPB8, HTRA1, HTT, IFIH1, IFITM5, IFNGR1, IL13, IL17RD, IMPDH1, IMPG1, IMPG2, INAVA, INS, INSL3, IRF3, IRS1, IRS2, ITGA2B, ITGB3, ITPR3, JAK2, JPH2, KAT8, KBTBD13, KCNA1, KCNA5, KCNC2, KCNE2, KCNJ11, KCNJ18, KCNJ5, KCNK18, KCNK3, KCNK4, KCNN2, KCNN4, KCNQ2, KCNQ4, KCNT1, KCNT2, KCTD1, KCTD17, KDM1A, KIF21A, KIF22, KIF3B, KLF1, KLHL7, KLK11, KRT1, KRT10, KRT12, KRT13, KRT14, KRT16, KRT17, KRT2, KRT4, KRT6A, KRT6B, KRT6C, KRT74, KRT81, KRT83, KRT86, KRT9, LAMA4, LAMB3, LDB3, LEMD2, LGR4, LHCGR, LIM2, LIPC, LITAF, LMBR1, LMBRD2, LORICRIN, LOXL1, LPL, LPP, LRRK2, LRSAM1, LTBP3, LZTR1, MAB21L2, MAF, MAPT, MARS1, MAT1A, MATN3, MB, MBD4, MBL2, MCOLN1, MEF2A, MEFV, MFAP5, MFN2, MIB1, MINPP1, MIP, MLH3, MME, MMP13, MMP19, MPEG1, MPL, MPO, MST1R, MSTO1, MSX2, MTAP, MTHFR, MTMR14, MTNR1B, MTSS2, MUC1, MVD, MVK, MYH14, MYH2, MYH3, MYH6, MYH7, MYH8, MYL2, MYL3, MYLK, MYLK2, MYO3A, MYO7A, MYOC, MYOT, MYOZ2, NAF1, NARS1, NEFH, NEFL, NEK1, NEK8, NEXN, NHERF1, NLRC4, NLRP1, NLRP12, NLRP3, NOBOX, NOP56, NOS3, NOTCH3, NPPA, NR0B2, NR2E3, NRL, NSMF, NUDT15, OAS1, OPA3, OPLAH, OPN1SW, OPTN, ORAI1, OSMR, OVOL2, OXGR1, P2RX2, P4HA2, PABPN1, PAK1, PALLD, PANX1, PARN, PAX4, PCSK9, PDE11A, PDE3A, PDE6B, PDE6H, PDLIM4, PDX1, PERP, PEX6, PIEZO1, PIK3R2, PLAU, PLCD1, PLEC, PLIN1, PLIN4, PMP2, PMVK, PNKD, PNPT1, POGLUT1, POLD1, POLE, POLG, POLG2, POLR3B, POLRMT, POMC, POMP, POT1, POU1F1, PPM1D, PPP1R17, PPP1R3A, PPP2R3C, PRDM6, PRDX3, PRIMPOL, PRKAR1B, PRKCSH, PRKD1, PRNP, PROC, PROK2, PROKR2, PROM1, PRPH,PRPH2, PRSS1, PRX, PSEN2, PSMB10, PSMB9, PSTPIP1, PTDSS1, PTH, PTPN22, PTPRQ, RAB32, RAD54L, RAF1, RBM12, RDH12, RDH5, REEP1, REN, RETN, RHAG, RHO, RHOBTB2, RIGI, RILPL1, RLBP1, RNASEL, RNF213, RNF43, ROBO1, ROBO4, ROM1, RP1, RP1L1, RPA1, RPE65, RRAS2, RTEL1, RTN2, RYR1, RYR2, SAG, SAMD9L, SCN10A, SCN11A, SCN2B, SCN4A, SCN4B, SCN8A, SCN9A, SCNN1B, SCO2, SDHA, SEC23A, SEMA6B, SEPTIN12, SERPINA6, SERPIND1, SERPINE1, SFTPA1, SFTPA2, SFTPC, SH3BP2, SH3TC2, SKI, SLC12A2, SLC12A6, SLC17A3, SLC17A8, SLC17A9, SLC20A2, SLC25A24, SLC26A8, SLC2A2, SLC2A9, SLC30A2, SLC30A8, SLC34A1, SLC36A2, SLC37A4, SLC39A5, SLC3A1, SLC52A1, SLC5A2, SLC6A5, SLC7A9, SLCO2A1, SLFN14, SMAD6, SMAD9, SNTA1, SOD1, SOHLH1, SOST, SP6, SPG7, SPTLC1, SPTSSA, SQSTM1, SREBF1, SSBP1, STING1, STOX1, STUB1, STX16, SYCP3, TARDBP, TBC1D24, TBX2, TBX6, TBXA2R, TBXT, TCAP, TCF3, TCF4 , TECTA, TGFB1, TGFBI, TGM6, THBD, THPO, THSD4, TIE1, TINF2, TLR2, TLR3, TM4SF20, TMC1, TMEM106B, TMEM151A, TMEM163, TMEM43, TMEM63A, TMEM98, TNC, TNFRSF11A, TNFRSF13B, TNNC1, TNNC2, TNNI2, TNNI3, TNNI3K, TNNT1, TNNT2, TNNT3, TNPO3, TNXB, TOR1A, TPM1, TPM2, TPM4, TRAF7, TREX1, TRPC6, TRPM4, TRPM7, TRPV3, TRPV4, TSHR, TTR, TUBA3D, TUBA4A, TUBA8, TUBB1, TUBB3, TWNK, TYR, UBIAD1, UCP3, UFSP2, UGT1A1, UMOD, UNC119, UQCRC1, USP25, VAMP1, VPS16, VSX1, VWF, WASHC5, WDR11, WFS1, WNK4, WNT1, WNT10A, WNT10B, WNT4, XRCC3, YARS1, ZBTB7A, ZFP57, and ZP3.

[0098] Suitably the pathogenic allele may be selected from a gene which causes a cardiac disorder, a skeletal dysplasia or bone disorder, a neurological or neuromuscular disorder, a hematological disorder, an endocrine or metabolic disorder, a renal disorder, or an immunological disorder. Suitably the pathogenic allele may be selected from a gene which causes a dominant negative or gain-of function genetic disorder which may be a cardiac disorder, a neurological or neuromuscular disorder, a hematological disorder, an endocrine or metabolic disorder, a renal disorder, or an immunological disorder.

[0099] Suitably a pathogenic allele which causes a cardiac disorder may be selected from any one of the following genes: ACTC1, ACTN2, AKAP9, ANKRD1, CACNA1C, CALM1, CALM2, CALM3, CAV3, CHRM2, COL1A2, CRYAB, CSRP3, DES, DSC2, EYA4, GATA4, GATAD1, JUP, KCNJ2, KCNJ5, LDB3, LRRC10, MURC / CAVIN4, MYH7, MYL2, MYL3, MYLK2, MYOT, MYOZ2, MYPN, NEBL, NEXN, NKX2-5, PDLIM3, PLN, PRDM16, PRKAG2, PSEN1, PSEN2,RAF1, RBM20, RYR2, SCN4B, SNTA1, TBX20, TCAP, TMEM43, TNNC1, TNNI3, TNNT2, TOR1AIP1, TPM1, TRDN, TTR, TXNRD2, and VCL.

[0100] Suitably, a pathogenic allele which causes a skeletal dysplasia or a bone disorder may be selected from any one of the following genes: ALK2 / ACVR1, CDC73, COL1A1, COL1A2, COL2A1, COMP, COMP, FGF23, FGF23, FGFR3, FGFR3, GCM2, MATN3, and MEN1.

[0101] Suitably, a pathogenic allele which causes a neurological or neuromuscular disorder may be selected from any one of the following genes: AARS1, ADCY5, AFG3L2, ATL1, ATL3, ATN1, ATXN1, ATXN10, ATXN11, ATXN7, ATXN8, ATXN8OS, BEAN1, CACNA1A, CACNA1G, CCDC88C, CHCHD10, CHKB, CHMP2B, COL6A1, COL6A2, COL6A3, C9ORF72, DAB1, DCTN1, DLK1, DNM2, DNM2, ELOVL4, FAT2, FGF14, FTL, FUS, GABRG2, GARS1, GFAP, GNE, GRIN2B, GRN, GRM1, HCN1, HNRNPA1, HNRNPA1, HNRNPA2B1, HNRNPA2B1, HTT, HSPB1, HSPB8, HSPB8, ITPR1, JPH3, KCNA1, KCNC3, KCND3, KCNQ2, KCNQ2, KCNQ2, KCNQ3, KCNQ3, KISS1, KISS1R, KMT2B, MLC1, MKRN3, MFN2, MME, MORC2, MYHC2A, NKX2-1, NOTCH3, NOP56, PABPN1, PDE2A, PHIP, PLD3, PNKD, PPP2R2B, PRKCG, PRNP, PRNP, PRNP, PRRT2, RYR1, SAMD9L, SCN1A, SCN2A, SCN8A, SCN9A, SCN10A, SCN11A, SLC2A1, SOD1, SPTBN2, SPTLC1, SPTLC2, SQSTM1, STUB1, STX1B, TARDBP, TBP, TGM6, TRPC3, TRPV4, TTBK2, and VCP.

[0102] Suitably, a pathogenic allele which causes a hematological disorder may be selected from any one of the following genes: ALAS2, ELANE, EPAS1, GFI1, GP1BA, GP1BA, HAX1, HBA1, HBA2, HBB, MPL, PLAU, SLC25A38, SLFN14 and THPO.

[0103] Suitably, a pathogenic allele which causes an Endocrine and Metabolic Disorders: ABCB6, CACNA1H, CLCN2, CYP19A1, FGF23, FTL, KCNJ5, PLIN1, PPARG, PRSS1, SLC40A1, TSHR, TSHR, and TTR.

[0104] Suitably, a pathogenic allele which causes an Eye Disorder may be selected from any one of the following genes: BEST1, CAPN5, CFI, COL4A1, COL8A2, COL8A2, CTNNA1, EFEMP1, FSCN2, FZD4, GRHL2, IMPDH1, IMPG1, KLHL7, KRT12, MAPKAPK3, NR2E3, OPA3, OPN1LW, OPN1MW, OTX2, OVOL2, PAX6, PLA2G5, PRPF3, PRPF8, RGS9BP, RHO, RP1L1, RP9, SMCHD1, SNRNP200, TCF4, TGFBI, TOPORS, TREX1, VSX1, ZEB1, and ZNF408.

[0105] Suitably, a pathogenic allele which causes a renal disorder may be selected from any one of the following genes: ANGPT2, BSND, CLCNKB, FLT4, FXYD2, GJC2, KCNJ1, PIEZO1, and SLC12A1.

[0106] Suitably, a pathogenic allele which causes an Immunological Disorder may be selected from any one of the following genes: ADAMTS10, ADAMTS17, AP1S3, LTBP2, NLRP3, NOD2, PTPN22, TSC1, and TSC2.

[0107] Suitably, the pathogenic allele may cause other disorders that could benefit from the allele-specific silencing of the therapeutic nucleic acids described herein. Suitably therefore the pathogenic allele may be any one of the following genes: APOE, ATP2A2, COL4A1, GNAS, INHBE, KRT14, KRT5, NKX2-1, SLC22A1, PAFAH1B1, PMVK, PRNP, PSTPIP1, STX16, and TTR.

[0108] Suitably such alleles are pathogenic because they contain one or more mutations compared to a wild type allele of the same gene which cause disease. Suitably there may be many different pathogenic alleles of a given gene, which each containing a different mutation. Suitably therefore a subject having a dominant negative genetic disorder or a gain-of-function genetic disorder may have one of many different pathogenic alleles of a gene which causes said disease. Suitably different subjects having the same disease may have a different pathogenic allele of the same gene. Suitably for the present invention it does not matter which mutation, and therefore which particular pathogenic allele the subject has, only that the pathogenic allele is associated with the target indel allele.

[0109] Suitably the pathogenic allele which causes the dominant negative or gain-of function genetic disorder may be selected from: TNNI3, MYH7, MYL2, TPM1, ACTC1 and TNNT2. In one embodiment, the pathogenic allele which causes the dominant negative or gain-of function genetic disorder may be selected from: TNNI3, MYH7, and TNNT2.

[0110] In one embodiment the pathogenic allele is selected from TNNI3, MYH7, and TNNT2 wherein said allele causes DCM – Dilated cardiomyopathy; HCM – Hypertrophic cardiomyopathy; LVNC – Left ventricular non-compaction cardiomyopathy; and / or RCM – Restrictive cardiomyopathy.

[0111] In one embodiment, the pathogenic allele is TNNI3, wherein said allele causes DCM, RCM, or HCM, suitably HCM, RCM or DCM. Suitably therefore, in such an embodiment, the therapeutic nucleic acid is capable of binding to an target indel allele associated with a pathogenic TNNI3 allele. Suitably which causes HCM, RCM or DCM. Suitably the TNNI3 geneencodes cardiac troponin I protein, which is a myofilament protein important for regulating contractility of cardiac muscle. TNNI3 is commonly mutated in cardiomyopathies, suitably forming a pathogenic TNNI3 allele. Suitably the pathogenic TNNI3 allele comprises one of a number of different pathogenic mutations, mostly missense mutations such as Arg145Gly and Arg162Trp.

[0112] In one embodiment, the pathogenic allele is MYH7, wherein said allele causes DCM, LVNC, RCM, or HCM, suitably HCM or DCM. Suitably therefore, in such an embodiment, the therapeutic nucleic acid is capable of binding to an indel allele associated with a pathogenic MYH7 allele. Suitably which causes HCM or DCM. Suitably the MYH7 gene encodes beta (β)- myosin heavy chain protein, which is a motor protein fundamental to cardiac muscle contraction. MYH7 is commonly mutated in cardiomyopathies, suitably forming a pathogenic MYH7 allele. Suitably the pathogenic MYH7 allele comprises one of a number of different pathogenic mutations, mostly missense mutations such as Arg403Gln and Arg453Cys.

[0113] In one embodiment, the pathogenic allele is TNNT2, wherein said allele causes DCM, LVNC, RCM, or HCM, suitably HCM or DCM. Suitably therefore, in such an embodiment, the therapeutic nucleic acid is capable of binding to an indel allele associated with a pathogenic TNNT2 allele. Suitably which causes HCM or DCM. Suitably the TNNT2 gene encodes cardiac troponin T protein, which is a myofilament protein important for regulating contractility of cardiac muscle. TNNT2 is commonly mutated in cardiomyopathies, suitably forming a pathogenic TNNT2 allele. Suitably the pathogenic TNNT2 allele comprises one of a number of different pathogenic mutations, mostly missense mutations such as Arg92Trp and DelK210.

[0114] In one embodiment, the target indel allele is associated with a pathogenic TNNI3 allele, and is suitably a 4bp insertion or deletion. Suitably said indel allele is located in intron 1 of the pathogenic TNNI3 allele. In such an embodiment, the intronic indel is rs377373012.

[0115] In one embodiment, the target indel allele is associated with a pathogenic MYH7 allele, and is suitably a 1bp insertion or deletion. Suitably said indel allele is located in intron 29 of the pathogenic MYH7 allele. In such an embodiment, the intronic indel is rs34598192. Alternatively, the target indel allele is associated with a pathogenic MYH7 allele, and is suitably a 1bp insertion or deletion. Suitably said indel allele is located in intron 26 of the pathogenic MYH7 allele. In such an embodiment, the intronic indel is rs45504498.

[0116] In one embodiment, the target indel allele is associated with a pathogenic TNNT2 allele, and is suitably a 5bp insertion or deletion. Suitably said indel allele is located in intron 3of the pathogenic TNNT2 allele. Suitably said indel allele is located in exon 4 of the pathogenic TNNT2 allele. In such an embodiment, the intronic indel is rs45533739.

[0117] In one embodiment, the target indel allele associated with a pathogenic TNNI3 allele is an allele of the intronic indel (site) rs377373012. In one embodiment, the target intronic indel allele associated with a pathogenic TNNI3 allele is an allele of the intronic indel (site) rs377373012. Suitably therefore, in one preferred embodiment, the therapeutic nucleic acid is capable of binding to a target indel allele of the intronic indel (site) rs377373012 associated with a pathogenic TNNI3 allele. Suitably therefore, in one preferred embodiment, the therapeutic nucleic acid is capable of binding to a target indel allele of the intronic indel (site) rs377373012 present within pre-mRNA encoding a pathogenic TNNI3 allele. In one embodiment, the target indel allele at indel rs377373012 is either an insertion of the sequence CTGT or a deletion of the sequence CTGT leaving the nucleotide C, suitably therefore the target indel allele at indel rs377373012 may be either CCTGT or C. In one particular embodiment, the target indel allele at indel rs377373012 is CCTGT. In one particular embodiment, the target indel allele at indel rs377373012 is C. Suitably, rs377373012 is located on chromosome 19 at position 55157354 of human genome GRCh38.p14. Suitably, rs377373012 is located at position 414 of gene TNNI3 (SEQ ID 222). Suitably in such embodiments, the therapeutic nucleic acid is an antisense oligonucleotide and comprises or may comprise a sequence according to SEQ ID NO: 87 (E3) or SEQ ID NO: 101 (E17), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. It will be appreciated that the therapeutic nucleic acid may comprise a sequence which targets and binds to the insertion allele or to the deletion allele at indel site rs377373012. Suitably the therapeutic nucleic acids described herein bind to one of the indel alleles i.e. to the insertion or to the deletion, however therapeutic nucleic acids of the invention may equally target and bind to the alternative indel allele to that bound by the sequences described herein. Suitably therefore the invention extends to a therapeutic nucleic acid comprising a sequence which targets and binds to the alternative indel allele to the indel allele bound by the nucleic acids recited herein, such as SEQ ID NO: 87 (E3) or SEQ ID NO: 101 (E17). Suitably in such embodiments, the therapeutic nucleic acid is an antisense oligonucleotide and comprises or may comprise a sequence according to any of the nucleic acids recited herein, such as SEQ ID NO: 87 (E3) or SEQ ID NO: 101 (E17), or a sequence which targets and binds to the alternative indel allele at indel rs377373012.

[0118] Accordingly, there is provided a therapeutic nucleic acid capable of binding to an allele of the intronic indel rs377373012, suitably to a target indel allele of the intronic indel rs377373012. In one embodiment, the therapeutic nucleic acid comprises a sequence according to SEQ ID NO: 101 (E17). In one embodiment, the therapeutic nucleic acid comprises a sequence according to SEQ ID NO: 87 (E3). Optionally wherein the therapeutic nucleic acid is an antisense oligonucleotide.

[0119] In one embodiment, the target indel associated with a pathogenic MYH7 allele is an allele of the intronic indel (site) rs34598192. In one embodiment, the target intronic indel allele associated with the pathogenic MYH7 allele is an allele of the intronic indel (site) rs34598192. Suitably therefore, in one preferred embodiment, the therapeutic nucleic acid is capable of binding to a target indel allele of the intronic indel (site) rs34598192 associated with a pathogenic MYH7 allele. Suitably therefore, in one preferred embodiment, the therapeutic nucleic acid is capable of binding to a target indel allele of the intronic indel (site) rs34598192 present within pre-mRNA encoding a pathogenic MYH7 allele. In one embodiment the target indel allele at indel rs34598192 is either an insertion of the sequence G or a deletion of the sequence G leaving the nucleotide T, suitably therefore the target indel allele at indel rs34598192 may be either TG or T. In one particular embodiment, the target indel allele at indel rs34598192 is TG. In one particular embodiment, the target indel allele at indel rs34598192 is T. Suitably, rs34598192 is located on chromosome 14 at position 23419114 of human genome GRCh38.p14. Suitably, rs34598192 is located at position 16546 or 16547 of gene MYH7 (SEQ ID 221). Suitably in such embodiments, the therapeutic nucleic acid is an antisense oligonucleotide and comprises or may comprise a sequence according to SEQ ID 66 (D7) or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. It will be appreciated that the therapeutic nucleic acid may comprise a sequence which targets and binds to the insertion allele or to the deletion allele at indel site rs34598192. Suitably the therapeutic nucleic acids described herein bind to one of the indel alleles i.e. to the insertion or to the deletion, however therapeutic nucleic acids of the invention may equally target and bind to the alternative indel allele to that bound by the sequences described herein. Suitably therefore the invention extends to a therapeutic nucleic acid comprising a sequence which targets and binds to the alternative indel allele to the indel allele bound by the nucleic acids recited herein, such as SEQ ID 66 (D7). Suitably in suchembodiments, the therapeutic nucleic acid is an antisense oligonucleotide and comprises or may comprise a sequence according to any of the nucleic acids recited herein, such as SEQ ID 66 (D7), or a sequence which targets and binds to the alternative indel allele at indel rs34598192.

[0120] Accordingly, there is provided a therapeutic nucleic acid capable of binding to an allele of the intronic indel rs34598192, suitably to a target indel allele of the intronic indel rs34598192. In one embodiment, the therapeutic nucleic acid comprises a sequence according to SEQ ID NO: 66 (D7). Optionally wherein the therapeutic nucleic acid is an antisense oligonucleotide.

[0121] In one embodiment, the target indel associated with a pathogenic MYH7 allele is an allele of the intronic indel (site) rs45504498. In one embodiment, the target intronic indel allele associated with the pathogenic MYH7 allele is an allele of the intronic indel (site) rs45504498. Suitably therefore, in one preferred embodiment, the therapeutic nucleic acid is capable of binding to a target indel allele of the intronic indel (site) rs45504498 associated with a pathogenic MYH7 allele. Suitably therefore, in one preferred embodiment, the therapeutic nucleic acid is capable of binding to a target indel allele of the intronic indel (site) rs45504498 present within pre-mRNA encoding a pathogenic MYH7 allele. In one embodiment the target indel allele at indel rs45504498 is either an insertion of the sequence G or a deletion of the sequence G leaving the nucleotide T, suitably therefore the target indel allele at indel rs45504498 may be either TG or T. In one particular embodiment, the target indel allele at indel rs45504498 is TG. In one particular embodiment, the target indel allele at indel rs45504498 is T. Suitably, rs45504498 is located on chromosome 14 at position 23420236 of human genome GRCh38.p14. Suitably, rs45504498 is located at position 15423 of gene MYH7 (SEQ ID 221). It will be appreciated that the therapeutic nucleic acid may comprise a sequence which targets and binds to the insertion allele or to the deletion allele at indel site rs45504498. Suitably the therapeutic nucleic acids described herein bind to one of the indel alleles i.e. to the insertion or to the deletion, however therapeutic nucleic acids of the invention may equally target and bind to the alternative indel allele to that bound by the sequences described herein.

[0122] Accordingly, there is provided a therapeutic nucleic acid capable of binding to an allele of the intronic indel rs45504498, suitably to a target indel allele of the intronic indel rs45504498. Optionally wherein the therapeutic nucleic acid is an antisense oligonucleotide.

[0123] In one embodiment, the target indel allele associated with the pathogenic TNNT2 allele is an allele of the intronic indel (site) rs45533739. In one embodiment, the target intronic indelallele associated with a pathogenic TNNT2 allele is an allele of the intronic indel (site) rs45533739. Suitably therefore, in one preferred embodiment, the therapeutic nucleic acid is capable of binding to a target indel allele of the intronic indel (site) rs45533739 present within a pathogenic TNNT2 allele. Suitably therefore, in one preferred embodiment, the therapeutic nucleic acid is capable of binding to a target indel allele of the intronic indel (site) rs45533739 present within pre-mRNA encoding a pathogenic TNNT2 allele. In one embodiment the target indel allele at indel rs45533739 is either an insertion of the sequence AGAAG or a deletion of the sequence AGAAG leaving the nucleotide C, suitably therefore the target indel allele at indel rs45533739 may be either CAGAAG or C. In one particular embodiment, the target indel allele at indel rs45533739 is C. In one particular embodiment, the target indel allele at indel rs45533739 is CAGAAG. Suitably, rs45533739 is located on chromosome 1 at position 201372047 of human genome GRCh38.p14. Suitably, rs45533739 is located at position 5708 of gene TNNT2 (SEQ ID NO: 220). Suitably in such embodiments, the therapeutic nucleic acid is an antisense oligonucleotide and comprises or may comprise a sequence according to SEQ ID NO: 156 (F12), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. It will be appreciated that the therapeutic nucleic acid may comprise a sequence which targets and binds to the insertion allele or to the deletion allele at indel site rs45533739. Suitably the therapeutic nucleic acids described herein bind to one of the indel alleles i.e. to the insertion or to the deletion, however therapeutic nucleic acids of the invention may equally target and bind to the alternative indel allele to that bound by the sequences described herein. Suitably therefore the invention extends to a therapeutic nucleic acid comprising a sequence which targets and binds to the alternative indel allele to the indel allele bound by the nucleic acids recited herein, such as SEQ ID NO: 156 (F12). Suitably in such embodiments, the therapeutic nucleic acid is an antisense oligonucleotide and comprises or may comprise a sequence according to any of the nucleic acids recited herein, such as SEQ ID NO: 156 (F12), or a sequence which targets and binds to the alternative indel allele at indel rs45533739.

[0124] Accordingly, there is provided a therapeutic nucleic acid capable of binding to an allele of the intronic indel rs45533739, suitably to a target indel allele of the intronic indel rs45533739. In one embodiment, the therapeutic nucleic acid comprises a sequence accordingto SEQ ID NO: 156 (F12). Optionally wherein therapeutic nucleic acid is an antisense oligonucleotide.

[0125] Dominant Negative Genetic Disorder and Gain-of-Function Genetic Disorder

[0126] A dominant genetic disorder is one in which a single copy (allele) of a mutated gene is enough to cause disease. A dominant negative genetic disorder means that the mutated allele, when expressed, produces mutated protein encoded by the mutated allele which negatively impacts the function of the wild type protein and causes pathology. A gain-of-function genetic disorder means that the mutated allele, when expressed, produces mutated protein that has a different activity or an increased activity compared to the wild type protein which causes pathology.

[0127] Suitably the genetic disorder is dominant. Suitably the gain-of-function disorder is a dominant gain-of-function disorder. Suitably the genetic disorder is inherited. Suitably the genetic disorder is an autosomal dominant genetic disorder.

[0128] Suitably the genetic disorder is caused by a gain-of-function or by a dominant negative mutation. Suitably the genetic disorder may be a Skeletal Dysplasia or Bone Disorder, suitably selected from any one of the following disorders: Achondrogenesis type II, Achondroplasia, Fibrodysplasia ossificans progresssiva, Hypochondroplasia, Multiple epiphyseal dysplasia type, Osteogenesis imperfecta, Pseudoachondroplasia, Autosomal dominant hypophosphatemic rickets, and Autosomal dominant primary hyperparathyroidism.

[0129] Suitably the genetic disorder may be a Neurological or Neuromuscular Disorder, suitably selected from any one of the following disorders: Alexander disease, Amyotrophic lateral sclerosis, Autosomal dominant centronuclear myopathy, Autosomal dominant Charcot- Marie-Tooth disease type 2, Autosomal dominant congenital benign spinal muscular atrophy, Autosomal dominant distal axonal motor neuropathy-myofibrillar myopathy syndrome, Benign familial infantile epilepsy, Benign familial neonatal epilepsy, Benign familial neonatal-infantile seizures, Benign hereditary chorea, Benign paroxysmal torticollis of infancy, Benign Samaritan congenital myopathy, Cerebral autosomal dominant arteriopathy-subcortical infarcts- leukoencephalopathy, Congenital muscular dystrophy type Ullrich, Frontotemporal dementia with motor neuron disease, Generalized epilepsy with febrile seizures-plus, GRN Frontotemporal dementia, Hereditary sensory and autonomic neuropathy type, Hereditary sodium channelopathy-related small fibers neuropathy, Hereditary motor and sensory neuropathy type, Inclusion body myopathy with Paget disease of bone and frontotemporaldementia, Infantile convulsions and choreoathetosis, Megaconial congenital muscular dystrophy, Megalencephalic leukoencephalopathy with subcortical cysts, Paroxysmal dystonic choreathetosis with episodic ataxia and spasticity, Paroxysmal extreme pain disorder, Paroxysmal kinesigenic dyskinesia, Paroxysmal non-kinesigenic dyskinesia, Perry syndrome, Spinocerebellar ataxia type 1, Spinocerebellar ataxia type 10, Spinocerebellar ataxia type 11, Spinocerebellar ataxia type 12, Spinocerebellar ataxia type 14, Spinocerebellar ataxia type 13, Spinocerebellar ataxia type 6, Spinocerebellar ataxia type 15, Spinocerebellar ataxia type 17, Spinocerebellar ataxia type 19, Spinocerebellar ataxia type 27, Spinocerebellar ataxia type 28, Spinocerebellar ataxia type 31, Spinocerebellar ataxia type 34, Spinocerebellar ataxia type 35, Spinocerebellar ataxia type 36, Spinocerebellar ataxia type 37, Spinocerebellar ataxia type 38, Spinocerebellar ataxia type 41, Spinocerebellar ataxia type 42, Spinocerebellar ataxia type 43, Spinocerebellar ataxia type 44, Spinocerebellar ataxia type 45, Spinocerebellar ataxia type 46, Spinocerebellar ataxia type 47, Spinocerebellar ataxia type 48, Spinocerebellar ataxia type 50, Spinocerebellar ataxia type 7, Spinocerebellar ataxia type 8, Spinocerebellar ataxia type 8, Huntington's disease, Huntington disease-like, Dentatorubral pallidoluysian atrophy, Dystonia 28, Familial Creutzfeldt–Jakob disease, Gerstmann–Sträussler–Scheinker syndrome, Fatal familial insomnia, Genetic central precocious puberty in female, GRIN2B-related neurodevelopmental disorder, Hereditary inclusion body myopathy, Neuroferritinopathy, Oculopharyngeal muscular dystrophy, PHIP-related behavioral problems-intellectual disability- obesity-dysmorphic features syndrome, and Pigmented paravenous retinochoroidal atrophy.

[0130] Suitably the genetic disorder may be a Cardiac Disorder, suitably selected from any one of the following disorders: Andersen-Tawil syndrome, Cardiac-valvular Ehlers-Danlos syndrome, Catecholaminergic polymorphic ventricular tachycardia, Dilated Cardiomyopathy (DCM), Familial isolated restrictive cardiomyopathy, Familial progressive cardiac conduction defect, Hypertrophic Cardiomyopathy (HCM), Left ventricular noncompaction, Noonan syndrome, Restrictive Cardiomyopathy (RCM), Romano-Ward syndrome, and Short QT syndrome.

[0131] Suitably the genetic disorder may be a hematological Disorder, suitably selected from any one of the following disorders: Autosomal dominant macrothrombocytopenia, Autosomal dominant secondary polycythemia, Autosomal dominant thrombocytopenia with platelet secretion defect, Familial thrombocytosis, Pseudo-von Willebrand disease, Quebec platelet disorder, Sideroblastic anemia, and Severe congenital neutropenia.

[0132] Suitably the genetic disorder may be an Endocrine or Metabolic Disorder, suitably selected from any one of the following disorders: Aromatase excess syndrome, Autosomal dominant hypophosphatemic rickets, Euthyroid dysprealbuminemic hyperthyroxinemia, Familial gestational hyperthyroidism, Familial hyperaldosteronism type, Familial hyperthyroidism due to mutations in TSH receptor, Familial pseudohyperkalemia, Hereditary hyperferritinemia-cataract syndrome, Hereditary hemochromatosis type 4, Hereditary pancreatitis, PPARG-related familial partial lipodystrophy, and PLIN1-related familial partial lipodystrophy.

[0133] Suitably the genetic disorder may be an Eye Disorder, suitably selected from any one of the following disorders: Autosomal dominant keratitis, Autosomal dominant neovascular inflammatory vitreoretinopathy, Autosomal dominant optic atrophy and cataract, Benign concentric annular macular dystrophy, Best vitelliform macular dystrophy, Blue cone monochromatism, Bradyopsia, Butterfly-shaped pigment dystrophy, Facioscapulohumeral dystrophy, Familial benign flecked retina, Familial drusen, Familial exudative vitreoretinopathy, Fuchs endothelial corneal dystrophy, Granular corneal dystrophy type, Meesman corneal dystrophy, Occult macular dystrophy, Posterior polymorphous corneal dystrophy, Retinal arterial tortuosity, Retinal vasculopathy with cerebral leukoencephalopathy and systemic manifestations, Retinitis pigmentosa, Martinique crinkled retinal pigment epitheliopathy.

[0134] Suitably the genetic disorder may be a Renal Disorder, suitably selected from any one of the following disorders: Autosomal dominant primary hypomagnesemia with hypocalciuria, Bartter syndrome, and Milroy disease.

[0135] Suitably the genetic disorder may be an Immunological Disorder, suitably selected from any one of the following disorders: Rheumatoid arthritis, Blau syndrome, Generalized pustular psoriasis, Muckle-Wells syndrome, Tuberous sclerosis, and Weill-Marchesani syndrome.

[0136] Suitably the genetic disorder may also be selected from any of the following disorders: ATTRV122I amyloidosis, ATTRV30M amyloidosis, Brain-lung-thyroid syndrome, Darier disease, Lissencephaly due to LIS1 mutation, Localized epidermolysis bullosa simplex, PAPA syndrome, Pontine autosomal dominant microangiopathy with leukoencephalopathy, Porokeratosis of Mibelli, PrP systemic amyloidosis, Lipoprotein glomerulopathy, Pseudohypoparathyroidism and Obesity.

[0137] Suitably the present invention treats subjects who are heterozygous for a pathogenic allele, suitably therefore who have one copy of a wild type allele and one copy of a pathogenic allele, suitably which causes a dominant negative disorder or a gain-of-function disorder. Suitably the present invention treats dominant genetic disorders in which suppression of the wild type allele is harmful to the subject. Suitably in which the suppression of the wild type allele is not tolerated by the subject. Suitably in which the suppression of the wild type allele causes a worsening of the disorder in the subject. Suitably therefore any therapeutic must be allele-specific. Suitably the present invention treats subjects having a dominant negative pathogenic allele, or a gain-of-function pathogenic allele. Suitably the present invention treats subjects having a dominant negative pathogenic allele, or a gain-of-function pathogenic allele in a gene that is tolerant of single copy loss-of-function (i.e. a gene that is haplosufficient). Suitably by haplosufficient it is meant that one copy of a wild type allele in the subject has enough function to produce a wild type phenotype in the subject. Suitably therefore, when treated with the therapeutic nucleic acids of the present invention, expression of the pathogenic allele is suppressed or silenced, and expression of the wild type allele is sufficient to restore the wild type phenotype.

[0138] Suitably the disorder is a cardiomyopathy, suitably a dominant negative cardiomyopathy, suitably it is an inherited cardiomyopathy, suitably an autosomal dominant cardiomyopathy, suitably an autosomal dominant negative cardiomyopathy.

[0139] Suitably the cardiomyopathy is selected from: hypertrophic cardiomyopathy, dilated cardiomyopathy, arrythmogenic right ventricular cardiomyopathy, restrictive cardiomyopathy, left ventricular noncompaction cardiomyopathy, and peripartum cardiomyopathy. In one embodiment the cardiomyopathy is selected from: hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), and restrictive cardiomyopathy (RCM). In one embodiment, the cardiomyopathy is HCM, DCM or RCM. In one embodiment, there is provided a therapeutic nucleic acid capable of binding to a target indel allele associated with a pathogenic allele, wherein the pathogenic allele causes a dominant negative genetic disorder or gain-of function genetic disorder, and wherein the disorder is a cardiomyopathy. In one embodiment, there is provided a therapeutic nucleic acid capable of binding to a target indel allele associated with a pathogenic allele, wherein the pathogenic allele causes a dominant negative cardiomyopathy. In one embodiment, there is provided a therapeutic nucleic acid capable of binding to a target indelallele associated with a pathogenic allele, wherein the pathogenic allele causes HCM, DCM or RCM.

[0140] Suitably, the disorder according to the first to eighth aspects of the invention is not Huntington’s Disease.

[0141] Huntington’s Disease

[0142] Huntington disease (HD) is an autosomal dominant negative disorder caused by the expansion of the glutamine encoding CAG tract in exon one of the huntingtin HTT gene forming a pathogenic HTT allele, which leads to the production of polyglutamine expanded mutant huntingtin (mtHTT) protein. HD manifests in people with 36 or more CAG repeats with full penetrance seen with 40 or more. Suitably therefore the pathogenic HTT allele comprises 36 or more CAG repeats. Suitably said pathogenic HTT allele causes Huntington disease (HD).

[0143] The age of disease onset is influenced inversely by the length of the expansion, and symptoms do not typically develop until midlife. Symptoms of HD, including psychiatric disturbance, cognitive dysfunction, and the progressive loss of voluntary motor control, worsen over time until death, typically 15–20 years after motor onset. There is no cure for HD and no disease-modifying treatments available. There is strong evidence that reduction of mtHTT protein could provide such a treatment. Conditional inactivation of mtHTT in adult, symptomatic HD mice results in the recovery of motor function and clearance of toxic protein aggregates, therefore ASO therapies for HD which lower mutant HTT levels have been developed. However these ASOs have been designed to non-selectively target both wild type and pathogenic HTT transcripts, or to target CAG expansion-associated SNPs in the pathogenic HTT transcript, and they have not yet been successful in human trials. The present invention also sets out to provide an alternative HD treatment which addresses these issues.

[0144] In alternative aspects of the present invention, there is provided a therapeutic nucleic acid which is capable of binding to an intronic indel allele associated with a pathogenic HTT allele. In accordance with alternative aspects of the invention, the therapeutic nucleic acid is capable of binding to an intronic indel allele present in a pathogenic HTT allele which causes Huntington’s disease. Suitably the intronic indel allele is an allele of an intronic indel selected from: rs59464879 (G / GGAGTT), rs751205475 (G / GTTGTATGGTTTGGAGGTGCTC, SEQ ID NO: 233) and rs78373442 (G / GAATATT).

[0145] Suitably the therapeutic nucleic acid is capable of binding to a target indel allele of an intronic indel (site) selected from rs59464879, rs751205475 and rs78373442 present in a pathogenic HTT allele which causes Huntington’s disease. Suitably the therapeutic nucleic acid is capable of binding to a target intronic indel allele present in pre-mRNA encoding the pathogenic HTT allele which causes Huntington’s disease. Suitably the therapeutic nucleic acid is capable of binding to a target indel allele of an intronic indel (site)selected from rs59464879, rs751205475 and rs78373442 present in pre-mRNA encoding the pathogenic HTT allele which causes Huntington’s disease.

[0146] In one embodiment the target indel allele at indel rs59464879 is either an insertion of the sequence GAGTT or a deletion of the sequence GAGTT leaving the nucleotide G, suitably therefore the target indel allele at indel rs59464879 is either GGAGTT or G. In one particular embodiment, the target indel allele at indel rs59464879 is G. In one particular embodiment, the target indel allele at indel rs59464879 is GGAGTT. Suitably, rs59464879 is located on chromosome 4 at position 3239417 of human genome GRCh38.p14. Suitably, rs59464879 is located at position 198056 of gene HTT (SEQ ID NO: 223). In one embodiment the target indel allele at indel rs751205475 is either an insertion of the sequence TTGTATGGTTTGGAGGTGCTC (SEQ ID NO: 234) or a deletion of the sequence TTGTATGGTTTGGAGGTGCTC (SEQ ID NO: 235) leaving the nucleotide G, suitably therefore the target indel allele at indel rs751205475 is either GTTGTATGGTTTGGAGGTGCTC (SEQ ID NO: 236) or G. In one particular embodiment, the target indel allele at indel rs751205475 is G. In one particular embodiment, the target indel allele at indel rs751205475 is GTTGTATGGTTTGGAGGTGCTC. Suitably, rs751205475 is located on chromosome 4 at position 3099609 of human genome GRCh38.p14. Suitably, rs751205475 is located at position 58248 of gene HTT (SEQ ID NO: 223). In one embodiment the target indel allele at indel rs78373442 is either an insertion of the sequence AATATT, or a deletion of the sequence AATATT leaving the nucleotide G, suitably therefore the target indel allele at indel rs78373442 is either GAATATT or G. In one particular embodiment, the target indel allele at indel rs78373442 is GAATATT. In one particular embodiment, the target indel allele at indel rs78373442 is G. Suitably, rs78373442 is located on chromosome 4 at position 3064874 of human genome GRCh38.p14. Suitably, rs78373442 is located at position 23512 of gene HTT (SEQ ID NO: 223). Suitably the therapeutic nucleic acid is capable of binding to either of the alleles located at each intronic indel site.

[0147] Suitably the therapeutic nucleic acid comprises a sequence according to SEQ ID NO: 188-219, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably the therapeutic nucleic acid is an ASO and comprises a sequence selected from: SEQ ID NO: 188-191, 199-202 and 212-215, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably the therapeutic nucleic acid comprises selected from SEQ ID NO: 188, 199 and 212, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Suitably the therapeutic nucleic acid consists of a sequence selected from SEQ ID NO: 188, 199 and 212.

[0148] In one embodiment, the therapeutic nucleic acid comprises a sequence selected from any one of SEQ ID NO: 188-198, which target and bind to a target indel allele of the intronic indel (site) rs59464879. Suitably which is associated with the pathogenic allele HTT.

[0149] In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 188-191, which target and bind to a target indel allele of the intronic indel (site) rs59464879. Suitably which is associated with the pathogenic allele HTT.

[0150] In one embodiment, the therapeutic nucleic acid comprises a sequence selected from any one of SEQ ID NO: 199-211, which target and bind to a target indel allele of the intronic indel (site) rs751205475. Suitably which is associated with the pathogenic allele HTT.

[0151] In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 199-202, which target and bind to a target indel allele of the intronic indel (site) rs751205475. Suitably which is associated with the pathogenic allele HTT.

[0152] In one embodiment, the therapeutic nucleic acid comprises a sequence selected from any one of SEQ ID NO: 212-219, which target and bind to a target indel allele of the intronic indel (site) rs78373442. Suitably which is associated with the pathogenic allele HTT.

[0153] In one embodiment, the therapeutic nucleic acid is an ASO and comprises a sequence selected from any one of SEQ ID NO: 212-215, which target and bind to a target indel allele of the intronic indel (site) rs78373442. Suitably which is associated with the pathogenic allele HTT.

[0154] Suitably the therapeutic nucleic acid is efficacious in reducing the level, expression and / or activity of a pathogenic HTT allele (or a protein product thereof), and is capable of mediating allele-specific knockdown of the pathogenic HTT allele.

[0155] In one embodiment, there is provided a therapeutic nucleic acid comprising a sequence according to SEQ ID 188, which is capable of binding to a target indel allele of the intronic indel rs59464879 associated with a pathogenic HTT allele. In one embodiment, there is provided a therapeutic nucleic acid comprising a sequence according to SEQ ID 199, which is capable of binding to a target indel allele of the intronic indel rs751205475 associated with a pathogenic HTT allele. In one embodiment, there is provided a therapeutic nucleic acid comprising a sequence according to SEQ ID 212, which is capable of binding to a target indel allele of the intronic indel rs78373442 associated with a pathogenic HTT allele. It will be appreciated that the therapeutic nucleic acid may comprise a sequence which targets and binds to the insertion allele or to the deletion allele at indel sites rs59464879, rs751205475, and rs78373442. Suitably the therapeutic nucleic acids described herein bind to one of the indel alleles i.e. to the insertion or to the deletion, however therapeutic nucleic acids of the invention may equally target and bind to the alternative indel allele to that bound by the sequences described herein. Suitably therefore the invention extends to a therapeutic nucleic acid comprising a sequence which targets and binds to the alternative indel allele to the indel allele bound by the nucleic acids recited herein, such as SEQ ID NO: 188, 199 or 212. Suitably in such embodiments, the therapeutic nucleic acid is an antisense oligonucleotide and comprises a sequence according to any of the nucleic acids recited herein, such as SEQ ID NO: 188, 199, 212, or a sequence which targets and binds to the alternative indel allele at indel rs59464879, rs751205475, and rs78373442 respectively.

[0156] Advantageously the inventors have determined and identified novel intronic indel alleles which associate with pathogenic HTT alleles, and designed and tested ASOs which target and bind to these intronic indel alleles in HTT pre-mRNA with high specificity. The ASO provides an alternate allele-specific therapy for treating Huntington’s disease.

[0157] Suitably any other features or aspects described herein in relation to the first to eighth aspects apply equally to the aspects relating to treatment of Huntington’s disease.

[0158] Related products

[0159] The invention also provides related products comprising or encoding the agents and in particular the therapeutic nucleic acids according to the invention. Accordingly, the inventionprovides a conjugate comprising the therapeutic nucleic acid covalently linked to a delivery group, or a delivery particle comprising the therapeutic nucleic acid.

[0160] Conjugate

[0161] Suitably a conjugate of the invention may comprise any delivery group capable of aiding delivery or uptake of the therapeutic nucleic acid into a target cell. Suitable such delivery groups may include: antibodies, peptides, small molecule chemicals, polymers, lipids, or the like.

[0162] Suitably the delivery group is a peptide. Suitably, the peptide may be selected from viral proteins such as VP22 (derived from herpes virus tegument protein), snake venom protein such as CyLOP-1 (derived from crotamin), cell adhesion glycoproteins such as pVEC (derived from murine vascular endothelial-cadherin protein), Penetratin (Antennapedia homeodomain), Tat (human immunodeficiency virus transactivating regulatory protein) or reverse Tat, for example. In some embodiments, the therapeutic nucleic acid is conjugated to a cell penetrating peptide (CPP) to enhance therapeutic uptake.

[0163] Suitable CPPs are known in the art. The use of arginine-rich peptide carriers is particularly useful. Certain arginine-based peptide carriers have been shown to be highly effective at delivery of antisense compounds into primary cells including muscle cells (Marshall, Oda et al. 2007; Jearawiriyapaisarn, Moulton et al. 2008; Wu, Moulton et al. 2008). Furthermore, compared to other peptides, the arginine peptide carriers when conjugated to an antisense oligonucleotide, demonstrate an enhanced ability to alter transcript levels (Marshall, Oda et al. 2007). Suitably, the cell penetrating peptide may be selected from those peptides described in WO2015075747, WO2013030569, WO2009147368, US20120289457, or US20160237426, for example.

[0164] By conjugated it is meant that the therapeutic nucleic acid is covalently linked to the delivery group. Suitably, conjugation of the delivery group to the therapeutic nucleic acid may be at any position suitable for forming a covalent bond between the delivery group and the therapeutic nucleic acids or between a linker and the therapeutic nucleic acid. For example, conjugation of a delivery group may be at the 3' end of the therapeutic nucleic acid. Alternatively, conjugation of a delivery group to the therapeutic nucleic acid may be at the 5' end of the therapeutic nucleic acid. Alternatively, a delivery group may be conjugated to the therapeutic nucleic acid through any of the intersubunit linkages.

[0165] Suitably, the delivery group is covalently coupled at its N-terminal or C-terminal residue to the 3' or 5' end of the therapeutic nucleic acid. Suitably, the delivery group is coupled at its C-terminal residue to the 5' end of the therapeutic nucleic acid.

[0166] Optionally, where the therapeutic nucleic acid comprises phosphorus-containing intersubunit linkages, and the delivery group is a peptide, the peptide may be conjugated to the therapeutic nucleic acid via a covalent bond to the phosphorous of the terminal linkage group.

[0167] Alternatively, when the delivery group is a peptide, and the therapeutic nucleic acid is a morpholino, the peptide may be conjugated to the nitrogen atom of the 3' terminal morpholino group of the oligomer.

[0168] Optionally, the delivery group may be conjugated to the therapeutic nucleic acid via a linker. Optionally, the linker may comprise one or more of: an optionally substituted piperazinyl moiety, a beta alanine, glycine, proline, 6-aminohexanoic acid residue, lysine and / or arginine in any combination.

[0169] Alternatively, the delivery group may be conjugated directly to the therapeutic nucleic acid without a linker.

[0170] Suitably the conjugate may further comprise a homing moiety. Suitably, the homing moiety is selective for a selected mammalian tissue, i.e., the same tissue being targeted by the therapeutic nucleic acid. Suitably, the homing moiety is selective for muscle tissue, suitably cardiac tissue.

[0171] Suitably, the homing moiety is a homing peptide. Suitable homing peptides are disclosed in 'Effective Dystrophin Restoration by a Novel Muscle-Homing Peptide-Morpholino Conjugate in Dystrophin-Deficient mdx Mice' Gao et al. Mol Ther. 2014 Jul; 22(7): 1333-1341, for example.

[0172] Suitably, the delivery peptide and the homing peptide may be formed as a chimeric fusion protein. Suitably, the conjugate may comprise a chimeric peptide formed from a cell penetrating peptide and a cardiac-specific homing peptide. Optionally, the conjugate may be of the form: delivery peptide-homing peptide-therapeutic nucleic acid or of the form: homing peptide-carrier peptide-therapeutic nucleic acid. Optionally comprising linkers located between each element.

[0173] Suitably, the therapeutic nucleic acid may be conjugated to a delivery group that enhances the solubility of the therapeutic nucleic acid. Suitably the solubility in an aqueousmedium. Suitably, a delivery group that enhances solubility may be conjugated to the therapeutic nucleic acid in addition to a delivery group operable to transport the therapeutic nucleic acid.

[0174] Suitably, the delivery group that enhances solubility and the delivery group that transports the therapeutic nucleic acid may be formed as a chimeric fusion protein.

[0175] Suitable delivery groups that enhance the solubility of a therapeutic nucleic acid are polymers, such as polyethylene glycol, or triethylene glycol.

[0176] Particle

[0177] Alternatively or additionally the therapeutic nucleic acid of the invention, or indeed the conjugate thereof, or a vector comprising or encoding said therapeutic nucleic acid, may be comprised within a particle, suitable a delivery particle. Suitably the delivery particle also is capable of aiding delivery or uptake of the therapeutic nucleic acid into a target cell. Suitable delivery particles may include: nanoparticles, liposomes, exosomes or the like.

[0178] For example, a variety of supramolecular nanocarriers can be used to deliver nucleic acids. Example nanocarriers include, but are not limited to liposomes, cationic polymer complexes and various polymeric compounds. Complexation of nucleic acids with various polycations is another approach for intracellular delivery; this includes use of PEGylated polycations, polyethyleneamine (PEI) complexes, cationic block co-polymers, and dendrimers. Several cationic nanocarriers, including PEI and polyamidoamine dendrimers help to release contents from endosomes. Other approaches include use of polymeric nanoparticles, microspheres, liposomes, dendrimers, biodegradable polymers, conjugates, prodrugs, inorganic colloids such as sulfur or iron, antibodies, implants, biodegradable implants, biodegradable microspheres, osmotically controlled implants, lipid nanoparticles, emulsions, oily solutions, aqueous solutions, biodegradable polymers, poly(lactide-coglycolic acid), poly(lactic acid), liquid depot, polymer micelles, quantum dots and lipoplexes. In some embodiments, an oligonucleotide is conjugated to another molecule.

[0179] In some embodiments, the therapeutic nucleic acid, the conjugate thereof, or a vector comprising or encoding said therapeutic nucleic acid, is comprised within a liposome particle.

[0180] Suitably, the delivery group or particle has the capability of inducing cell penetration of the therapeutic nucleic acid within at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of cells of a given cell culture population. Suitably, the carrier has the capability of inducing cell penetration of the therapeutic nucleic acid within at least 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of cardiac cells.

[0181] Vector

[0182] Further aspects of the invention may also include a vector comprising or encoding the therapeutic nucleic acid according to the invention. Suitably, the vector may be an expression vector. The therapeutic nucleic acid is typically subcloned into the vector in an antisense orientation (i.e., RNA transcribed from the inserted therapeutic nucleic acid will be of an antisense orientation to the target indel allele of interest).

[0183] Optionally the therapeutic nucleic acid may be present in the vector in the form of an RNA sequence, suitably in the form of a shRNA sequence. Suitably therefore the therapeutic nucleic acid is expressed from the vector as an shRNA.

[0184] Suitably the vector may comprise an shRNA sequence according to any of SEQ ID NOs: 81-84, 142-145, 184-187, 195-198, 208-211, and 216-219 or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. Optionally the vector may comprise one or more of said shRNA sequences, which may suitably target and bind to different target indel alleles. Optionally the different target indel alleles may be associated with the same pathogenic allele.

[0185] In one embodiment the vector comprises an shRNA sequence encoding a therapeutic nucleic acid which targets and binds to an indel allele of an intronic indel associated with a pathogenic MYH7 allele, wherein the shRNA sequence comprises any of SEQ ID NO:81-84, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. In one embodiment the vector comprises an shRNA sequence encoding a therapeutic nucleic acid which targets and binds to an indel allele of an intronic indel associated with a pathogenic TNNI3 allele, wherein the shRNA sequence comprises any of SEQ ID NO: 142-145, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. In one embodiment the vector comprises an shRNA sequence encoding a therapeutic nucleic acid which targets and binds to an indel allele of an intronic indel associated with a pathogenic TNNT2 allele, wherein the shRNA sequence comprises any of SEQ ID NO: 184-187 or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto. In one embodiment thevector comprises an shRNA sequence encoding a therapeutic nucleic acid which targets and binds to an indel allele of an intronic indel associated with a pathogenic HTT allele, wherein the shRNA sequence comprises any of SEQ ID NO: 195-198, 208-211, and 216-219 or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto.

[0186] In a further aspect of the invention there is provided an shRNA comprising a sequence according to any of SEQ ID NOs: 81-84, 142-145, 184-187, 195-198, 208-211, and 216-219 or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, or a vector comprising one or more of said shRNAs.

[0187] In an expression vector, the polynucleotide sequence encoding a construct, in this case suitably encoding the therapeutic nucleic acid, is typically operably linked to a control sequence which is capable of providing for the expression of the coding sequence by the host cell. Such expression vectors can be used to express a construct.

[0188] The term “operably linked” refers to a juxtaposition wherein the components described are in a relationship permitting them to function in their intended manner. A control sequence “operably linked” to a coding sequence is ligated in such a way that expression of the coding sequence is achieved under conditions compatible with the control sequences. Multiple copies of the same or different polynucleotide may be introduced into the vector.

[0189] The expression vector may then be introduced into a suitable host cell. Thus, a construct can be produced by inserting a polynucleotide sequence encoding a construct, suitably comprising or encoding the therapeutic nucleic acid, into an expression vector, introducing the vector into a compatible bacterial host cell, and growing the host cell under conditions which bring about expression of the polynucleotide sequence. Therefore, a further aspect of the invention is a host cell comprising the vector described above.

[0190] The vector may be a viral vector. A viral vector may be a viral particle that can be used in gene therapy, i.e. a viral particle that comprises all the required functional elements to express a construct in a host cell after administration as a therapeutic. Suitably in such embodiments, the host cell may be a cardiac cell or a neuron, for example. The viral vector may comprise a viral genome which comprises an expression cassette. The expression cassette maycomprise or encode the therapeutic nucleic acid operably linked to a promoter. The promoter may be specific for certain cell types, such as cardiac cells or neurons.

[0191] Suitable viral vectors include a parvovirus, a retrovirus, a lentivirus, or a herpes simplex virus. The parvovirus may be an adeno-associated virus (AAV). Suitably the viral vector is a recombinant adeno-associated viral (AAV) vector or a lentiviral vector. Suitably, the viral vector is an AAV viral vector.

[0192] Pharmaceutical Composition

[0193] The invention also provides a pharmaceutical composition comprising the therapeutic nucleic acid, the conjugate, delivery particle, or the vector according to the invention.

[0194] Optionally, the therapeutic nucleic acid may be present in the pharmaceutical composition as a physiologically tolerated salt. Suitably, physiologically tolerated salts retain the desired biological activity of the therapeutic nucleic acid and do not impart undesired toxicological effects. For therapeutic nucleic acids, suitable examples of pharmaceutically acceptable salts include (a) salts formed with cations such as sodium, potassium, ammonium, magnesium, calcium, polyamines such as spermine and spermidine, etc; (b) acid addition salts formed with inorganic acids, for example hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid and the like; (c) salts formed with organic acids such as, for example, acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and the like; and (d) salts formed from elemental anions such as chlorine, bromine, and iodine.

[0195] Optionally the therapeutic nucleic acids within the composition may be chirally- controlled. Suitably therefore the composition may be regarded as a chirally controlled oligonucleotide composition.

[0196] In some embodiments, any of the pharmaceutical compositions according to the invention may further comprise a pharmaceutically acceptable excipient, carrier, diluent, buffer, stabiliser or other materials well known to those skilled in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient.

[0197] The pharmaceutical carrier or diluent may be, for example, an isotonic solution. The precise nature of the carrier or other material may depend on the route of administration, e.g. oral, intravenous, cutaneous or subcutaneous, nasal, intramuscular and intraperitoneal routes.Examples of suitable compositions and methods of administration are provided in Esseku and Adeyeye (2011) and Van den Mooter G. (2006). For example, solid oral forms may contain, together with the active substance, diluents, e.g. lactose, dextrose, saccharose, cellulose, corn starch or potato starch; lubricants, e.g. silica, talc, stearic acid, magnesium or calcium stearate, and / or polyethylene glycols; binding agents; e.g. starches, gum arabic, gelatin, methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone; disaggregating agents, e.g. starch, alginic acid, alginates or sodium starch glycolate; effervescing mixtures; dyestuffs; sweeteners; wetting agents, such as lecithin, polysorbates, laurylsulphates; and, in general, non-toxic and pharmacologically inactive substances used in pharmaceutical formulations. Such pharmaceutical preparations may be manufactured in known manner, for example, by means of mixing, granulating, tableting, sugar-coating, or film-coating processes.

[0198] Oral formulations include such normally employed excipients as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. These compositions take the form of solutions, suspensions, tablets, pills, capsules, sustained release formulations or powders and contain 10% to 95% of active ingredient, preferably 25% to 70%. Where the pharmaceutical composition is lyophilised, the lyophilised material may be reconstituted prior to administration, e.g. a suspension. Reconstitution is suitably effected in buffer.

[0199] Capsules, tablets and pills for oral administration to an individual may be provided with an enteric coating comprising, for example, Eudragit “S”, Eudragit “L”, cellulose acetate, cellulose acetate phthalate or hydroxypropylmethyl cellulose.

[0200] Liquid dispersions for oral administration may be syrups, emulsions or suspensions. The syrups may contain as carriers, for example, saccharose or saccharose with glycerine and / or mannitol and / or sorbitol.

[0201] Suspensions and emulsions may contain as carrier, for example a natural gum, agar, sodium alginate, pectin, methylcellulose, carboxymethylcellulose, or polyvinyl alcohol. The suspensions or solutions for intramuscular injections may contain, together with the active substance, a pharmaceutically acceptable carrier, e.g. sterile water, olive oil, ethyl oleate, glycols, e.g. propylene glycol, and if desired, a suitable amount of lidocaine hydrochloride.

[0202] Solutions for intravenous administration or infusion may contain as carrier, for example, sterile water or suitably they may be in the form of sterile, aqueous, isotonic saline solutions.

[0203] For suppositories, traditional binders and carriers may include, for example, polyalkylene glycols or triglycerides; such suppositories may be formed from mixtures containing the active ingredient in the range of 0.5% to 10%, preferably 1% to 2%.

[0204] In some embodiments, the pharmaceutical composition may be formulated for intravenous, arterial, intradermal, intramuscular, intrathecal, intraperitoneal, subcutaneous, sublingual or oral administration. In one embodiment, the pharmaceutical composition is formulated for intravenous or intrathecal administration.

[0205] Pharmaceutical compositions such as those of the invention may comprise naked nucleic acid sequences or may comprise nucleic acid sequences in combination with antibodies, peptides, lipids, polymers or targeting systems. They may be delivered by any available technique. For example, the pharmaceutical composition may be introduced by needle injection, suitably intradermally, subcutaneously or intramuscularly. Alternatively, the pharmaceutical composition may be delivered directly across the skin using a delivery device such as particle- mediated gene delivery. The pharmaceutical composition may be administered topically to the skin, or to mucosal surfaces for example by intranasal, oral, or intrarectal administration. The pharmaceutical composition may be administered intravenously, arterially, intradermally, intramuscularly, intraperitonealy, subcutaneously, sublingually or orally (by ingestion). The pharmaceutical composition may also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e.g., patches and pumps, or formulations, which provide for the extended, slow, or controlled release thereof.

[0206] In one embodiment, the pharmaceutical composition is administered intravenously.

[0207] In some embodiments, the pharmaceutical composition is formulated such that the therapeutic nucleic acid reaches a target tissue in therapeutic concentrations. Suitably the target tissue is a tissue involved in the diseases to be treated. Suitably a target tissue is a tissue in which the pathogenic allele is typically expressed. In the case of the preferred diseases herein, suitably the target tissue is the heart, or the central nervous system (CNS).

[0208] Uptake of therapeutic oligonucleotides into a target tissue may be enhanced by several known transfection techniques, for example those including the use of transfection agents. Suitably therefore the pharmaceutical composition may comprise a transection agent. Examples of these agents include cationic agents, for example, calcium phosphate and DEAE- Dextran and lipofectants, for example, lipofectamine and transfectam.

[0209] Administration is typically in a "prophylactically effective amount" or a "therapeutically effective amount" (as the case may be, although prophylaxis may be considered therapy), this being sufficient to show benefit to the individual, e.g. an effective amount to prevent or delay onset of the disease or condition, to ameliorate one or more symptoms, to induce or prolong remission, or to delay relapse or recurrence.

[0210] The dose may be determined according to various parameters, especially according to the substance used; the age, weight and condition of the individual to be treated; the route of administration; and the required regimen. A physician will be able to determine the required route of administration and dosage for any particular individual. A typical daily dose of an active ingredient is from about 0.1 to 50 mg per kg of body weight dependent on the conditions mentioned above. The dose may be provided as a single dose or may be provided as multiple doses, for example taken at regular intervals, for example suitably, the dose may be administered daily, once every 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14 days, once every 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12 weeks, or once every 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months. Suitably the dose may be administered hourly. Suitably, the dose may be administered as two, three, four, five, six or more sub-doses separately at appropriate intervals throughout the day, optionally, in unit dosage forms. Typically therapeutic nucleic acids are administered at a dose of about 10-200 mg, about 10-190 mg, about 10-180 mg, about 10-170 mg, about 10-160 mg, about 10-150 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, or about 168 mg. Optionally smaller doses may be used in the range of 1 pg to 1 mg, suitably 1 pg to 10 μg of nucleic acid for particle mediated delivery and 10 μg to 1 mg for other routes.

[0211] Examples of the techniques and protocols mentioned above can be found in Remington's Pharmaceutical Sciences, 20th Edition, 2000, pub. Lippincott, Williams & Wilkins.

[0212] In some embodiments, the pharmaceutical composition comprises a further therapeutic agent. The further therapeutic agent may be any further agent which modulates the expression of the pathogenic allele, or any additional therapeutic or protective agent as described below, suitably which may be used for treating the dominant negative or gain-of function genetic disorder. The further therapeutic agent may be a further therapeutic nucleic acid, such as an antisense oligonucleotide. Optionally, the pharmaceutical composition may comprise two or more different therapeutic nucleic acids. Optionally, the pharmaceutical composition mayfurther comprise one or more therapeutic nucleic acids targeting different indel alleles. Optionally different indel alleles associated with the same pathogenic allele or different pathogenic alleles. Suitably combinations of any of the therapeutic nucleic acids described herein are envisaged. For example, a pharmaceutical composition may comprise a therapeutic oligonucleotide which is capable of binding to a target indel allele associated with a pathogenic TNNT2 allele, and / or a therapeutic oligonucleotide which is capable of binding to a target indel allele associated with a pathogenic TNNI3 allele, and / or a therapeutic oligonucleotide which is capable of binding to a target indel allele associated with a pathogenic MHY7 allele, and / or a therapeutic oligonucleotide which is capable of binding to a target indel allele associated with a pathogenic HTT allele.

[0213] Optionally, the one or more further therapeutic nucleic acids may be joined together and / or joined to the therapeutic nucleic acids of the first aspect or the ninth aspect.

[0214] Treatment

[0215] The invention provides a method of treating or preventing a dominant negative genetic disease or a gain-of-function genetic disease in a subject, the method comprising administering to the subject an effective amount of a therapeutic nucleic acid. The invention equally provides a therapeutic nucleic acid for use in the treatment or prevention of a dominant negative genetic disease or a gain-of-function genetic disease in a subject. Optionally for use in a method of treatment or prevention in a subject, the method optionally comprising administration of an effective amount of the therapeutic nucleic acid to the subject.

[0216] Treatment may comprise prevention, not necessarily cure of such diseases. Treatment may therefore comprise a lessening of any symptoms, delay of disease progression, and / or improvement of any symptoms.

[0217] Administration of the treatment is in an "effective amount" which may be considered as a “prophylactically effective amount” or a "therapeutically effective amount" (as the case may be, although prophylaxis may be considered therapy), this being sufficient to show benefit to the individual, e.g. an effective amount to prevent or delay onset of the disease or condition, to ameliorate one or more symptoms, to induce or prolong remission, or to delay relapse or recurrence. Suitable doses for administration are provided hereinabove.

[0218] For example, in relation to cardiomyopathies, treatment may slow the progression of disease. In some embodiments, wherein treatment is commenced in an early stage cardiomyopathy, treatment may delay the onset of middle and / or late stage cardiomyopathy. Insome embodiments, wherein treatment is commenced during middle stage cardiomyopathy, treatment may delay the onset of late stage cardiomyopathy.

[0219] In some embodiments, the present disclosure pertains to: a method of ameliorating, reducing the severity of, or slowing the onset or progression of a dominant negative genetic disease or a gain-of-function genetic disease.

[0220] Specifically, treatment of cardiomyopathies with the therapeutic nucleic acid of the invention improves heart function, suitably by normalizing contractility, wall thickness, cavity size, energetics, heart rhythm, and tissue composition, for example, and improves exercise capacity and symptom burden, for example as measured by peak oxygen consumption, 6 minute walk test, and patient reported outcome measures such as KCCQ score. Suitably, treatment of cardiomyopathies with the therapeutic nucleic acid prevents development of the disease in carriers of pathogenic alleles who do not yet display any signs of the cardiomyopathy, or lessens progression and / or promotes improvement in the case of those who display signs of cardiomyopathy.

[0221] In one embodiment, treatment of HCM with the therapeutic nucleic acid of the invention may have one or more of the following effects: prevents or lessens hypercontractility and left ventricular hypertrophy, improves myocardial oxygenation and energetic status, increases ventricular cavity size and diastolic filling, reduces levels of circulating biomarkers that indicate cardiac impairment (such as NT-proBNP), improves exercise capacity, and lowers symptom burden.

[0222] In one embodiment, treatment of DCM with the therapeutic nucleic acid of the invention may have one or more of the following effects: prevents or lessens impaired systolic contractility and ventricular ejection fraction, reduces ventricular cavity size, reduces levels of circulating biomarkers that indicate cardiac impairment (such as NT-proBNP), improves exercise capacity and lowers symptom burden.

[0223] Suitably, treatment with a therapeutic nucleic acid of the invention reduces expression of the pathogenic allele. Suitably, the expression of a disease-causing (pathogenic) allele may be inhibited compared to the expression of said pathogenic allele without treatment with the therapeutic nucleic acid. Suitably treatment with a therapeutic nucleic acid of the invention reduces expression of the pathogenic allele by at least 5%, suitably by at least 10%, suitably by at least 15%, suitably by at least 20%, suitably by at least 25%, suitably by at least 30%. Suitablytreatment with a therapeutic nucleic acid of the invention reduces expression of the pathogenic allele between 20-30%.

[0224] Suitably the therapeutic nucleic acid is capable of binding to an indel allele associated with the pathogenic allele present in mRNA or DNA, suitably in pre-mRNA. Suitably the therapeutic nucleic acid can invade DNA which suitably contains the target indel allele and induce transcriptional gene silencing by blocking the transcriptional machinery. Alternatively, the therapeutic nucleic acid can direct nuclease enzymes such as CAS9 to a target indel allele in the DNA where an edit or modification is intended to occur. Suitably, the therapeutic nucleic acid can direct an RNA-editing enzyme such as Cas13 to a target indel allele in the mRNA or pre-mRNA to cleave the mRNA or pre-mRNA and thereby reduce the expression of the pathogenic allele. Suitably the therapeutic nucleic acid can direct enzymes such as RNAse H to mRNA ore pre-mRNA and reduce the expression of the pathogenic allele by post-transcriptional silencing. Suitably its expression is reduced by inducing destruction of the pathogenic allele transcript, suitably of the pre-mRNA or mRNA transcript of the pathogenic allele, which suitably contains the associated allele of the target indel allele. Suitably destruction of the pathogenic allele transcript may be mediated by RNase H cleavage. In some embodiments, the present disclosure pertains to: a method of reducing the expression of a pathogenic allele in a subject, suitably comprising administering the therapeutic nucleic acid of the invention to the subject. Alternatively, the present disclosure pertains to a therapeutic nucleic acid for use in a method of reducing the expression of a pathogenic allele in a subject. In some embodiments, the present disclosure pertains to: a method of causing degradation of a pathogenic allele transcript in a subject, suitably comprising administering the therapeutic nucleic acid of the invention to the subject. Alternatively, the present disclosure pertains to a therapeutic nucleic acid for use in a method of causing degradation of a pathogenic allele transcript in a subject.

[0225] Suitably the therapeutic nucleic acid selectively lowers production of the pathogenic protein, suitably while retaining sufficient levels of wild type protein. In some embodiments, the therapeutic nucleic acid can reduce the level, expression and / or activity of transcripts of the pathogenic allele and / or pathogenic protein products thereof more than those of wild type transcripts and / or protein products thereof, in individual subjects and / or populations of subjects suitably who have a dominant negative genetic disorder or a gain-of-function genetic disorder. Suitably therefore the present disclosure provides therapeutic nucleic acids which are capable of allele-specific knockdown of a pathogenic allele transcript, wherein allele-specificknockdown (also referenced as allele-specific suppression, allele-selective approach, allele- selective silencing, or the like) preferentially decreases the level, expression and / or activity of a pathogenic allele transcript and / or a product thereof (e.g., protein) relative to the wild-type allele transcript and / or a product thereof (e.g., protein) (e.g., the expression, level and / or activity of wild-type protein is not significantly decreased, not decreased, remains the same, or increases). Suitably by at least 5%, suitably by at least 10%, suitably by at least 15%, suitably by at least 20%, suitably by at least 25%, suitably by at least 30%.

[0226] In some embodiments, the present disclosure relates to a method of reducing the level, expression and / or activity of transcripts of a pathogenic allele, suitably in a subject. Alternatively, the present disclosure relates to the therapeutic nucleic acid for use in a method of reducing the level, expression and / or activity of transcripts of a pathogenic allele, suitably in a subject. In some embodiments, the present disclosure relates to a method of reducing the level, expression and / or activity of a pathogenic protein, suitably in a subject. Alternatively, the present disclosure relates to the therapeutic nucleic acid for use in a method of reducing the level, expression and / or activity of a pathogenic protein, suitably in a subject. In some embodiments, the present disclosure pertains to: a method of allele-specific knockdown of a pathogenic allele transcript in a subject. Alternatively, the present disclosure relates to the therapeutic nucleic acid for use in a method of allele-specific knockdown of a pathogenic allele transcript in a subject. In some embodiments, the present disclosure pertains to: a method of allele-specific silencing of a pathogenic allele transcript in a subject. Alternatively, the present disclosure relates to the therapeutic nucleic acid for use in a method of allele-specific silencing of a pathogenic allele transcript in a subject.

[0227] Suitably therefore, a therapeutic nucleic acid of the invention reduces the abundance of mRNA of the pathogenic allele, suitably by targeting an allele of an indel comprised within said mRNA. Suitably therefore, a therapeutic nucleic acid of the invention reduces the abundance of protein produced from the pathogenic allele. In some embodiments, treatment may increase the abundance of mRNA of the wild type allele, suitably comprising a sequence as set forth in SEQ ID 220 (TNNT2 ENSG00000118194), SEQ ID 221 (MYH7 ENSG00000092054), SEQ ID 222 (TNNI3 ENSG00000129991), or SEQ ID 223 (HTT ENSG00000197386) a wild type protein comprising a sequence as set forth in SEQ ID 224 (TNNT2 ENSP00000499593.1), SEQ ID 225 (MYH7 ENSP00000347507.3), SEQ ID 226 (TNNI3 ENSP00000341838.5) or SEQ ID 227 (HTT ENSP00000347184.5) or a variant thereof. In some embodiments, treatment mayincrease the relative abundance of mRNA of the wild type allele, compared to mRNA of the pathogenic allele, suitably wherein the wild type mRNA comprises a sequence as set forth in SEQ ID 220 (TNNT2 ENSG00000118194), SEQ ID 221 (MYH7 ENSG00000092054), SEQ ID 222 (TNNI3 ENSG00000129991), or SEQ ID 223 (HTT ENSG00000197386) of a wild type protein compared to pathogenic protein, wherein the wild type protein comprises a sequence as set forth in SEQ ID 224 (TNNT2 ENSP00000499593.1), SEQ ID 225 (MYH7 ENSP00000347507.3), SEQ ID 226 ENSP00000341838.5) or SEQ ID 227 (HTT ENSP00000347184.5).

[0228] Suitably therefore, a therapeutic nucleic acid of the invention reduces the abundance of mRNA of the pathogenic allele, suitably by targeting an allele of an indel comprised within said mRNA. Suitably therefore, a therapeutic nucleic acid of the invention reduces the abundance of protein produced from the pathogenic allele. In some embodiments, treatment may increase the abundance of mRNA of the wild type allele, suitably comprising a sequence as set forth in SEQ ID 223 (HTT ENSG00000197386).

[0229] In some embodiments, the present disclosure pertains to: a method of reducing the abundance of mRNA encoding a pathogenic allele, suitably in a subject. Alternatively, the present disclosure relates to a therapeutic nucleic acid for use in a method of reducing the abundance of mRNA encoding a pathogenic allele in a subject. In some embodiments, the present disclosure pertains to: a method of reducing the abundance of protein produced from a pathogenic allele, suitably in a subject. Alternatively, the present disclosure relates to a therapeutic nucleic acid for use in a method of reducing the abundance of protein produced from a pathogenic allele in subject.

[0230] The relative abundance of mRNA may be determined by taking samples from the patient treated before treatment is commenced, and once treatment has begun, and measuring the abundance of mRNA by qPCR or RNA-sequencing. The relative abundance of protein may be determined by taking samples from the patient treated before treatment is commenced, and once treatment has begun, and measuring the abundance of protein by antibody based detection techniques.

[0231] In one embodiment, the therapeutic nucleic acid of the invention effectively silences the pathogenic allele. Suitably such that substantially only the wild type allele is expressed. In some embodiments, present disclosure relates to a method of silencing a pathogenic allele. Alternatively, the present disclosure relates to the therapeutic nucleic acid for use in a methodof silencing a pathogenic allele in a subject. Suitably any of the methods or uses in a method described herein, may be methods of treatment or prevention or uses for treatment or prevention of a dominant negative genetic disorder or a gain-of-function genetic disorder by causing any of the effects described above.

[0232] Suitably the therapeutic nucleic acid is efficacious in reducing the level, expression and / or activity of a pathogenic TNNT2 allele (or a protein product thereof), and is capable of mediating allele-specific knockdown of the pathogenic TNNT2 allele.

[0233] Suitably the therapeutic nucleic acid is efficacious in reducing the level, expression and / or activity of a pathogenic TNNI3 allele (or a protein product thereof), and is capable of mediating allele-specific knockdown of the pathogenic TNNI3 allele.

[0234] Suitably the therapeutic nucleic acid is efficacious in reducing the level, expression and / or activity of a pathogenic MHY7 allele (or a protein product thereof), and is capable of mediating allele-specific knockdown of the pathogenic MHY7 allele.

[0235] Suitably in other embodiments, therapeutic nucleic acid is efficacious in reducing the level, expression and / or activity of a pathogenic HTT allele (or a protein product thereof), and is capable of mediating allele-specific knockdown of the pathogenic HTT allele.

[0236] The treatment may further comprise administering an additional therapeutic agent to the subject. The additional therapeutic agent may be comprised within the pharmaceutical composition as described above, or may be separate. The additional therapeutic agent may be a neuroprotective agent, an anti-inflammatory agent, an agent that regulates inflammation or immune system, an antifibrotic agent, and / or an agent that protects cells from accumulation of misfolded protein or excitatory stimuli. Suitably the additional therapeutic agent may be administered at the same time, simultaneously with, the therapeutic nucleic acid or pharmaceutical composition thereof of the invention, or at a different time. Suitably the additional therapeutic agent may act synergistically with the therapeutic nucleic acid of the invention.

[0237] Subject

[0238] The present invention relates to treatment of a dominant negative genetic disorder or a gain-of-function genetic disorder by administering an effective amount of the therapeutic nucleic acid to a subject in need thereof.

[0239] Suitably the subject has a dominant negative genetic disorder or a gain-of-function genetic disorder. Suitably the subject has been diagnosed with a dominant negative genetic disorder or a gain-of-function genetic disorder. Suitable such disorders are listed above. In some embodiments, a subject is genetically verified to have such a disease prior to administration of the therapeutic nucleic acid of the invention.

[0240] Suitably the subject comprises a pathogenic allele which causes a dominant negative genetic disorder or a gain-of-function genetic disorder. Suitably the present invention treats subjects who are heterozygous for the pathogenic allele, suitably therefore who have one copy of a wild type allele and one copy of a pathogenic allele, which causes a dominant negative disorder or a gain-of-function disorder.

[0241] Suitably the subject comprises a pathogenic allele which is amenable to allele-specific knockdown. Suitably a pathogenic allele which is amenable to allele-specific knockdown is a pathogenic allele associated with i.e. located on the same chromosome, in phase with, or on the same haplotype as, an indel allele which is capable of being targeted by a therapeutic nuclei acid of the invention. Suitably the subject is heterozygous for the target indel allele. Suitably the subject comprises a pathogenic allele associated with a heterozygous target indel allele. Suitably at the genetic locus of the target indel allele, the subject having the dominant negative or gain- of function genetic disorder is heterozygous. Suitably therefore the indel allele is not associated with the wild type allele.

[0242] In some embodiments, the present disclosure provides for a method for determining the suitability of treatment of a subject for administration of a therapeutic nucleic acid of the invention, said method comprising the steps of: i) determining the presence of a pathogenic allele in a subject ii) determining whether the subject is heterozygous for a target indel allele to which a nucleic acid of the invention is capable of binding; iii) determining which allele of the target indel allele is associated with the pathogenic allele, and iv) determining that the subject is suitable for treatment if the subject has a pathogenic allele associated with a suitable heterozygous indel allele, wherein the subject is suffering from or at risk of developing a dominant negative genetic disorder or a gain-of-function genetic disorder, and iv) optionally administering an effective amount the therapeutic nucleic acid to the subject.

[0243] Suitably step (i) is determined by genetic analysis or genotyping of the subject. Suitably by known techniques. Suitably step (i) may comprise determining the presence of apathogenic TNNT2, TNNI3, MYH7, and / or HTT allele in the subject. Suitably step (i) may comprise determining the presence of a pathogenic HTT allele in the subject.

[0244] Various techniques can be used to determine if a particular indel allele is associated with, i.e. on the same chromosome, as a disease-associated sequence, such as a pathogenic allele. Typically, if the indel allele and the pathogenic allele are on the same chromosome, a therapeutic nucleic acid such as an ASO that targets the indel allele can also “target” the disease-associated pathogenic allele, thereby allowing a decrease in the expression, level and / or activity of the pathogenic allele.

[0245] Humans, among other living things, are diploid, and determining the linkage of alleles of genetic loci on the same or different chromosomes is desirable for phasing techniques. The sequences on corresponding chromosomes are known as haplotypes. The process of determining which alleles are on which chromosomes is known as phasing, haplotype phasing or haplotyping. Suitably therefore steps (ii) and (iii) may be carried out by phasing, suitably by a phasing assay. Phasing information is useful in patient stratification, forensics and various other applications in the treatment of genetic diseases. For additional general information about phasing, see, for example: Twehey et al. 2011 Nat. Rev. Genet. 12: 215-223; and Glusman et al. 2014 Genome Med. 6:73.

[0246] Phasing data can be important in allele-specific therapies for genetic diseases. In some diseases, a genetic lesion such as a deleterious missense mutation, repeat, deletion, insertion, inversion or other mutation has been identified. In some subjects, one allele of a gene can comprise a disease-associated mutation at a genetic locus, while the other allele is normal, wild- type or otherwise not or less disease-associated. In some embodiments, an allele-specific therapy can target a pathogenic allele comprising a disease-associated mutation, but not the corresponding wild-type allele. In some embodiments, an allele specific therapy can target a pathogenic allele comprising a disease-associated mutation at a particular locus, but not by directly targeting the locus, but rather by targeting a different locus on the pathogenic allele, suitably an indel allele. As a non-limiting example, an allele-specific therapy can target a pathogenic allele comprising a disease-associated mutation at a locus by targeting a different locus in the same allele, suitably an indel allele. As a non-limiting example, phasing data for an subject indicates if a particular indel allele is in phase (e.g., on the same chromosome or transcript) as the pathogenic allele and thus that indel allele can be targeted with a therapeutic nucleic acid.

[0247] One example of a phasing assay that can be suitably employed for detection of mutant allele in a patient is allele-specific PCR or allele-specific long-range PCR, such assay can be performed which employs allele-specific primers to detect mutations in nucleic acid sequences in the presence of wild-type variants of the sequences. Allele-specific PCR is a technique in which the variant of the nucleic acid sequence present in the PCR reaction mixture is selectively amplified and detected. Allele-specific PCR employs at least one "allele-specific primer." Tire tenn "allele-specific" primer generally refers to a primer whose extension occurs in a PCR reaction only when a specific variant of a nucleic acid sequence is present in the reaction mixture. In other words, allele -specific primers are designed in such a way that they discriminate between variants of nucleic acids and selectively multiply nucleic acid templates that include a variant to be detected.

[0248] Other means for indicating if a particular indel allele is in phase (e.g., on the same chromosome or transcript) as the pathogenic allele are haplotype assembly from long read sequencing such as Nanopore or PacBio sequencing, as described in Martin et al. (2022) and McElwain et al. (2023) , or PCR amplification of the sequence including both the pathogenic alle and the associated target indel allele, and subcloning of the products prior to Sanger sequencing, as described in Peters et al. (2014).

[0249] Suitably, the subject is mammalian. Suitably the subject is human. Suitably the subject may be male or female.

[0250] Suitably, the subject is any age. However, suitably the subject is between the ages of 1 month old to 100 years old, suitably between the ages of 1 years old and 90 years old, suitably between the ages of 2 years old to 80 years old, suitably between the ages of 4 years old to 75 years old.

[0251] The invention will now be described in relation to the following figures. FIGURES Figure 1. Single allele sequencing of rs377373012 heterozygous iPSC line with heterozygous TNNI3 R145G variant. (A) R145G sequence of R145 region on same strand as (B) showing insertion allele (duplication of AGAC) in rs377373012. (C) WT sequence of R145 region on same strand as (D) showing deletion allele of rs377373012.Figure 2. Sanger sequencing traces of TNNI3 rs377373012 indel CRISPR-Cas9 editing. Deletion allele, heterozygous allele (HET), and insertion allele sanger sequencing traces from single cell clones. Figure 3. Therapeutic Nucleic Acids are more allele-specific when targeting indels than when targeting SNPs, illustrated for targeting of MYH7 in HEK293 cells. A) Oligonucleotide gapmers were cotransfected along with constructs carrying either variant of rs2239578 (SNP) or rs34598192 (indel). B) Gapmers targeting variant of the SNP rs2239578 A / G are not able to achieve allele specific silencing according to the fluorescence intensity of HEK293 cells measured 48h after cotransfection using lipofectamine with 2 plasmids expressing MYH7, each one containing one the variants of the SNP rs2239578. C) Oligonucleotide gapmers targeting variant of the indel rs34598192 – / C are able to achieve allele specific silencing according to the fluorescence intensity of HEK293 cells measured 48h after cotransfection using lipofectamine with 2 plasmids expressing MYH7, each one containing one the variants of the indel rs34598192 – / C. Results were normalised to control samples transfected with a negative control oligo (at the same concentration) that does not target any sequences in the plasmid or endogenous targets in the cells. Data are presented as mean ± SEM for each oligo concentration. Figure 4. Allele-specific targeting of TNNI3 rs377373012 insertion C / CCTGT in HEK293 cells. Normalised fluorescence intensity of HEK293 cells 48 hours after cotransfection with plasmids expressing a different variant of TNNI3 rs377373012 indel C / CCTGT. Oligonucleotides SEQ ID 87 (E3), SEQ ID 92 (E8), SEQ ID 93 (E9), SEQ ID 94 (E10), SEQ ID 95 (E11), SEQ ID 101 (E17), SEQ ID 103 (E19) targeting the insertion were transfected at concentrations of 25nM, 50nM and 100nM showcases the ability of these gapmers to silence specifically the target allele. The results were normalised to control samples transfected with a negative control oligo at the same concentrations, which does not target any sequences in the plasmid or endogenous targets in the cells. Data are presented as mean ± SEM for each oligo concentration. Figure 5. Analysis of CLCN5 Expression in HEK293 Cells After Transfection with Oligonucleotides. CLCN5 qPCR relative to BACTIN levels in HEK293 cells 48 hours post- transfection with various concentrations of oligonucleotides using Lipofectamine. Control treatments, SEQ ID 180 (CTL16) and SEQ ID 181 (CTL20) at 100nM show baseline CLCN5 expression. Treatment with SEQ ID 87 (E3) at different concentrations (25nM, 50nM, and 100nM) results in a significant reduction of (chloride voltage-gated channel 5) CLCN5expression, indicating a potential off-target effect. In contrast, the new designs of gapmer oligonucleotides SEQ ID 101 (E17) and SEQ ID 103 (E19) at varying concentrations (25nM, 50nM, and 100nM) show no significant reduction in CLCN5 expression, suggesting that these designs are able to avoid the cleavage of the off-target CLCN5. Error bars represent the standard deviation from the mean of biological replicates. Figure 6. TNNI3 expression after E3 treatment of iPSC Cardiomyocytes with Homozygous and Heterozygous rs377373012 SNP. (A) TNNI3 gene expression after 72 hour treatment of E3 at 1µM and 10µM in Media on homozygous deletion, heterozygous, and homozygous insertion variants of rs377373012 SNP. Kruskal-Wallis test (ns=non-significant, * = p<0.05, ** = p<0.01). Figure 7. NPPB expression after E3 treatment of iPSC Cardiomyocytes with Heterozygous rs377373012 Insertion Allele in cis with Heterozygous R145G HCM variant. (A) NPPB gene expression after 72 hour treatment of E3 at 1µM and 10µM in media on R145G iPSC cardiomyocytes. One-way ANOVA test (ns=non-significant, ** = p<0.01). Figure 8. SNP-directed ASOs lowers Tm but minimally disrupts ASO:RNA heteroduplex. (A) On-target differential scanning fluorimetry (DSF) data (A:U pair). Shape: Two-state transitions, clear Tms. (B) Off-target DSF data (A-C mismatch). Shape: Two-state transitions, clear Tms. Figure 9. Indel-directed ASOs significantly disrupts ASO:RNA heteroduplex. (A) On-target DSF data (no bulge). Shape: Two-state transitions, clear Tms. (B) Off-target DSF data (4-nt bulge in ASO). Shape: (Usually) multistep transitions. Figure 10. SNP-directed ASO shows minimal to no selectivity in vitro. DSF data for MYH7 SNP-directed ASO 6G. (A) On-target DSF data (G:C pair). (B) Off-target DSF data (G:U wobble). Figure 11. TNNI3 indel-directed ASOs show selectivity in vitro. DSF data for TNNI3 INS- directed ASO E3. (A) On-target DSF data (no bulge). (B) Off-target DSF data (4-nt bulge in ASO). Figure 12. TNNI3 indel-directed ASOs show selectivity in vitro. psiCHECK-2 data from 293T cells treated with selected ASOs. Data are means ± SD (n=3). (A) On-target plasmid. (B) Off-target plasmid.Figure 13. TNNI3 indel-directed ASOs show selectivity in vitro. Gymnosis in iPS-derived cardiomyocytes treated with TNNI3 INS targeting ASOs E3 (A), E17 (B), or E6 (C). All doses are in µM. Figure 14. RNase H cleaves indel-directed ASO:RNA heteroduplexes. RNase H cleavage assay data from the experiment described in Example 4. Figure 15. MYH7 indel-directed ASOs are active in vitro. QuantiGene data from the experiments described in Example 5. (A) Three-point dose response experiments using a non-targeting control (NTC) ASO, S17 from Anderson et al. (2020), and ASOs C1-C6. (B) Seven-point dose response experiments using S17 and C1. All data are means ± SD (n=3). Figure 16. Sanger sequencing traces of MYH7 rs34598192 indel CRISPR-Cas9 editing. Deletion allele, heterozygous allele (HET), and insertion allele sanger sequencing traces from single cell clones. Figure 17. Sanger sequencing traces of TNNT2 rs45533739 indel CRISPR-Cas9 editing. Deletion allele, heterozygous allele (HET), and insertion allele sanger sequencing traces from single cell clones. Figure 18. Lack of selectivity in targeting intronic SNPs by MYH7 SNP-directed ASO 6G. RT- qPCR data from iPS-cardiomyocytes treated with MYH7 SNP-directed ASO 6G. Left: on-target (hom G). Right: off-target (hom A). All doses are in nM. Figure 19. CRISPR editing is more selective at an indel than at a SNP. Sanger sequencing data from edited iPSCs. (A) Cells edited using a gRNA targeting the insertion indel at rs377373012 in cells which are homozygous for either the deletion allele or the insertion allele. (B) Cells edited using a gRNA targeting the ‘C’ SNP at rs2754163 in cells which are homozygous for either ‘T’ or ‘C’. Figure 20. MYH7 indel-directed ASOs show selectivity. Gymnosis in iPS-derived cardiomyocytes treated with MYH7 deletion targeting ASOs C1 (A) or C4 (B). All doses are in µM. Figure 21. List of indels. Column ‘gene id’ refers to Ensembl IDs for human genome GRCh38.p14. Column ‘phenotype mim number’ refers to the Online Mendelian Inheritance in Man (OMIM) phenotype numbers associated with the gene. Column ‘genome het fract’ shows the fraction of heterozygous genotypes for the variant. Column ‘genome_variant_id’ shows the chromosome number, chromosome location, reference sequence and indel sequence, in this order. For example, variant ID ‘16-70265687-C-CA’ has a reference sequence ‘C’ on chromosome 16 at location 70265687 whereas the indel has a ‘CA’ at this position. Column ‘genome flags’ refers to annotation flags from the genome variant database, which may indicate features such as low complexity regions, or quality filters. Column ‘lof.oe ci.upper’ refers to the upper bound of the confidence interval for theobserved / expected ratio of predicted loss of function variants in the gene, as reported by gnomAD. Column ‘odisea-tissue’ shows the tissue or tissues most affected by the disease associated with the gene, as reported in the ODISEA dataset.

[0252] EXAMPLES

[0253] The invention will now be demonstrated by way of the following non-limiting examples.

[0254] Materials and Methods

[0255] iPSC lines and culture. The iPSC line KOLF2-C1 (WTSIi018-B-1) (Wellcome Sanger Institute) was cultured in E8 Flex (Thermo Fisher Scientific, A2858501) on Geltrex (Thermo Fisher Scientific, A1413302), passaging at 60-80% confluence. The iPSC line “R145G” was derived from a patient with a TNNI3 R145G variant. Previously, dermal fibroblasts were collected and reprogrammed with a Sendai virus containing Oct3 / 4–Sox2, cMyc, and Klf4. This iPSC line was cultured in StemMACS (Miltenyi Biotec, 130-104-368) on Geltrex, passaging at 60-80% confluence.

[0256] CRISPR-Cas9 genome editing. The insertion allele of indel rs377373012 was introduced by CRISPR-Cas9 genetic editing of the KOLF2-C1 iPSC line. One million KOLF2- C1 cells were electroporated with a cas9 nuclease (IDT, 1081060) using electroporated using a Lonza 4D-Nucleofector (Lonza) (protocol CA137, P4 kit (Lonza, V4XP-4012)) using the guide RNA (SEQ ID 228) (CCAGGGGUCCCUUAGGAGAC) and HDR template (SEQ ID 229) (GCGCGGTCCCCCGACCTCTTGTTCAGAGGGGACTCCAGGGGTCCCTTAGGAGACAG ACAGGACACAGCCCACCACTAACCCCCCTCCTTGGTTTCTCTCCTTCCA). Single cell clones were isolated, expanded, and editing confirmed by Sanger sequencing of extracted DNA. The pathogenic TNNI3 R145G variant was removed by CRISPR-Cas9 genetic editing of the R145G iPSC line, in the same way with the guide RNA (SEQ ID 230) (AGAGAUCCUCACUCUCCCCA) and HDR template (SEQ ID 231) (AGCGCCTGCATCATGGCATCTGCAGAGATCCTCACTCTCCGGAGGGTGGGCCGCTT AAACTTGCCTCGAAGGTCAAAGATC).

[0257] The insertion allele of indel rs34598192 was introduced by CRISPR-Cas9 genetic editing of the KOLF2-C1 iPSC line. One million KOLF2-C1 cells were electroporated with a cas9 nuclease (IDT, 1081060) using a Lonza 4D-Nucleofector (Lonza) (protocol CA137, P4 kit (Lonza, V4XP-4012)) using the guide RNA (AAGGGAAGUGAUUUGAUGCA, SEQ ID NO: 237) and HDR template(CTAGACTCGGGCCACCTGGCCCAAGCAAGGAGCACACTGACTAGCCTTGCATCAAC ATCACTTCCCTTCCCAGCAATAAGGCTGGCTGTGGACCAACAGTTCTCCA, SEQ ID NO: 238). Single cell clones were isolated, expanded, and editing confirmed by Sanger sequencing of extracted DNA.

[0258] The deletion allele of indel rs45533739 was introduced by CRISPR-Cas9 genetic editing of the KOLF2-C1 iPSC line. One million KOLF2-C1 cells were electroporated with a cas9 nuclease (IDT, 1081060) using a Lonza 4D-Nucleofector (Lonza) (protocol CA137, P4 kit (Lonza, V4XP-4012)) using the guide RNA (UCUGCUCAGAAGAGAAGUCC, SEQ ID NO: 239) and HDR template (TCTCCACCTGCCTGAGGCACATACCTTCAACAGCTGCTTCTGCTCAGAAGTCCAGGC AGCAAGAGAAGAGAGAAGAGGTGGGTCAGTTTCGAACCAGGCT, SEQ ID NO: 240). Single cell clones were isolated, expanded, and editing confirmed by Sanger sequencing of extracted DNA.

[0259] Cardiomyocyte differentiation. Methods adapted from Lian et al. (2020) and Burridge et al. (2014) At 90% confluence, iPSC cells were differentiated using 8 to 12 μM CHIR (Selleck Chemicals, S2924-SEL-25mg) in N21 (Bio-Techne Ltd, AR010) or B27 (Thermo Fisher Scientific, 17504044) in RPMI 1640 (Thermo Fisher Scientific, 21875034) without insulin for 24 hours, diluted to half concentration for a further 24 hours, then changed to N21 or B27 in RPMI 1640 without insulin for another 24 hours. 5 μM IWP2 (Cambridge Bioscience, I9060- 5mg) or IWR1 (Selleck Chemicals, S7086-SEL-10mg) in N21 or B27 in RPMI 1640 without insulin was added for 48 hours then cultured for another 48 hours in N21 or B27 in RPMI 1640 without insulin. Media was replaced every 48 hours with N21 or B27 in RPMI 1640. Metabolic selection of cardiomyocytes was performed once beating was observed. Addition of 4 mM lactic acid (VWR, 0219022810) in N21 or B27 in glucose-free RPMI 1640 (Thermo Fisher Scientific, 11879020) for two 48-hour cultures. Or cardiomyocytes were differentiated from iPSC cells using 6 μM CHIR for 24 hours, diluted to 3 μM CHIR for a further 24 hours then cultured in 2 μM C-59 (APExBIO, A8685) for 48 hours all in RPMI 1640 (500 μg / ml recombinant human albumin and 213 μg / ml L-ascorbic acid 2-phosphate) then cultured in RPMI 1640 supplemented with L-ascorbic acid 2-phosphate and recombinant human albumin.

[0260] Cardiomyocyte replating. Cardiomyocytes were passaged between day 17 and day 20 by removal of media, addition of 1 ml 10X TrypLE Select Enzyme (Thermo Fisher Scientific, A1217701) per well for 10 minutes at 37°C, then addition of 2 ml replating media (90 mlN21 / RPMI (Bio-Techne Ltd, AR010) or RPMI 1640 with 500 μg / ml recombinant human albumin and 213 μg / ml L-ascorbic acid 2-phosphate, 10 ml KOSR (Thermo Fisher Scientific, 10828010), 20 μl Thiazovivin (Selleck Chemicals, S1459-SEL-10mg)). Cells were resuspended and passed through a 100 μm cell strainer (Greiner Bio-One, 542000). Cardiomyocytes were counted, spun at 200g for 5 minutes, resuspended in replating media at 375,000 cells per ml, and plated into 24 well plates. Media was changed to N21 in RPMI 1640 or RPMI 1640 with 500 μg / ml recombinant human albumin and 213 μg / ml L-ascorbic acid 2-phosphate after 24 hours and replaced every 48 hours thereafter.

[0261] Cardiomyocyte Transfections and qPCR. Antisense oligonucleotides were added to the cardiomyocyte media without any transfection reagent for 72 hours then harvested. RNA was harvested by removal of media, two washes in PBS (Thermo Fisher Scientific, 14190094), then scraped in 300 μl of RLT with beta-mercaptoethanol (Merck, 63689-25ML-F). RNA was extracted using the Qiagen RNeasy kit (Qiagen, 74104) following the manufacturer’s instructions. RNA was reverse transcribed into cDNA using a High-Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific, 4368814). Quantitative PCR was performed using TaqMan probes for TNNI3 (Thermo Fisher Scientific, Hs00165957_m1), MYH7 (Thermo Fisher Scientific, Hs01110632_m1), NPPB (Thermo Fisher Scientific, Hs00173590_m1), beta-tubulin (Thermo Fisher Scientific, Hs00742828_s1), or GAPDH (Thermo Fisher Scientific, Hs02758991_g1) and TaqMan Fast Universal PCR Master Mix (Thermo Fisher Scientific, 4366073) on a QuantStudio 7 Flex Real-Time PCR System (Thermo Fisher Scientific, Applied Biosystems).

[0262] Generation and Characterization of iPSC Lines. iPSC lines heterozygous for the R145G variant and rs377373012 were generated using standard reprogramming techniques from patient-derived fibroblasts. Single allele sequencing was performed to confirm the presence of the R145G variant and the heterozygous state of rs377373012. Sequencing was carried out using Sanger sequencing with primers specific to the R145 region on the same strand. Both wild-type (WT) and mutant sequences were analyzed, ensuring no duplication in the WT sequence and the presence of duplication in the mutant sequence.

[0263] CRISPR-Cas9 Genome Editing. CRISPR-Cas9 editing was employed to generate specific alleles of rs377373012. Single cell clones were obtained, and Sanger sequencing was used to verify the editing outcomes. Sequencing traces were analyzed for deletion, heterozygous,and insertion alleles. CRISPR-Cas9 components were delivered to iPSCs using lipofectamine transfection.

[0264] Oligonucleotide Design and Transfection. Oligonucleotide synthesis was performed with a KA synthesiser using the phosphoramidite method on a solid support to facilitate the sequential addition of nucleotides. The procedure commenced with the attachment of the initial nucleotide to the solid support. Each subsequent nucleotide was added through a series of repetitive cycles involving four key steps: deprotection, coupling, capping, and oxidation. Deprotection involved the removal of the 5'-dimethoxytrityl (DMT) protective group, exposing the 5'-hydroxyl group for the next coupling reaction. The coupling step entailed the reaction of the deprotected nucleotide with the incoming phosphoramidite nucleotide, forming a phosphite triester linkage. Any unreacted hydroxyl groups were inactivated in the capping step to prevent the formation of truncated sequences. Oxidation then converted the phosphite triester linkage to a more stable phosphate triester. Upon completion of the desired nucleotide sequence, the synthesized oligonucleotide was cleaved from the solid support and subjected to deprotection to remove any remaining protective groups. Therapeutic nucleic acids (antisense oligonucleotide gapmers) were designed to target specific indels and SNPs as comparisons. We used the placement of the target recognition sequence of the ASO which binds to the indel allele with a preference for the target recognition sequence to begin at nucleotide 8 of the ASO sequence that can be displaced if needed to reduce off-target effects in other targets, reduction of the length of DNA in the central region of the ASO gapmer from a standard of 10 to as short as 6 or 7 nucleotides to increase allele-specificity, and fine-tuning of chemistries such as bridge nucleic acids (locked nucleic acids, constrained ethyl nucleotides) placed within the ASO sequence to maximize binding to the target of interest and minimize off-target effects in other genes and increase potency. Specific antisense oligonucleotide sequences that were optimised and used in the examples are provided below. Transfection was performed using Lipofectamine 3000 (Thermo Fisher Scientific), following the manufacturer’s protocol. HEK293 were cotransfected with plasmids expressing either variant of the indels or SNPs. Fluorescence intensity was measured 48 hours post-transfection to assess allele-specific silencing. Data were normalized to control samples transfected with a non-targeting oligo. After this high-throughput phase iPSC Cardiomyocytes were treated with the therapeutic nucleic acids without using any transfection reagent and at concentrations of 1μM and 10μM for 72 hours. All experiments were performed in biological triplicates.

[0265] Example 1

[0266] Results

[0267] Single Allele Sequencing of rs377373012 Heterozygous iPSC Line. Single allele sequencing was used to analyse the wild type (WT) and R145G sequences of the R145 region on the same strand. Figure 1C and D shows the WT sequence without duplication of rs377373012, while Figure 1A and B shows the R145G sequence with duplication of rs377373012, confirming the presence of the heterozygous R145G indel allele. Placement of the target recognition sequence in the oligonucleotide which binds to the indel allele was carefully analysed to ensure accurate allele characterisation.

[0268] Sanger Sequencing of CRISPR-Cas9 Edited Clones. Sanger sequencing traces of rs377373012 CRISPR-Cas9 edited single cell clones revealed deletion, heterozygous, and insertion alleles (Figure 2). This confirms the successful editing of rs377373012 in the iPSC lines, demonstrating the efficacy of the CRISPR-Cas9 technique in generating the desired indel allelic variants.

[0269] Sanger Sequencing of CRISPR-Cas9 Edited Clones. Sanger sequencing traces of rs34598192 CRISPR-Cas9 edited single cell clones revealed deletion, heterozygous, and insertion alleles (Figure 16). This confirms the successful editing of rs34598192 in the iPSC lines, demonstrating the efficacy of the CRISPR-Cas9 technique in generating the desired indel allelic variants.

[0270] Sanger Sequencing of CRISPR-Cas9 Edited Clones. Sanger sequencing traces of rs45533739 CRISPR-Cas9 edited single cell clones revealed deletion, heterozygous, and insertion alleles (Figure 17). This confirms the successful editing of rs45533739 in the iPSC lines, demonstrating the efficacy of the CRISPR-Cas9 technique in generating the desired indel allelic variants.

[0271] Allele Specificity of Antisense Oligonucleotide Gapmers Targeting Indels compared to Targeting SNPs. We successfully achieved allele-specific silencing across all targets by fine-tuning the placement of the variant in the binding region in the oligonucleotide, reducing the DNA in the central region of the oligonucleotide, and optimizing the chemistries to minimize off-target effects and increase potency. Antisense oligonucleotide gapmers targeting the indel MYH7 rs34598192 (– / C) showed higher allele specificity compared to gapmers targeting the commonly targeted SNP MYH7 rs2239578 (A / G). As shown in Figure 3A, gapmers targeting the indel achieved allele-specific silencing, indicated by greater normalizedfluorescence differences between constructs containing the insertion and deletion variants. In contrast, gapmers targeting the SNP did not achieve allele-specific silencing, as evidenced by the normalized fluorescence differences between constructs containing the two SNP variants (Figure 3B). These results highlight the effectiveness of our approach, particularly the strategic placement of the variant in the binding region of the oligonucleotide, and the reduction of the DNA in the central region of the oligonucleotide gapmer structure, in enhancing specificity.

[0272] Allele-Specific Targeting of TNNI3 rs377373012. Figure 4 demonstrates the normalized fluorescence intensity of HEK293 cells transfected with plasmids expressing different variants of TNNI3 rs377373012 (C / CCTGT). The antisense oligonucleotides E3 (SEQ ID 87), E8 (SEQ ID 92), E9 (SEQ ID 93), E10 (SEQ ID 94), E11 (SEQ ID 95), E17 (SEQ ID 101), and E19 (SEQ ID 103) demonstrated specific silencing of the target allele at various concentrations. This indicates effective allele-specific targeting of the oligonucleotides herein, further underscoring the precision of our designed oligonucleotide gapmers. By fine-tuning the chemistries and reducing the DNA in the gapmers, we minimized off-target effects and increased potency, as seen with the targeted silencing of the insertion variants.

[0273] Analysis of CLCN5 Expression. To assess off-target effects, we measured CLCN5 expression in HEK293 cells transfected with various concentrations of antisense oligonucleotides (25nM, 50nM, and 100nM) using Lipofectamine 3000. As shown in Figure 5, CLCN5 expression relative to BACTIN was significantly reduced following treatment with SEQ ID 87 (E3), indicating potential off-target effects. However, the optimized oligonucleotide gapmers SEQ ID 101 (E17) and SEQ ID 103 (E19) did not significantly reduce CLCN5 expression, demonstrating their efficacy in avoiding off-target effects. These results validate the strategic placement of the variant in the binding region of the oligonucleotide, reduction of the DNA in the central region of the gapmer, and fine-tuning of chemistries, which were crucial in minimizing off-target effects and enhancing the potency of our therapeutic nucleic acids.

[0274] TNNI3 Expression in iPSC Cardiomyocytes. Figure 6 shows TNNI3 gene expression after treatment with E3 oligonucleotides in iPSC cardiomyocytes with different variants of rs377373012. Treatment with E3 at 1µM and 10µM did not significantly affect TNNI3 expression in the homozygous deletion variant line. However, E3 at 10µM showed ~50% suppression of TNNI3 expression in the heterozygous line and greater than 50% suppression in the homozygous insertion variants. These results, determined by the Kruskal-Wallis test, highlight the allele- specific silencing potential of the E3 oligonucleotides. The placement of the variant in thebinding region of the oligonucleotide and the optimization of chemistries played a pivotal role in enhancing the efficacy and specificity of the treatment.

[0275] NPPB Expression in iPSC Cardiomyocytes. NPPB gene expression in iPSC cardiomyocytes heterozygous for rs377373012 and the R145G HCM variant after E3 treatment is shown in Figure 7. Untreated cells with the pathogenic R145G HCM variant exhibited elevated levels of NPPB, indicative of HCM pathophysiology. Treatment with E3 at 1µM and 10µM significantly normalized NPPB expression levels, as assessed by a one-way ANOVA test, indicating a rescue of the HCM phenotype. These findings suggest that the E3 oligonucleotide effectively mitigates the pathological expression of NPPB in HCM-affected cardiomyocytes, showcasing the impact of our design strategy on therapeutic efficacy of the therapeutic nucleic acids of the invention. (2023)

[0276] Example 2: INDEL is more disruptive to ASO:RNA heteroduplex than SNP

[0277] Materials and Methods

[0278] Methods were adapted from Stulz et al (2023). Reactions were assembled using 1 µL of 20 µM antisense oligonucleotide (ASO), 1 µL of 20 µM RNA target (Table 1), 0.05 µL Quant- iT RiboGreen (Life Technologies R11491), and 7.95 µL of 10 mM phosphate buffer with no added NaCl, pH 7.0, in technical triplicate in a 384 well plate. Samples were denatured by heating to 95 ^C and cooling to 20 ^C at ramp rates of -2.52 ^C s-1and -2.17 ^C s-1, respectively, and then fluorescence intensity was monitored over the temperature range 20 to 95 ^C at a data collection rate of 25 points per degree, on a Quantstudio 6 Pro (Thermo Fischer Scientific). Fluorescence intensity was plotted as a function of temperature and a melting temperature (Tm) was identified as the minimum of the first derivative of the melting curve.

[0279] Table 1. RNA targets used for differential scanning fluorimetry (DSF) studies.

[0280] Results

[0281] Figure 8 shows differential scanning fluorimetry (DSF) data for selected ASOs in the A series (A1-A6), which were designed to target the MYH7 SNP rs2239578 (U). These ASOs were paired with either their on-target RNA (left), where an A:U base pair is expected to form at the SNP site in the heteroduplex, or their off-target RNA (right), where an A-C mismatch is expected to form at the SNP site in the heteroduplex. In both sets of data, two-state transitions were observed and clear Tms were identified. The difference between the Tms (i.e., delta Tm) for the ASO:on-target RNA and ASO:off-target RNA pairs was, on average, approximately 6 ^C (Table 2).

[0282] Figure 9 shows DSF data for selected ASOs in the E series (E1-E6), which were designed to target the TNNI3 INDEL rs377373012 (INS). These ASOs were paired with either their on-target RNA (left), where a perfectly complementary (i.e., without mismatches or bulges) heteroduplex is expected to form, or their off-target RNA (right), where a four-nucleotide bulge in the ASO strand of the heteroduplex is expected to form. In only the on-target DSF data were two-state transitions reliably observed and therefore clear Tms identified; the melting curves for the ASO:off-target RNA pairs often displayed multistep transitions indicative of significantly disrupted duplexes.

[0283] Figure 10 shows DSF data for an MYH7 SNP rs2239578 (C)-directed ASO (6G, SEQ ID NO: 262) with mixed LNA / MOE wing chemistry paired with either its on-target RNA (left) or its off-target RNA (right); the delta Tm is approximately 7 ^C (Table 2). Figure 11 shows DSF data for a TNNI3 INDEL rs377373012 (INS)-directed ASO (E3) of the same chemistry paired with either its on-target RNA (left) or its off-target RNA (right); the delta Tm is approximately 22 ^C (Table 2), although the Tm for the ASO:off-target pair is only an estimate, as the melting curve displays a multistep transition indicative of a significantly disrupted duplex.

[0284] Taken together, these results indicate that an ASO and a mismatched INDEL target pair can create a more disrupted ASO:RNA heteroduplex than an ASO and a mismatched SNP target pair.

[0285] Table 2. Melting temperatures (Tms) determined by DSF for selected ASOs in the A, B, and E series, which were designed to target the MYH7 SNP rs2239578 (U), MYH7 SNP rs2239578 (C), and TNNI3 INDEL rs377373012 (INS), respectively. *Estimate only, as the melting curve for the ASO:off-target RNA pair displays a multistep transition indicative of a significantly disrupted duplex.

[0286] Example 3: Selectivity of targeting

[0287] Materials and Methods

[0288] Dual luciferase reporter assay (psiCHECK). Methods were adapted from Tran et al (2022). 293T (ATCC CRL-3216) cells were seeded in 96 well plates (Greiner Bio-One 655098) at a density of 10,000 cells / well. Cells were cultured in DMEM (Gibco 0566016) supplemented with GlutaMAX, 10% FBS, and antibiotics (Gibco 15240062). After 16 h, the cells were transfected with the psiCHECK-2 vector (Promega C8021) containing the relevant insert (Table 3) using Lipofectamine 3000 (ThermoFisher L30000008) according to the manufacturer’s instructions. Each well was transfected with 20 ng of the plasmid. The TNNI3 insertion plasmidwas prepared in-house by restriction digest using AsiSI and NotI (New England Biolabs R0630S and R0189S, respectively). All other plasmids were prepared by Integrated DNA Technologies (IDT) using an insertion site in the multiple cloning region of psiCHECK-2. At 24 h post-plasmid transfection, the cells were treated with the indicated ASO using Lipofectamine RNAiMAX (ThermoFisher 13778075) according to the manufacturer’s instructions. At 24 h post ASO- transfection, the cells were lysed and the activities of Firefly and Renilla luciferases were measured using the Dual-Luciferase Reporter 1000 Assay System (Promega E1980) as per the manufacturer’s instructions with the following modifications: 15 µL of Luciferase Assay Reagent II and 15 µL of Stop & Glo Reagent were used per well. Luminescence was recorded on a CLARIOstar plate reader. Data were analysed in Microsoft Excel (v16.93.1). Renilla luminescence was divided by Firefly luminescence to account for transfection efficiency. The Renilla / Firefly values for each ASO-treated well were then normalised to the mean Renilla / Firefly value for the untreated wells. Data were plotted in GraphPad Prism (v10.5.0). The data in the plots are mean normalised Renilla / Firefly signals ± standard deviations for two to twelve replicates.

[0289] Table 3. Inserts cloned into the psiCHECK-2 vector. The inserts in bold were used to generate the data presented in Figure 12.

[0290] Results

[0291] Figure 12 shows the activities of the ASOs in the E series, which were designed to target the TNNI3 INDEL rs377373012 (INS), at three doses on either an on-target plasmid (Figure 12A) or an off-target plasmid (Figure 12B). Many ASOs showed on-target activity; fewer showed off-target activity. Some ASOs were active on only the on-target plasmid, or more active on the on-target plasmid relative to the off-target plasmid, suggesting they are selective for their on-target RNA. For each ASO, an average selectivity index was calculated by dividing the residual off-target signal by the residual on-target signal at each dose and then averaging the selectivity index values of the three doses. Among the ASOs in the E series, E03 has a high average selectivity index of 2.0, E17 has a medium average selectivity index of 1.5, and E06 has a medium-to-low average selectivity index of 1.3. The results agree with the gymnosis data collected in iPS-derived cardiomyocytes with different variants of rs377373012 (Figure 13) and indicate that TNNI3 INDEL-directed ASOs are active and selective in vitro.

[0292] Figure 13 shows the activities of three ASOs in the E series on iPSC-derived cardiomyocytes, which were designed to target the TNNI3 INDEL rs377373012 (INS), at doses ranging from 0.5 micromolar to 20 micromolar either on a iPSC derived cardiomyocyte line that is homozygous for the target allele (left) or homozygous for the non-target allele (right).

[0293] Example 4: RNase H assay

[0294] Materials and Methods

[0295] Reactions were assembled using 0.5 µL of 10 µM ASO, 1 µL of 10 µM RNA (5’- GGUCCCUUAGGAGACAGACAGGACAC-3’, SEQ ID NO: 258), and 15 μL RNase H reaction buffer (New England Biolabs M0297S). Heteroduplexes were annealed by heating at 90 °C for 2 min and then cooling to 25 °C at a ramp rate of -0.1 °C s-1. Then, 0.1 µL RNase H (New England Biolabs M0297S) was added and reactions were brought to 20 µL with RNase-free water. Reactions were incubated at 37 °C for 20 min and then stopped by adding 1 µL of 0.5 M EDTA and 9 µL Gel Loading Dye, Purple (6X) (New England Biolabs B7024S), heating at 90 °C for 2 min, and chilling on ice. To resolve the cleavage products, 15 µL of each reaction were separated by 18% polyacrylamide gel electrophoresis (PAGE; Invitrogen HC2040). The marker was 14-30 ssRNA Ladder Marker (Takara Bio 3416). The gel was stained for 10 min in SYBR Gold Nucleic Acid Gel Stain (Thermo Fisher Scientific S11494) and imaged on a Bio-Rad ChemiDoc.

[0296] Results

[0297] Figure 14 shows that cleavage products corresponding to ~14 nt or shorter single- stranded RNAs are detected following the incubation of heteroduplexes formed between selected ASOs in the E series, which were designed to target the TNNI3 INDEL rs377373012 (INS), and their on-target RNA. The results indicate that TNNI3 INDEL-directed ASO:RNA heteroduplexes are cleaved by RNase H in vitro.

[0298] Example 5: QuantiGene Singleplex Assay

[0299] Materials and Methods

[0300] Methods were adapted from Anderson et al (2020). Immortalised human skeletal muscle myoblast cell lines (MRC CNMD Biobank London L954 / 1284 M-I) were seeded in 96 well plates (Corning 3598) at a density of 10,000 cells / well. Cells were cultured in Skeletal Muscle Cell Growth Medium (PromoCell C-23060) supplemented with 10% FBS and antibiotics (Gibco 15240062) until the cells reached 80% confluency. The cells were then washed with PBS, and Skeletal Muscle Differentiation Medium (PromoCell C-23061) supplemented with antibiotics (Gibco 15240062) was added. The cells were differentiated for five to seven days until differentiation was complete and myotubes were observed.

[0301] The cells were then treated under free uptake (gymnosis) conditions, i.e., in the absence of transfection reagents, with the indicated ASO. For three-point dose response experiments, the cells were treated for three days before being lysed. For seven-point dose response experiments, the cells were treated for 10 days, and media containing the ASO was replenished at day five. MYH7 (Assay ID: SA-10161) and PPIB (Assay ID: SA-10003) were quantified using the QuantiGene Singleplex Assay (ThermoFisher QS0013) as per the manufacturer’s instructions. Luminescence was recorded on a CLARIOstar plate reader. Data were analysed in Microsoft Excel (v16.93.1). The average luminescence of blank wells was subtracted from ASO-treated wells. The luminescence measured for MYH7 was divided by the luminescence measured for PPIB, and the MYH7 / PPIB values for each ASO-treated well were then normalised to the mean MYH7 / PPIB of the untreated wells. Data were plotted in GraphPad Prism (v10.5.0). The data in the plots are mean normalised MYH7 / PPIB signals ± standard deviations for three replicates.

[0302] Results

[0303] Figure 15A shows the activity of a previously published positive control compound S17 (AAAGgatgtaagaTGCA, SEQ ID NO: 259) as well as selected ASOs in the C series, which were designed to target the MYH7 INDEL rs34598192 (DEL), at three doses on the endogenousexpression of MYH7 in differentiated human skeletal muscle myoblasts. Figure 15B shows the activity of the positive control compound S17 and C01 at seven doses on the endogenous expression of MYH7 in differentiated human skeletal muscle myoblasts. The calculated absolute IC50 for S17 was approximately 38 nM, and the calculated absolute IC50 for C01 was approximately 3.3 µM. The results indicate that MYH7 INDEL-directed ASOs are active in vitro.

[0304] Example 6: Lack of selectivity in targeting intronic SNPs

[0305] Figure 18 shows lipofection of an MYH7 SNP rs2239578 (C)-directed ASO into iPS- cardiomyocytes that are either homozygous for the G (target SNP) or for A (WT) at the MYH7 rs2239578 locus, showing a lack of selectivity of this ASO in targeting an intronic SNP in vitro.

[0306] Example 7: CRISPR

[0307] Materials and Methods

[0308] iPSC lines and culture. The iPSC line KOLF2-C1 (homozygous deletion for rs377373012, and homozygous ‘T’ for rs2754163 (WTSIi018-B-1)) (Wellcome Sanger Institute) was cultured in E8 Flex (Thermo Fisher Scientific, A2858501) on Geltrex (Thermo Fisher Scientific, A1413302), passaging at 60-80% confluence. The iPSC line “Hom insertion” for rs377373012 was derived from the KOLF2-C1 iPSC line by CRISPR-Cas9 genetic editing as described above (under Materials and Methods subheading ‘CRISPR-Cas9 genome editing’). The iPSC line “Hom C” for rs2754163 was derived from a patient where, previously, dermal fibroblasts were collected and reprogrammed with a Sendai virus containing Oct3 / 4–Sox2, cMyc, and Klf4. This iPSC line was cultured in StemMACS (Miltenyi Biotec, 130-104-368) on Geltrex, passaging at 60-80% confluence.

[0309] CRISPR-Cas9 genome editing. Guide RNAs targeting the insertion allele of rs377373012 (UGUGUCCUGUCUGUCUCCUA, SEQ ID NO: 260) and the ‘C’ allele of rs2754163 (AUUUCCCAGGGGCUUGAGAG, SEQ ID NO: 261) were electroporated with cas9 nuclease (IDT, 1081060) using a Lonza 4D-Nucleofector (Lonza) (protocol CA137, (Lonza, V4XP-4012)) into iPSC lines homozygous for either insertion or deletion allele of rs377373012 and homozygous for the ‘T’ or ‘C’ allele of rs2754163. Percentage editing was analysed after 72 hours by Sanger sequencing of extracted DNA and TIDE analysis (Brinkman et al. 2014).

[0310] Results

[0311] Figure 19A left graph shows percentage editing of the guide RNA targeting the insertion allele of rs377373012 in iPSC lines homozygous for the deletion allele or the insertionallele, the graph on the right shows the ratio of on target editing to off target editing for this guide RNA. The level of on target editing is 71.06-fold greater than the level of off target editing.

[0312] Figure 19B left graph shows percentage editing of the guide RNA targeting the ‘C’ allele of rs2754163 on iPSC lines homozygous for the ‘T’ allele or the ‘C’ allele, the graph on the right shows the ratio of on target editing compared to off target editing for this guide RNA. The level of on target editing is only 4.56-fold greater than the level of off target editing.

[0313] The ratio of on target to off target editing at the rs377373012 4bp indel is therefore substantially higher than the ratio at the rs2754163 SNP.

[0314] Example 8

[0315] Materials and Methods

[0316] Gymnosis in iPS-cardiomyocytes. ASOs were added to the media for 72 hours prior to harvesting for RNA analysis by qPCR.

[0317] Results

[0318] Figure 20 shows the activities of two ASOs in the C series on iPSC-derived cardiomyocytes (Figure 20A shows C1 and Figure 20B shows C4), which were designed to target the MYH7 INDEL rs34598192 (Deletion allele), at doses ranging from 0.5 micromolar to 20 micromolar either on a iPSC derived cardiomyocyte line that is homozygous for the non-target allele (left) or homozygous for the target allele (right). Results show that C1 and C4 effectively and specifically at reduce expression of the target allele. References 1. Martin, M., Ebert, P., & Marschall, T. (2022). Read-Based Phasing and Analysis of Phased Variants with WhatsHap. Methods in Molecular Biology, 2590, 127-138. Humana, New York, NY. https: / / doi.org / 10.1007 / 978-1-0716-2819-5_82. 2. McElwain, M.A., & Peters, B.A. (2023). Accurate Sequencing and Haplotyping from 10 Cells Using Long Fragment Read (LFR) Technology. Laboratory Investigation. https: / / doi.org / 10.1016 / j.labinv.2023.100160 3. Peters, B.A., Liu, J., & Drmanac, R. (2014). Co-barcoded sequence reads from long DNA fragments: a cost-effective solution for "perfect genome" sequencing. Frontiers in Genetics, 5, 466. https: / / doi.org / 10.3389 / fgene.2011.00108 4. Stulz, R., Lerche, M., Luige, O., Taylor, A., Geschwindner, S., & Ghidini, A. (2023). An enhanced biophysical screening strategy to investigate the affinity of ASOs for their target RNA. RSC Chemical Biology, 4(12), 1123-1130.5. Tran, H., Moazami, M. P., Yang, H., McKenna-Yasek, D., Douthwright, C. L., Pinto, C., ... & Brown Jr, R. H. (2022). Suppression of mutant C9orf72 expression by a potent mixed backbone antisense oligonucleotide. Nature medicine, 28(1), 117-124. 6. Anderson, B. R., Jensen, M. L., Hagedorn, P. H., Little, S. C., Olson, R. E., Ammar, R., ... & Bristow, L. J. (2020). Allele-selective knockdown of MYH7 using antisense oligonucleotides. Molecular Therapy Nucleic Acids, 19, 1290-1298. 7. Lian, X., Hsiao, C., Wilson, G., Zhu, K., Hazeltine, L. B., Azarin, S. M., ... & Palecek, S. P. (2012). Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proceedings of the National Academy of Sciences, 109(27), E1848-E1857. 8. Burridge, P. W., Matsa, E., Shukla, P., Lin, Z. C., Churko, J. M., Ebert, A. D., ... & Wu, J. C. (2014). Chemically defined generation of human cardiomyocytes. Nature methods, 11(8), 855-860. 9. Chen, S., Francioli, L. C., Goodrich, J. K., Collins, R. L., Kanai, M., Wang, Q., ... & Karczewski, K. J. (2024). A genomic mutational constraint map using variation in 76,156 human genomes. Nature, 625(7993), 92-100. 10. Brinkman, E. K., Chen, T., Amendola, M., & Van Steensel, B. (2014). Easy quantitative assessment of genome editing by sequence trace decomposition. Nucleic acids research, 42(22), e168-e168.SEQUENCES Phosphonothioate linkage (*) Locked Nucleic Acids, LNA (+) 2'-O-methoxyethyl (MOE) adenine (A), thymine (T), guanine (G), uracil (U) 5-methyl C (C) Antisense Oligonucleotide Sequences Targeting MYH7 SNP rs2239578 SEQ ID 1 (A1): MOE-T*MOE-T*MOE-T*+T*+C*A*A*C*A*C*T*C*T*+A*+G*MOE-C SEQ ID 2 (A2): MOE-T*MOE-T*+T*+T*+C*A*A*C*A*C*T*C*T*+A*+G*MOE-C SEQ ID 3 (A3): MOE-T*MOE-T*+T*+T*+C*A*A*C*A*C*T*C*T*+A*+G*+C SEQ ID 4 (A4): MOE-T*MOE-T*MOE-T*MOE-T*+C*A*A*C*A*C*T*C*T*+A*MOE- G*MOE-C SEQ ID 5 (A5): +T*MOE-T*MOE-T*MOE-T*MOE-C*A*A*C*A*C*T*C*T*MOE-A*MOE- G*+C SEQ ID 6 (A6): +T*+T*+T*T*C*A*A*C*A*C*T*C*T*+A*+G*+C SEQ ID 7 (A7): +A*+T*+T*+T*T*C*A*A*C*A*C*T*C*T*+A*+G*+C*+T SEQ ID 8 (A8): +A*+T*+T*+T*+T*+C*A*A*C*A*C*T*C*+T*+A*+G*+C*+T SEQ ID 9 (A9): MOE-G*MOE-A*MOE-T*MOE-T*MOE- T*T*C*A*A*C*A*C*T*C*T*MOE-A*MOE-G*MOE-C*MOE-T*MOE-T SEQ ID 10 (A10): MOE-G*MOE-A*MOE-T*MOE-T*MOE-T*MOE-T*MOE- C*A*A*C*A*C*T*C*MOE-T*MOE-A*MOE-G*MOE-C*MOE-T*MOE-T SEQ ID 11 (A11): MOE-T*MOE-A*MOE-G*MOE-A*MOE-T*MOE-T*MOE-T*MOE- T*C*A*A*C*A*C*T*C*T*A*MOE-G*MOE-C*MOE-T*MOE-T*MOE-C*MOE-A*MOE-GSEQ ID 12 (A12): MOE-T*MOE-A*MOE-G*MOE-A*MOE-T*MOE-T*MOE-T*MOE- T*MOE-C*A*A*C*A*C*T*C*MOE-T*MOE-A*MOE-G*MOE-C*MOE-T*MOE-T*MOE- C*MOE-A*MOE-G SEQ ID 13 (A13): MOE-A*MOE-C*MOE-T*MOE-A*MOE-G*MOE-A*MOE-T*MOE- T*MOE-T*MOE-T*C*A*A*C*A*C*T*C*T*A*MOE-G*MOE-C*MOE-T*MOE-T*MOE- C*MOE-A*MOE-G*MOE-C*MOE-T*MOE-T SEQ ID 14 (A14): MOE-A*MOE-C*MOE-T*MOE-A*MOE-G*MOE-A*MOE-T*MOE- T*MOE-T*MOE-T*MOE-C*A*A*C*A*C*T*C*MOE-T*MOE-A*MOE-G*MOE-C*MOE- T*MOE-T*MOE-C*MOE-A*MOE-G*MOE-C*MOE-T*MOE-T SEQ ID 15 (A15): MOE-A*MOE-T*MOE-T*MOE-T*T*C*A*A*C*A*C*T*C*T*MOE- A*MOE-G*MOE-C*MOE-T SEQ ID 16 (A16): MOE-A*MOE-T*MOE-T*MOE-T*MOE-T*MOE- C*A*A*C*A*C*T*C*MOE-T*MOE-A*MOE-G*MOE-C*MOE-T SEQ ID 17 (B1): MOE-T*MOE-T*MOE-T*+T*+C*A*A*C*G*C*T*C*T*+A*+G*MOE-C SEQ ID 18 (B2): MOE-T*MOE-T*+T*+T*+C*A*A*C*G*C*T*C*T*+A*+G*MOE-C SEQ ID 19 (B3): MOE-T*MOE-T*+T*+T*+C*A*A*C*G*C*T*C*T*+A*+G*+C SEQ ID 20 (B4): MOE-T*MOE-T*MOE-T*MOE-T*+C*A*A*C*G*C*T*C*T*+A*MOE- G*MOE-C SEQ ID 21 (B5): +T*MOE-T*MOE-T*MOE-T*MOE-C*A*A*C*G*C*T*C*T*MOE-A*MOE- G*+C SEQ ID 22 (B6): +T*+T*+T*T*C*A*A*C*G*C*T*C*T*+A*+G*+C SEQ ID 23 (B7): +A*+T*+T*+T*T*C*A*A*C*G*C*T*C*T*+A*+G*+C*+T SEQ ID 24 (B8): +A*+T*+T*+T*+T*+C*A*A*C*G*C*T*C*+T*+A*+G*+C*+T SEQ ID 25 (B9): MOE-G*MOE-A*MOE-T*MOE-T*MOE- T*T*C*A*A*C*G*C*T*C*T*MOE-A*MOE-G*MOE-C*MOE-T*MOE-TSEQ ID 26 (B10): MOE-G*MOE-A*MOE-T*MOE-T*MOE-T*MOE-T*MOE- C*A*A*C*G*C*T*C*MOE-T*MOE-A*MOE-G*MOE-C*MOE-T*MOE-T SEQ ID 27 (B11): MOE-T*MOE-A*MOE-G*MOE-A*MOE-T*MOE-T*MOE- T*T*C*A*A*C*G*C*T*C*T*MOE-A*MOE-G*MOE-C*MOE-T*MOE-T*MOE-C*MOE- A*MOE-G SEQ ID 28 (B12): MOE-T*MOE-A*MOE-G*MOE-A*MOE-T*MOE-T*MOE-T*MOE- T*MOE-C*A*A*C*G*C*T*C*MOE-T*MOE-A*MOE-G*MOE-C*MOE-T*MOE-T*MOE- C*MOE-A*MOE-G SEQ ID 29 (B13): MOE-T*MOE-T*MOE-A*MOE-C*MOE-T*MOE-A*MOE-G*MOE- A*MOE-T*MOE-T*MOE-T*T*C*A*A*C*G*C*T*C*T*MOE-A*MOE-G*MOE-C*MOE- T*MOE-T*MOE-C*MOE-A*MOE-G*MOE-C SEQ ID 30 (B14): MOE-T*MOE-T*MOE-A*MOE-C*MOE-T*MOE-A*MOE-G*MOE- A*MOE-T*MOE-T*MOE-T*MOE-T*MOE-C*A*A*C*G*C*T*C*MOE-T*MOE-A*MOE- G*MOE-C*MOE-T*MOE-T*MOE-C*MOE-A*MOE-G*MOE-C SEQ ID 31 (B15): MOE-A*MOE-T*MOE-T*MOE-T*T*C*A*A*C*G*C*T*C*T*MOE- A*MOE-G*MOE-C*MOE-T SEQ ID 32 (B16): MOE-A*MOE-T*MOE-T*MOE-T*MOE-T*MOE- C*A*A*C*G*C*T*C*MOE-T*MOE-A*MOE-G*MOE-C*MOE-T Antisense Oligonucleotide Sequences Targeting MYH7 indel rs34598192 SEQ ID 33 (C1): +A*+A*+G*G*G*A*A*G*T*G*A*T*T*+T*+G*+A SEQ ID 34 (C2): +A*+G*+G*G*A*A*G*T*G*A*T*T*T*+G*+A*+T SEQ ID 35 (C3): +G*+G*+A*A*G*T*G*A*T*T*T*G*A*+T*+G*+C SEQ ID 36 (C4): +G*+A*+T*T*T*G*A*T*G*C*A*A*G*+G*+C*+T SEQ ID 37 (C5): +A*+T*T*T*G*A*T*G*C*A*A*G*G*+C*+T*A SEQ ID 38 (C6): +A*+G*+G*+G*A*+A*+G*T*G*A*T*T*T*G*+A*+TSEQ ID 39 (C7): +A*+G*+G*G*A*+A*+G*T*G*A*T*T*T*G*+A*+T SEQ ID 40 (C8): +G*+A*T*T*T*G*A*T*G*+C*+A*+A*+G*+G*+C*+T SEQ ID 41 (C9): +G*+A*T*T*T*G*A*T*G*+C*+A*A*G*+G*+C*+T SEQ ID 42 (C10): +A*+G*+G*+G*+A*A*G*T*G*A*T*T*T*+G*+A*+T*+G*+C SEQ ID 43 (C11): +A*+A*+G*+T*+G*A*T*T*T*G*A*T*G*+C*+A*+A*+G*+G SEQ ID 44 (C12): +A*+G*+G*G*+A*+A*G*T*G*A*T*T*T*+G*+A*+T*+G*+C SEQ ID 45 (C13): +A*+A*+G*T*G*A*T*T*T*G*+A*+T*+G*+C*+A*+A*+G*+G SEQ ID 46 (C14): +G*+A*+A*+G*+G*+G*A*A*G*T*G*A*T*T*T*G*+A*+T SEQ ID 47 (C15): +G*+A*+A*+G*+G*+G*+A*+A*+G*T*G*A*T*T*T*G*A*T SEQ ID 48 (C16): +G*+T*G*A*T*T*T*G*A*T*G*C*+A*+A*+G*+G*+C*T SEQ ID 49 (C17): +G*+T*G*A*T*T*T*G*A*+T*+G*+C*+A*+A*+G*+G*+C*+T SEQ ID 50 (C18): MOE-A*MOE-G*MOE-G*MOE-G*MOE- A*A*G*T*G*A*T*T*T*G*A*MOE-T*MOE-G*MOE-C*MOE-A*MOE-A SEQ ID 51 (C19): MOE-A*MOE-G*MOE-G*MOE-G*MOE-A*MOE-A*MOE-G*MOE- T*G*A*T*T*T*G*A*MOE-T*MOE-G*MOE-C*MOE-A*MOE-A SEQ ID 52 (C20): MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A*MOE- A*G*T*G*A*T*T*T*G*A*T*MOE-G*MOE-C*MOE-A*MOE-A*MOE-G*MOE-G*MOE- C*MOE-T SEQ ID 53 (C21): MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A*MOE-A*MOE- G*MOE-T*G*A*T*T*T*G*A*MOE-T*MOE-G*MOE-C*MOE-A*MOE-A*MOE-G*MOE- G*MOE-C*MOE-T SEQ ID 54 (C22): MOE-G*MOE-G*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE- A*MOE-A*MOE-G*T*G*A*T*T*T*G*A*T*G*MOE-C*MOE-A*MOE-A*MOE-G*MOE- G*MOE-C*MOE-T*MOE-A*MOE-G*MOE-TSEQ ID 55 (C23): MOE-G*MOE-G*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A*MOE-A*MOE- G*MOE-T*G*A*T*T*T*G*A*MOE-T*MOE-G*MOE-C*MOE-A*MOE-A*MOE-G*MOE- G*MOE-C*MOE-T*MOE-A*MOE-G*MOE-T SEQ ID 56 (C24): MOE-G*MOE-A*MOE-A*MOE-G*MOE-G*MOE- G*A*A*G*T*G*A*T*T*T*G*MOE-A*MOE-T SEQ ID 57 (C25): MOE-G*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A*MOE- A*MOE-G*T*G*A*T*T*T*G*MOE-A*MOE-T SEQ ID 58 (C26): MOE-G*MOE-T*G*A*T*T*T*G*A*T*G*C*MOE-A*MOE-A*MOE- G*MOE-G*MOE-C*MOE-T SEQ ID 59 (C27): MOE-G*MOE-T*G*A*T*T*T*G*A*MOE-T*MOE-G*MOE-C*MOE- A*MOE-A*MOE-G*MOE-G*MOE-C*MOE-T SEQ ID 60 (D1): +A*+A*+G*G*G*A*A*G*T*G*A*T*+G*+T*+T SEQ ID 61 (D2): +A*+G*+G*G*A*A*G*T*G*A*T*G*T*+T*+G*+A SEQ ID 62 (D3): +G*+G*+G*A*A*G*T*G*A*T*G*T*T*+G*+A*+T SEQ ID 63 (D4): +T*+G*+A*T*G*T*T*G*A*T*G*C*A*+A*+G*+G SEQ ID 64 (D5): +G*+A*+T*G*T*T*G*A*T*G*C*A*A*+G*+G*+C SEQ ID 65 (D6): +A*+G*G*+G*+A*A*+G*T*G*A*T*G*T*+T*+G*+A SEQ ID 66 (D7): +A*+G*+G*+G*+A*+A*G*T*G*A*T*G*T*+T*+G*+A SEQ ID 67 (D8): +A*+G*MOE-G*MOE-G*MOE-A*MOE-A*G*T*G*A*T*G*T*MOE- T*MOE-G*+A SEQ ID 68 (D9): MOE-A*MOE-G*MOE-G*MOE-G*MOE-A*MOE- A*G*T*G*A*T*G*T*MOE-T*MOE-G*MOE-A SEQ ID 69 (D10): +G*+A*+A*+G*T*G*A*T*G*T*T*G*A*T*+G*+C*+A*+ASEQ ID 70 (D11): +G*+A*+A*+G*+T*G*A*T*G*T*T*G*+A*+T*+G*+C*+A*+A SEQ ID 71 (D12): MOE-G*MOE-G*MOE-A*MOE-A*MOE- G*T*G*A*T*G*T*T*G*A*T*MOE-G*MOE-C*MOE-A*MOE-A*MOE-G SEQ ID 72 (D13): MOE-G*MOE-G*MOE-A*MOE-A*MOE-G*MOE- T*G*A*T*G*T*T*G*MOE-A*MOE-T*MOE-G*MOE-C*MOE-A*MOE-A*MOE-G SEQ ID 73 (D14): MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A*MOE-A*MOE- G*T*G*A*T*G*T*T*G*A*T*MOE-G*MOE-C*MOE-A*MOE-A*MOE-G*MOE-G*MOE-C SEQ ID 74 (D15): MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A*MOE-A*MOE- G*MOE-T*G*A*T*G*T*T*G*MOE-A*MOE-T*MOE-G*MOE-C*MOE-A*MOE-A*MOE- G*MOE-G*MOE-C SEQ ID 75 (D16): MOE-G*MOE-G*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE- A*MOE-A*MOE-G*T*G*A*T*G*T*T*G*A*T*MOE-G*MOE-C*MOE-A*MOE-A*MOE- G*MOE-G*MOE-C*MOE-T*MOE-A*MOE-G SEQ ID 76 (D17): MOE-G*MOE-G*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE- A*MOE-A*MOE-G*MOE-T*G*A*T*G*T*T*G*MOE-A*MOE-T*MOE-G*MOE-C*MOE- A*MOE-A*MOE-G*MOE-G*MOE-C*MOE-T*MOE-A*MOE-G SEQ ID 77 (D18): MOE-G*MOE-A*MOE-A*MOE-G*T*G*A*T*G*T*T*G*A*T*MOE- G*MOE-C*MOE-A*MOE-A SEQ ID 78 (D19): MOE-G*MOE-A*MOE-A*MOE-G*MOE-T*G*A*T*G*T*T*G*MOE- A*MOE-T*MOE-G*MOE-C*MOE-A*MOE-A Editing Sequences SEQ ID 79 gRNA: AAAUCACUUCCCUUCCCAGCGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUUU SEQ ID 80 gRNA:ACAUCACUUCCCUUCCCAGCGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUUU shRNA Sequences SEQ ID 81 shRNA: UCAAAUCACUUCCCUUCCCAGUUCAAGAGACUGGGAAGGAGUGAUUUGAA SEQ ID 82 shRNA: CAACAUCACUUCCCUUCCCAGUUCAAGAGACUGGGAAGGAGUGAUGUUGG SEQ ID 83 shRNA: UCAAACUUCCCUUCCCAGCAUUCAAGAGAUGCUGGGAAGGAAAGUUGAA SEQ ID 84 shRNA: CAACAUUCCCUUCCCAGCAAUUCAAGAGAUUUGCUGGGAAGGAAUGUUGG Antisense Oligonucleotide Sequences Targeting TNNI3 Indel rs377373012 SEQ ID 85 (E1): +G*+T*MOE-G*MOE-T*MOE-C*C*T*G*T*C*T*G*T*C*T*MOE- C*MOE-C*MOE-T*+A*+A SEQ ID 86 (E2): +T*+G*MOE-T*MOE-C*MOE-C*T*G*T*C*T*G*T*C*T*C*MOE- C*MOE-T*MOE-A*+A*+G SEQ ID 87 (E3): +G*+T*MOE-C*MOE-C*MOE-T*G*T*C*T*G*T*C*T*C*C*MOE- T*MOE-A*MOE-A*+G*+G SEQ ID 88 (E4): +T*+C*MOE-C*MOE-T*MOE-G*T*C*T*G*T*C*T*C*C*T*MOE- A*MOE-A*MOE-G*+G*+G SEQ ID 89 (E5): +C*+C*MOE-T*MOE-G*MOE-T*C*T*G*T*C*T*C*C*T*A*MOE- A*MOE-G*MOE-G*+G*+A SEQ ID 90 (E6): +C*+T*MOE-G*MOE-T*MOE-C*T*G*T*C*T*C*C*T*A*A*MOE- G*MOE-G*MOE-G*+A*+CSEQ ID 91 (E7): +T*+G*MOE-T*MOE-C*MOE-T*G*T*C*T*C*C*T*A*A*G*MOE- G*MOE-G*MOE-A*+C*+C SEQ ID 92 (E8): +G*+T*MOE-C*C*T*G*T*C*T*G*T*C*T*MOE-C*+C*+T SEQ ID 93 (E9): +T*+C*MOE-C*T*G*T*C*T*G*T*C*T*C*MOE-C*+T*+A SEQ ID 94 (E10): +C*+C*MOE-T*G*T*C*T*G*T*C*T*C*C*MOE-T*+A*+A SEQ ID 95 (E11): +C*+T*MOE-G*T*C*T*G*T*C*T*C*C*T*MOE-A*+A*+G SEQ ID 96 (E12): +T*+G*MOE-T*C*T*G*T*C*T*C*C*T*A*MOE-A*+G*+G SEQ ID 97 (E13): +G*+T*MOE-C*T*G*T*C*T*C*C*T*A*A*MOE-G*+G*+G SEQ ID 98 (E14): +T*+C*MOE-T*G*T*C*T*C*C*T*A*A*G*MOE-G*+G*+A SEQ ID 99 (E15): +C*+C*+T*G*T*C*T*G*T*C*T*C*C*+T*+A*+A SEQ ID 100 (E16): +C*+T*+G*+T*+C*T*G*T*C*T*C*C*+T*+A*+A*+G*+G*+G SEQ ID 101 (E17): MOE-C*MOE-T*MOE-G*MOE-T*MOE-C*T*G*T*C*T*C*C*MOE- T*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G SEQ ID 102 (E18): +C*+T*+G*T*C*T*G*T*C*T*+C*+C*+T*+A*+A*+G*+G*+G SEQ ID 103 (E19): MOE-C*MOE-T*MOE-G*T*C*T*G*T*C*T*MOE-C*MOE-C*MOE- T*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G SEQ ID 104 (E20): +C*+T*G*U*C*T*G*+T*+C*T*C*+C*+T*A*A*+G*+G*+G SEQ ID 105 (E21): MOE-C*MOE-T*G*U*C*T*G*MOE-T*MOE-C*T*C*MOE-C*MOE- T*A*A*MOE-G*MOE-G*MOE-G SEQ ID 106 (E22 / ASO16): MOE-G*MOE-C*MOE-T*MOE-G*MOE-T*MOE-G*MOE- T*MOE-C*MOE-C*T*G*T*C*T*G*T*C*T*C*MOE-C*MOE-T*MOE-A*MOE-A*MOE- G*MOE-GSEQ ID 107 (E23): MOE-G*MOE-G*MOE-G*MOE-C*MOE-T*MOE-G*MOE-T*MOE- G*T*C*C*T*G*T*C*T*G*T*MOE-C*MOE-T*MOE-C*MOE-C*MOE-T*MOE-A*MOE- A*MOE-G*MOE-G*MOE-G*MOE-A*MOE-C SEQ ID 108 (H1): +T*+C*+C*T*+G*T*C*T*C*C*T*A*A*+G*+G*+G SEQ ID 109 (H2): +T*+C*+C*+T*+G*+T*C*T*C*C*T*A*A*+G*+G*+G SEQ ID 110 (H3): +T*+C*+C*+T*G*T*C*T*C*C*T*A*A*G*+G*+G*+A*+C SEQ ID 111 (H4): +T*+C*+C*+T*+G*+T*C*T*C*C*T*A*A*G*G*G*+A*+C SEQ ID 112 (H5): +T*+C*C*T*G*T*C*T*C*C*T*A*+A*+G*+G*+G*+A*+C SEQ ID 113 (H6): +T*+C*+C*+T*+G*+T*C*T*C*C*T*A*A*+G*+G*+G*+A*+C SEQ ID 114 (H7): +T*+C*+C*+T*+G*+T*+C*+T*C*C*T*A*A*G*G*+G*+A*+C SEQ ID 115 (H8): +T*+C*+C*T*G*T*C*T*C*C*+T*+A*+A*+G*+G*+G*+A*+C SEQ ID 116 (H9): MOE-T*MOE-C*MOE-C*MOE-T*G*T*C*T*C*C*T*A*A*G*MOE- G*MOE-G*MOE-A*MOE-C SEQ ID 117 (H10): MOE-T*MOE-C*MOE-C*MOE-T*MOE-G*MOE- T*C*T*C*C*T*A*A*G*G*G*MOE-A*MOE-C SEQ ID 118 (H11): MOE-T*MOE-C*C*T*G*T*C*T*C*C*T*A*MOE-A*MOE-G*MOE- G*MOE-G*MOE-A*MOE-C SEQ ID 119 (H12): MOE-T*MOE-C*MOE-C*MOE-T*MOE-G*MOE- T*C*T*C*C*T*A*A*MOE-G*MOE-G*MOE-G*MOE-A*MOE-C SEQ ID 120 (H13): MOE-T*MOE-C*MOE-C*MOE-T*MOE-G*MOE-T*MOE-C*MOE- T*C*C*T*A*A*G*G*MOE-G*MOE-A*MOE-C SEQ ID 121 (H14): MOE-T*MOE-C*MOE-C*T*G*T*C*T*C*C*MOE-T*MOE-A*MOE- A*MOE-G*MOE-G*MOE-G*MOE-A*MOE-CSEQ ID 122 (H15): MOE-C*MOE-T*MOE-G*MOE-T*MOE- G*T*C*C*T*G*T*C*T*C*C*MOE-T*MOE-A*MOE-A*MOE-G*MOE-G SEQ ID 123 (H16): MOE-C*MOE-T*MOE-G*MOE-T*MOE-G*MOE-T*MOE-C*MOE- C*T*G*T*C*T*C*C*T*A*A*MOE-G*MOE-G SEQ ID 124 (H17): MOE-C*MOE-T*G*T*G*T*C*C*T*G*T*C*MOE-T*MOE-C*MOE- C*MOE-T*MOE-A*MOE-A*MOE-G*MOE-G SEQ ID 125 (H18): MOE-C*MOE-T*MOE-G*MOE-T*MOE-G*MOE- T*C*C*T*G*T*C*T*C*MOE-C*MOE-T*MOE-A*MOE-A*MOE-G*MOE-G SEQ ID 126 (H19): MOE-C*MOE-T*MOE-G*T*G*T*C*C*T*G*MOE-T*MOE-C*MOE- T*MOE-C*MOE-C*MOE-T*MOE-A*MOE-A*MOE-G*MOE-G SEQ ID 127 (H20): MOE-C*MOE-T*MOE-G*MOE-T*MOE-G*MOE-T*MOE-C*MOE- C*MOE-T*MOE-G*T*C*T*C*C*T*A*MOE-A*MOE-G*MOE-G SEQ ID 128 (H21): MOE-T*MOE-G*MOE-T*MOE-G*MOE- T*C*C*T*G*T*C*T*C*C*T*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G SEQ ID 129 (H22): MOE-T*MOE-G*T*G*T*C*C*T*G*T*C*T*MOE-C*MOE-C*MOE- T*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G SEQ ID 130 (H23): MOE-T*MOE-G*MOE-T*MOE-G*MOE-T*MOE-C*MOE-C*MOE- T*G*T*C*T*C*C*T*A*A*G*MOE-G*MOE-G SEQ ID 131 (H24): MOE-T*MOE-G*MOE-T*MOE-G*MOE-T*MOE-C*MOE- C*T*G*T*C*T*C*C*MOE-T*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G SEQ ID 132 (H25): MOE-T*MOE-G*MOE-T*G*T*C*C*T*G*T*MOE-C*MOE-T*MOE- C*MOE-C*MOE-T*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G SEQ ID 133 (H26): MOE-T*MOE-G*MOE-T*MOE-G*MOE-T*MOE-C*MOE-C*MOE- T*MOE-G*MOE-T*C*T*C*C*T*A*A*MOE-G*MOE-G*MOE-G SEQ ID 134 (H27): MOE-G*MOE-T*MOE-G*MOE-T*MOE- C*C*T*G*T*C*T*C*C*T*A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-ASEQ ID 135 (H28): MOE-G*MOE-T*G*T*C*C*T*G*T*C*T*C*MOE-C*MOE-T*MOE- A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A SEQ ID 136 (H29): MOE-G*MOE-T*MOE-G*MOE-T*MOE-C*MOE-C*MOE-T*MOE- G*T*C*T*C*C*T*A*A*G*G*MOE-G*MOE-A SEQ ID 137 (H30): MOE-G*MOE-T*MOE-G*MOE-T*MOE-C*MOE-C*MOE- T*G*T*C*T*C*C*T*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A SEQ ID 138 (H31): MOE-G*MOE-T*MOE-G*T*C*C*T*G*T*C*MOE-T*MOE-C*MOE- C*MOE-T*MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*MOE-A SEQ ID 139 (H32): MOE-G*MOE-T*MOE-G*MOE-T*MOE-C*MOE-C*MOE-T*MOE- G*MOE-T*MOE-C*T*C*C*T*A*A*G*MOE-G*MOE-G*MOE-A Editing sequences SEQ ID 140 gRNA: AGGACACAGCCCACCACUAAGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUUU SEQ ID 141 gRNA: ACACAGCCCACCACUAACCCGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUUU shRNAs SEQ ID 142 shRNA: UCAAAUCACUUCCCUUCCCAGUUCAAGAGACUGGGAAGGAGUGAUUUGAA SEQ ID 143 shRNA: CAACAUCACUUCCCUUCCCAGUUCAAGAGACUGGGAAGGAGUGAUGUUGG SEQ ID 144 shRNA: UCAAACUUCCCUUCCCAGCAUUCAAGAGAUGCUGGGAAGGAAAGUUGAASEQ ID 145 shRNA: CAACAUUCCCUUCCCAGCAAUUCAAGAGAUUUGCUGGGAAGGAAUGUUGG Antisense Oligonucleotide Sequences Targeting TNNT2 Indel rs45533739 SEQ ID 146 (F1): +T*+C*+T*G*C*T*C*A*G*A*A*G*A*+G*+A*+A SEQ ID 147 (F2): +T*+C*+A*G*A*A*G*A*G*A*A*G*T*+C*+C*+A SEQ ID 148 (F3): +C*+A*+G*A*A*G*A*G*A*A*G*T*C*+C*+A*+G SEQ ID 149 (F4): +C*+T*+C*+A*G*A*A*G*A*G*A*A*G*T*+C*+C*+A*+G SEQ ID 150 (F5): +C*+T*C*A*G*A*A*G*A*G*A*A*+G*+T*+C*+C*+A*+G SEQ ID 151 (F6): +C*+T*+C*+A*+G*A*A*G*A*G*A*A*+G*+T*+C*+C*+A*+G SEQ ID 152 (F7): +C*+T*+C*A*G*A*A*G*A*G*+A*+A*+G*+T*+C*+C*+A*+G SEQ ID 232 (F8): MOE-C*MOE-T*MOE-C*MOE-A*MOE- G*A*A*G*A*G*A*A*G*T*C*MOE-C*MOE-A*MOE-G*MOE-G*MOE-C SEQ ID 153 (F9): MOE-C*MOE-T*C*A*G*A*A*G*A*G*A*A*MOE-G*MOE-T*MOE- C*MOE-C*MOE-A*MOE-G*MOE-G*MOE-C SEQ ID 154 (F10): MOE-C*MOE-T*MOE-C*MOE-A*MOE-G*A*A*G*A*G*A*A*MOE- G*MOE-T*MOE-C*MOE-C*MOE-A*MOE-G*MOE-G*MOE-C SEQ ID 155 (F11): MOE-C*MOE-T*MOE-C*A*G*A*A*G*A*G*MOE-A*MOE-A*MOE- G*MOE-T*MOE-C*MOE-C*MOE-A*MOE-G*MOE-G*MOE-C SEQ ID 156 (F12): MOE-T*MOE-T*MOE-C*MOE-T*MOE-G*MOE-C*MOE-T*MOE- C*A*G*A*A*G*A*G*A*A*G*MOE-T*MOE-C*MOE-C*MOE-A*MOE-G*MOE-G*MOE-C SEQ ID 157 (F13): MOE-T*MOE-T*MOE-C*MOE-T*MOE-G*MOE-C*MOE-T*MOE- C*MOE-A*G*A*A*G*A*G*A*MOE-A*MOE-G*MOE-T*MOE-C*MOE-C*MOE-A*MOE- G*MOE-G*MOE-CSEQ ID 158 (F14): MOE-C*MOE-T*MOE-C*MOE-A*G*A*A*G*A*G*A*A*G*T*MOE- C*MOE-C*MOE-A*MOE-G SEQ ID 159 (F15): MOE-C*MOE-T*C*A*G*A*A*G*A*G*A*A*MOE-G*MOE-T*MOE- C*MOE-C*MOE-A*MOE-G SEQ ID 160 (F16): MOE-C*MOE-T*MOE-C*MOE-A*MOE-G*A*A*G*A*G*A*A*MOE- G*MOE-T*MOE-C*MOE-C*MOE-A*MOE-G SEQ ID 161 (F17): MOE-C*MOE-T*MOE-C*A*G*A*A*G*A*G*MOE-A*MOE-A*MOE- G*MOE-T*MOE-C*MOE-C*MOE-A*MOE-G SEQ ID 162 (G1): +C*+T*+G*C*T*C*A*G*A*A*G*T*C*+C*+A*+G SEQ ID 163 (G2): +C*+T*+G*+C*T*C*A*G*A*A*G*T*C*C*+A*+G SEQ ID 164 (G3): +C*+T*+G*+C*+T*+C*+A*G*A*A*G*T*C*C*+A*+G SEQ ID 165 (G4): +T*+G*+C*T*C*A*G*A*A*G*T*C*C*+A*+G*+G SEQ ID 166 (G5): +T*+G*C*T*C*A*G*A*A*+G*+T*+C*+C*+A*+G*+G SEQ ID 167 (G6): +T*+G*+C*+T*C*A*G*A*A*G*T*C*C*A*+G*+G*+C*+A SEQ ID 168 (G7): +T*+G*C*T*C*A*G*A*A*G*T*C*+C*+A*+G*+G*+C*+A SEQ ID 169 (G8): +T*+G*C*T*C*A*G*A*A*+G*+T*+C+*C+*A+*G+*G+*C+*A SEQ ID 170 (G9): MOE-C*MOE-T*G*C*T*C*A*G*A*A*G*T*MOE-C*MOE-C*MOE- A*MOE-G*MOE-G*MOE-C*MOE-A*MOE-G SEQ ID 171 (G10): MOE-C*MOE-T*G*C*T*C*A*G*A*MOE-A*MOE-G*MOE-T*MOE- C*MOE-C*MOE-A*MOE-G*MOE-G*MOE-C*MOE-A*MOE-G SEQ ID 172 (G11): MOE-G*MOE-C*MOE-T*MOE-T*MOE-C*MOE-T*MOE- G*C*T*C*A*G*A*A*G*T*C*MOE-C*MOE-A*MOE-G*MOE-G*MOE-C*MOE-A*MOE- G*MOE-CSEQ ID 173 (G12): MOE-G*MOE-C*MOE-T*MOE-T*MOE-C*MOE-T*MOE- G*C*T*C*A*G*A*A*MOE-G*MOE-T*MOE-C*MOE-C*MOE-A*MOE-G*MOE-G*MOE- C*MOE-A*MOE-G*MOE-C SEQ ID 174 (G13): MOE-C*MOE-T*MOE-G*MOE-C*MOE-T*MOE-T*MOE-C*MOE- T*MOE-G*MOE-C*T*C*A*G*A*A*G*T*C*C*MOE-A*MOE-G*MOE-G*MOE-C*MOE- A*MOE-G*MOE-C*MOE-A*MOE-A*MOE-G SEQ ID 175 (G14): MOE-C*MOE-T*MOE-G*MOE-C*MOE-T*MOE-T*MOE-C*MOE- T*MOE-G*MOE-C*MOE-T*C*A*G*A*A*G*T*MOE-C*MOE-C*MOE-A*MOE-G*MOE- G*MOE-C*MOE-A*MOE-G*MOE-C*MOE-A*MOE-A*MOE-G SEQ ID 176 (G15): MOE-T*MOE-G*MOE-C*MOE-T*C*A*G*A*A*G*T*C*C*A*MOE- G*MOE-G*MOE-C*MOE-A SEQ ID 177 (G16): MOE-T*MOE-G*MOE-C*MOE-T*MOE-C*A*G*A*A*G*T*C*MOE- C*MOE-A*MOE-G*MOE-G*MOE-C*MOE-A SEQ ID 178 (G17): MOE-T*MOE-G*C*T*C*A*G*A*A*G*T*C*MOE-C*MOE-A*MOE- G*MOE-G*MOE-C*MOE-A SEQ ID 179 (G18): MOE-T*MOE-G*C*T*C*A*G*A*A*MOE-G*MOE-T*MOE-C*MOE- C*MOE-A*MOE-G*MOE-G*MOE-C*MOE-A SEQ ID 180 (CTL16): +C*+G*+C*T*A*C*G*A*T*C*G*A*C*+G*+A*+T SEQ ID 181 (CTL20):+C*+G*+C*+T*+A*C*G*A*T*C*G*A*C*G*A*+T*+G*+T*+T*+G Editing sequences SEQ ID 182 gRNA: CUCUUCUCUUCUGAGCAGAGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUUU SEQ ID 183 gRNA:CUGAGCAGAAGCAGCUGUUGGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGG CUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUUU shRNAs SEQ ID 184 shRNA: CUUCUCUUCUCUUGCUGCCUGUUCAAGAGACAGGCAGCAAGAGAAGAGGA SEQ ID 185 shRNA: CUGAGCAGAAGCAGCUGUUGAUUCAAGAGAUCAACAGCUGCUUCUGCUGA SEQ ID 186 shRNA: UCUUCUCUUGCUGCCUGGACUUUCAAGAGAAGUCCAGGCAGCAAGAGAAGA SEQ ID 187 shRNA: GAGCAGAAGCAGCUGUUGAAGUUCAAGAGACUCAACAGCUGCUUCUGCUGA Antisense Oligonucleotide Sequences Targeting HTT Indel rs59464879 SEQ ID 188: +A*+A*+A*G*G*G*A*C*T*G*G*C*A*+G*+C*+C SEQ ID 189: MOE-A*MOE-A*MOE-A*MOE-G*MOE-G*G*A*C*T*G*G*C*A*G*C*MOE- C*MOE-A*MOE-G*MOE-C*MOE-A SEQ ID 190: +A*+A*+G*G*G*A*A*A*C*T*C*C*T*G*G*C*A*+G*+C*+C SEQ ID 191: MOE-A*MOE-A*MOE-G*MOE-G*MOE-G*A*A*A*C*T*C*C*T*G*G*MOE- C*MOE-A*MOE-G*MOE-C*MOE-C Editing sequences SEQ ID 192 gRNA: CCAGCAUCUGGGGGUGGGAAGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGG CUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUU SEQ ID 193 gRNA:CUGGCAGCCAGCAUCUGGGGGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGG CUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUU SEQ ID 194 gRNA: GUGGAAAGGGAAACUCCUGGGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGG CUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUU shRNAs SEQ ID 195 shRNA: GGCUUGCCAGUCCCUUUUCAAGAGAAAAGGGACUGGCAAGCC SEQ ID 196 shRNA: UGCUUGCCAGUCCCUUUUCAAGAGAAAAGGGACUGGCAAGCCAGCA SEQ ID 197 shRNA: GGCUUGCCAGAGUUUCCUUCAAGAGAAAGGGAAACUCCUGGCAAGCC SEQ ID 198 shRNA: UGCUUGCCAGAGUUUCCUUCAAGAGAAAGGGAAACUCCUGGCAAAGCC Antisense Oligonucleotide Sequences Targeting HTT Indel rs751205475 SEQ ID 199: +C*+A*+T*A*C*A*A*C*G*A*G*C*A*+C*+C*+T SEQ ID 200: MOE-C*MOE-A*MOE-T*MOE-A*MOE-C*A*A*C*G*A*G*C*A*C*C*MOE- T*MOE-C*MOE-C*MOE-A*MOE-A SEQ ID 201: +C*+A*+T*A*C*A*A*G*A*G*C*A*C*+C*+T*+C SEQ ID 202: MOE-C*MOE-A*MOE-T*MOE-A*MOE-C*A*A*G*A*G*C*A*C*C*T*MOE- C*MOE-C*MOE-A*MOE-A*MOE-A Editing sequences SEQ ID 203 gRNA: UGGUUUGGAGGUGCUCUUGUGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGG CUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUU SEQ ID 204 gRNA:GGUGCUCUUGUAUGGUUUGGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUU SEQ ID 205 gRNA: UGGAGGUGCUCUUGUAUGGGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUU SEQ ID 206 gRNA: UGCAGGUGCUCUAUUGCAGGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUU SEQ ID 207 gRNA: UGGAGGUGCUCUGUUGUAGGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGC UAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUUUUUU shRNAs SEQ ID 208: AGGUGCUCGUUGUAUGUUCAAGAGACAUACAACGAGCACC SEQ ID 209: UUGGAGAGGUGCUCGUAUGUUCAAGAGACAUACGAGCACCUCUCCAA SEQ ID 210: AGGAGGUGCUCUUGUAUGUUCAAGAGACAUACAAGAGCACC SEQ ID 211: GUGGAGGUGCUCUGUAUGUUCAAGAGACAUACAGAGCACCUCACC Antisense Oligonucleotide Sequences Targeting HTT Indel rs78373442 SEQ ID 212: +T*+T*+T*G*T*A*T*T*C*A*T*A*A*+A*+A*+G SEQ ID 213: MOE-T*MOE-T*MOE-T*MOE-G*MOE-T*MOE- A*T*T*C*A*T*A*A*A*A*MOE-G*MOE-A*MOE-G*MOE-A SEQ ID 214:+T*+T*+G*T*A*T*T*A*A*T*A*T*T*+C*+A*+T SEQ ID 215: MOE-T*MOE-T*MOE-G*MOE-T*MOE-A*T*T*A*A*T*A*T*T*C*A*MOE- T*MOE-A*MOE-A*MOE-A*MOE-AshRNA SEQ ID 216: CUUUUAUGAAUUACAAAUUCAAGAGAUUUGUAAUUCAUAAAAG SEQ ID 217: UUCUUCUAUGAAUACAAAUUCAAGAGAUUUGUAUUCAUAAGAAGAA SEQ ID 218: AUUGAUUAUAAUUACAAUUCAAGAGAUUUGUAAUUAUAAUUCAU SEQ ID 219: AUUCUUUAUAUAAUUACAUUCAAGAGAUUUGUAAUUAUAUUCAUAA Wild Type TNNT2 gene sequence with bold exons SEQ ID 220: TNNT2 ENSG00000118194 >chromosome:GRCh38:1:201359008:201377764:-1 – indels are indicated by square brackets GCACATTCCTCCCTCCGCAGGGCTGGCTCACCAGGCCCCAGCCCACATGCCTGCT TAAAGCCCTCTCCATCCTCTGCCTCACCCAGTCCCCGCTGAGACTGAGCAGACGCC TCCAGGATCTGTCGGCAGCTGCTGTTCTGAGGTAAGGCTCGGGCAGGGCTCTGGGGA AGAGGAGAGCAGAGAATGGACGGGGAGATGTGAGGGTCTTGGGCCCTGGCATATTTACC CAGAGTCTGCCTGTGTCCGCAGAAGTCCATGGCCCCTCCTGGTGGAGGCCACACTTCAG AGGACAGGTTGCCAGGTCTGGGCTCCAAGATTGGTACAATAGAGCAGAGAGAAAGAGA GAATCCCATGGTAGCCATTGGAGACTAGGGTTGGAGTCCTGGACCTGCCACTAAAGATG TAGAGTTTAAACTCTTGGATGAGTCACTTTCCCTCCCCAGGCCTCAGTTTTCTCATCTGTA GAGTGGGCTAAATATTTGCTAGGCCTCTGCCTGCTCTAAATACAATGAGGCTCTAACCAA CAGAGACTCCCCTAAATTGCTTGTTCCAGGCCGAGGGTCACCTATACCTTTCCAGTCA GGTCCTTCTTGAGTCCCAGGGAGCAGAGCGAGACTGCAGCCCTGTCAGTCAAGTG AGTCTCCTGGTCATATTCTCAAGGCTAGGAACTCAGAAACTGTCCTGGAAGTACTGAGA ACGACCAGATGTGCAGGGTCCTCTGCAGCGCCTCTCCAGGCCAACGCGGGAATTCCATT TGATGCCGCTGACTCAGTGCCATTCCCTCCCAGAGCTGAGCTGAGCTCCCAGACCCTGA GCTGAGCTGCACTTGGGGGGCAGATGGGGAAAGGCCCCACTGGGAGAACCTGTCCTTG GGTGTGAGCCCAGCTGGTTGGCATTAAAGGGTTTAACCTGGGCCCCCGTAAGTGTAACC CTTGTCCCATCCCACCCCAACACCCCATCTTGGGACAAGCCTTGCTCTGTGCCCAGGAGT GGTAGAGAAAGATTCCCTTGGTTTCTAGTTTCTCCTGGGCAGGCTGAGGACTATTCAGGG AGGGGGTTGCCTGGTGTGTAAGGAATTTCAGAGCACATTCCTGGCCTGCTGTAGCCGGG AAGCTACCTGCCCTCCCAAATTCCTTTTCTGACCCATGTCTTAGAGGGCGAATCTTCCTTC TCTCTGCAAAAGTGGAATCTGAGAGGATCAGGTTGCTGGCTGCCTCCTATTTCTGACCCC CACTTCCACTCTTCCTGCCCTCTCCCCATCCCCTTACCCCCGCTCTCCTGGTGGGCTAACAGCTGTTGGCTTAGGAAAATCCCATTCGAGTGCTGGGGACAAGGACTGAGGGGTGGGGCC TGTGTTTCTCAGAAACTCGGAGACTGTTTCTACAGGGACAGAACTCAAGGCTCTCCTGC TTTGTAGTTGGTTAACTGAAGTCAGAGAGGCTAGGTGACTTAAATAAGGCCACACAGCA AAATCTGGGTTTTAAGCCCAGATTTTTTGACCCCAAGGTCAGTATGCCTTATGCCTTCTAT TGTCTTTGCTTTACTCCCCAAGAAAGGTGACCCTGTGAACGTGTTAGGCTTCCAACACGT GTTGGCTATGAATGCATTAGGCTAGAAAGCTGCCTGTGGAAGTGGGTGGGACTTGGGTG TGAGGACACAGGTGTTTCAGGACATTCTGGTTCCTTGCCCTGCCAGGTTCTTGACTCTCT CCCATTCTGCAGCCCCCAGTCTCAGCCCACCTCTTTCCCCTGCATCAGGTCCTGCCTTTG AGCCACTGGCATGGTGGTGTCCTCTGGGCTGGCTGAAAGCCCTCCTCCATCCTGCATGTC TGCATGCTATTTTTGGGAGCTGGATGGTGCCCCCAGGAACAGGGAGAGGAGTCCAAGCT GCTCTGTCCTCCTGTGAGGCCCCAGCACCATGCCCAGTGGCTCTGAACTGGTGGAAGGG CAGAGTAGGAGAGGAGAGAGAAAGCTAGAAGCTATGCGCCCAGGCTGGCCATGCACGG TTCCTCTGAGAATTTTCTCCCTCCCACTGAGGGACAAGCTTCTCCTGGGGCTCAGGTAGG AGCTCCTCAAGTTCTCAAGTCACTAGTTCTGGCCGCTCCCCTGGGGCTTTGGGTAACAAT TGTCAGCCCCACCCATCCTAGGGTTAGGAAGGCTATTCTCTTTGGCCACTCTCATCCTAA GACCTATTTGGAGAACCTCTGGGGTTTGAGTCTTTTTTTCAGCAGAATGAGGCTTGATCC CGCATTATAGCACCTCGCACATTTGATGTCTCTTCTTCTCACCCACTCACCCCACCCTGGG GGTTGGGGCAAAAAAGTGGCTCAGAGCTGCGGTTCAGAGCTCCTTGTAAACAAGGCTC CTCCCTCACTGTCCTCACCCTGCTCCAGCAGAGGGAGCAGCGGAAGGACCACTCTGC TGCAGCCATGCTTGTTTCTAACCCAGCAGAACTGGACATAATGGGAACAGGGTCTG AAGACAATCAATCCAGGGCTGCAGTGGGTGCTGAGTCTGGGGAAGCCTCCACCTG GAGGGGCAGCTGGGCAGTGGCAGCTCCCTTGGAATGGCTCAGCCTCTGGACATCA CCCCACCCAACCAGAGCCCTGGCTCTTGCTGGATGTCCACAGATGAGTGCCTGGG ATTGGTCTCAGCCACTATGGGGGGGATGTGCAGGGAGAGGTGATGAGGGAGTGAG CAGGACTGTCTATGTGCCTCTGTCCTCATCCTGAGGCTTGGGTCTGAAATTGGTGC TGCAGCACTGGCACGCGTGAGTGGGGCAGGAGGTGGGCAGCAGGGGAGGAGCAT TGGAGGGTGGTCCTGAGCATCATTTGGATTCTTCTCCCGCCTCACTTTAGCCTCAC TCTCACATCACCCTGTCCATTCTCTCCCTTGGAAGCCAACATCAGGAAGGCTGTGG AGCTGGCATATTGCACTATAACAGAGGCTGCTGTTCACGTTTCAAACAACGTGATT GTGTCCCTGGAGCAGTGCACTGTCATGGCCCCGGAGGGTTGGGTCATAGTAGTTAT TAGCTGAATGCCTACACTTCACACTACCTCTGTTCCTCACACCTACCCAAGAAGAG CAGTAATTTCACAGCAAAGAGAGCTGAGATGTGGGGAGGTTAAGTGACCTTGCCA AGGTCGCAGGCAAACGCATGCTGGAGCCAGCATTCAAAACGAGATTAGTCTCACA CTCAAGTGCTTGCTCTCTGCCCCCTCCCTGTGATTACATGCTCATTGTAACACCTGA TGTTGCCTAATGCCTTCCCCTTTTGTTAATGTCAACTGGAGCCTCGTAAAGACCCTG AGTAGCCAGGATGTGTACACTTTATCTCCATGAGACAGAGAAAGGACTGAGGTTTG GAGATGTTAAAAAGACTCGCCTAAACACCCTCGGCAAGCTAATGGAAGAGGATCTG AATCTAGATTATCTGTTCCCAGTGCAGTGAGAAGATGATCTCAGAATGAGTTCCCTG GGGGAACATTTTTTAATCGGAATTTTCATTGACATAATTAAAGACTAGTACACAGTT GTCTTTAATAACGAGCGTTTACACACGGTGCCTAGTTTCTCCCGAGGGCAACATTT GCAAAATTAGTACTATATCAAAACCAAGATTGGACATCAATACAATCAATCTATGATC TTATTCAGATTTGCCCACAATTGCACCATGTGTATGTGAAACTCTAAACAACTTTAA CCTGTGGAGGTTCACGTATCCATCACCACAGTCTACATACTGAAAAGATCCCAAAC CACAAGGATCCCCGCTGTGTGCTTTGATTTATTTATTTAGAGACAAGTTCTCACTCA TTTGTCCAGGCCAGTCTCTTGGGGTGCCCAAAGACCTTTTGTCCAGGTCTCACTTTTGTCCAGCACACATGGAATGGGTGTGGTGGCTCACGCCTGTAATCTCAGCACTTCG AGAGGCTGAGGCAGGAGGATCCCTTGAGCCTGGGAGTTCGAAACAGCCTGGGCA ATGTGGCAAAATCCCAGCTTTACTAAAAATACAAAAATTAGTCGGGCATGGTGGTG CACAACTATAGTCCCAGCAACTCAGGAGGCTGAGGTGGGAGAATCGCTTGAGCCC AGAAGGTTGAGGCTGCAGTGAGGCTTTGTGCAGTGAAGATTTGTACAGTGCTGAA CAGATCCTGTGGAGTGGGGGTGGGCAGAGTGGATGGAAAGGTTGAGAAACTGCA GACATTGTCTTGAGTGTCACAAATGGAAATGCAGCATCCCAGGCAGTATTGCCTGT TCAGGGGCAAGGAACGAAGTGGACATCAGCAGGTGGCCTTGCTGCCATGTGGGTG TCACTATCTCCCCCAGCAGGGGAGAAAACAGGCTTTTTGTTGCAGGTCACACAGCT CATGAGGGGTGGAACTAGATTCACCCTAGGCCTCGCTGATCTCTGTACAACGGGGG CCAGAGCTCTTCTGAGGAAGGCAGGCTTCCCTTTGTACCTGCACTGACTTTTTTCT CCTTTTGGAGGGAGAGCAGAGACCATGTCTGACATAGAAGAGGTGGTGGAAGAGT ACGAGGAGGAGTGAGTATCTGGAGCATCTTGCCTGAGTGGGGTCCTCTCCCGCCG CTGCCCTGACACCTGGTCCAGGAGCCTCCCAGCTGTCCCTCGGATTCTGGGTAGAA GTAGCTGTGTGTGTTTTGGGCACCCCGAGGAGAGATATAGCCATTTCCTTTCCACTCCTG TACCCCTGGGTTGTAGATCCCAGGCAGAGCACAGGCCGAGAGACTTCGGGAAGCACAG AGGTTCAGCTCCAGGGTCATCAGTGTCCTTGGCCAGTCTTGGCTAGGGGCCATGAACAG AGAAGCTGAGATGTTGGGTCTCCTGAAACTGACTGAGGAAGGTTCAGTGACTCTCCAGA GGCACAGAGCTGATTATGCCCCCCAAACCCCAGCTCAGAACCCAGTGTTATCACCCTGTC TTGGATCCCTACTCTCTCATGACAGTGCCCTGCCCACAGTCCTATCCCAGAGGGAAACCA GGGGGTGGGACACAGCCAATCCAATAACTACAGAGGATAGTGGTCACCAGGGAGGAGA CAGAGGTATAGGCACGGCTACTGCCCTCACAGAACTTACTCTTGTTGAGATGTTAAGACA TAAACTGGGAGAAGAAATTAGCAACTGGGGGCCTGGTGTGGAGGGCGGGCTGTTGGTA GTGTAGGAGTTCTGAGGAGAGGGAGATCAGGGGCTGGAATAGTCAGGGGGCTGTGTGG AGAAGGCAGGTCTTACCTGTGTAGAATTTGGAAAGATAAGAGATTTGGGAGAGGGCAGG ACTTTCTAGAAAGGGGTACAGCTTGGGCAAAGATGGAAAGAGTAAGTGAGCAGAACGC TGGTGAGACCAAGCTGGCTGGGGCGGAGGCTTGCGGAGGATGGTAGGAGGCAGCCCTG CATGAGAACGGCAGGCCAGGCTAGTGGGTGTCATTGCAAGGTGGGCAGGGCAGCGTGG ACTCCACTAGGCAACAAGGGAAAAGAAAGGGGGATTATCTTTGGGGAAAGGCCAGTGT GTGCATGTGTGTGCAGGCGTGTGTGTTTGCATGTGCTTGTGTGCGAGCTACTGACAGTGT TTCCTGTTGCTCTCAGGGAGCAGGAAGGTAAGCGTAAACGTGTGTACTCATTTGGAT CAAAGACAGCCTGGTTCGAAACTGACCCACCTCTTCTCTCTTCTCTTGCTGCCTGG A[CTTCT / CTTCTCTTCT]GAGCAGAAGCAGCTGTTGAAGGTATGTGCCTCAGGCAGGT GGAGAGCCAGAAAGGGGCAGGCTCAGCAATGTGGAAATCCTGGGAGCCCTGGGAAAGC AGCTCATCTGTCCCTGCTCCTCACTCTCAGCTGAGCAAGATCCTCCAACAACAGTGCCTT CTAAAATGGGAATCAGTCCTACCTGGGTTGCTCGGGTCTTGAGGCAAACGATTTAAACCT GTCTCTTCTGTCTGCCATTTGCTTCAAACACACTGATCACTTTTCCCAATTTTTTTTGCTTC TTCTCCCTTTTATCATGGGAAATTTTAAATGCACATAAAGGTAGCAATAATAATAATAATGG CCCCCCCTGCCCAAGGTACCCATTACCAGCTTCAACAATCATCATCTTAGGGCCAGTCTT CTTTCCTCTATACTCCACCTTCCACCAGTTATTTTGGAGCAGACATCCAAAATTGTTCCCA GGCACCATGTCAGTTCACGCTTCAAGATTTCGCTACACCGTATCTCTAAGAGATAAGCAC TACTTAAAAACCATAACCACAATGCCATGATATGCCCCAAAAGATTCCATTTAGTAATTTC TCAAATGCGATTGTGAGTATGAAATTGCTCAGTGATCTGAGACGTCTTATCTCGGCATGA AGATTTGTCATGATTCTGAGCTTGCAGAGGCCCCTCTTGCTGCTCCTATGCTGATCGGTTA ACCCATCGCCCTCCTCAGCTGTGGGTCCCACATCTCCCTGAAAGAACGCCCTCACCTTCCCTTCTTCCTGAGAGCTTGGCACATGGAAATGACTCAGAGGCTTCACTCCAGTCCTCCCCC TGCACCCCAGGCAGGTGGACCCTGCCTTTTAGTGGCTCAGGTATCACTTCAGCATGTGGC ATGTAGATATTAGGAGAAGGGTCTCTATGAAGGTCCAAGGGGGGAGCCCAGGTGGCCCC GCCCTTGCTCCCCATGGCTCCTAGGGAAAATGGGGACCCTCCTGCACATTTAGGGCTGTT CCAGGGCTGTCTGGCCAAAGAGGAGACAGAAGAGAAATTCACAGGAGAAACTGAAAG AGGCCACAGGCGTCTTTTTTGGTTTTTAAGGAACACCAGATACTTCAGAAGCTATCAGAA ATATATTTTTGAATAAAAAATCTAACTGGCTCCAAATTCTGCTGTGTTCCCTGAACAGACT CTTGATATGATATCCCTTCCTTTGTTTTGGAGGATTTTCCACTTCCTTATAACAGGTTTAGG GTGGTGGAGTTTGAGACATAGATCTCTGGATTTGGGACCAATCTGCCTGCCACACACAGT AGAATTTGGCTGTGAGTTAGCAGAGTCATTCTCCACTGTGCTGGGGCCTGAGCCGTGTA GCATCTGCCTTGCTTGCCATGAATAGACGAGACTTGCCTCCCACACAGTTGAGGATCTGG CTGCAAATGATCAAGAGAGAGCAACGTCTAGATGGGTGTCCACTAAGCCTGCAGGATAG ATAATGCCAGGTGTCCTTCTTCCAGGAGAGGAATTGTTCCACTTGCTGGAACTGTTTTTT TGGGCAGATATTTGTACGTAGAACAACTGTTGTAGATGCAGCCTCTTTCCTTAGATTTATA ATGCTGGACCCTTCTTAGATGGGGCAGCATCCTCCCAACAGGCCGGGCAAATCTCCCCTG GGGCCAGGCTCTCCAATGAGGCTTTCTAAAGTGTGACTCCTGGAAACTGGACATAGTTT GCAGGATACCCAAGAGCCTCATTCAGAGCAGGAACATGGGCAGGCTCGTTGAGGGCAG GAAATTTGATGGAAGAGAATTGTCCCCCACTCTCTGCTCTTTAGGGGCCTCTGTCTTAGT AAGAAGGAGGCCAAGGCTCTGTCGTCTAGAGGCATTGCAGTTACCACAGATCGCATGCT CCTCTTGTGCCAGGCACCTGCTGGGCGCTCTATGCACGCTAGGTCATTCAGTCCTCACCA CGGCTGAGAAGTGAGTAGAATGATTCCCATTTTACTGATGAGGCACCTGAGGCTCAGGG AAGTAACTGAGAGTTGCCCCTGTTGCTGGAAAGTAGTGGCTTGGGAATTGGAATGCCGG CCTAACTCCAAAGCCCACACTCTTAAAACCACTGCGCTGGGTGGCTGCTCCTGCCGCGG GCTCTCTGCTCCCAGACTAACCTGTCTCGCTTTTCCCCTCCGCTGCGGCCACTCCCTGAA CCTCAGAAGAGGAGGACTGGAGAGAGGACGAAGACGGTAGTACAGCCTTTCCTTCTG TGGTGCTTTCTGCTGCCTGCTGTCCCAAGTGCAGCCTCCTTGTCCAGGGGCCCTGTTCTG GGGGCTGGGGGGTGTGAGTAGGCGGCAGGGACGGAGTGGGTCAGTCGTTTCCTCCCCT GCCTCCCAGGGGCCAGGATCACAGTTCCAAGCTGATCATCATGCCTGAACTTCAGGGCC ACTGGCTCCTCTTCCTGGTTCCTGGATCCTTCCCCTGGGTTGGGTGGTGGGAGGGACCA AGGCTAGCTGGTCTCCCAAGCTGCTCCAAATGTGGCCATAGAGGGGACCTGGCACCTGG AAGACAGCAGCGCCACCCTGCCCCCGCGCACACCCCAGTGTTCCATGCTGGGCCCTCCT TAGGAGCTGGACAGCACCAAGCAGGGTGGCCAGGTGTTGGTTGGGGGGTCTGGGGACA GAGTCCTCTGGAGAGCAGCCAGGGAGACTGGAAATAGCCAGAGCAGGAAGGACATGAC GTCAGCCTTCAGATGCGCCCTGCTGATGGGGAGCACAGGACCAAGGCAAGGGAGTGAG ACCAGGGCTTAATTTTAGAAAGTGCGTTCTGACAGCTATATCTCAAGATGCAGTAGGTGG GTTGGGGAGGCAGTAAGCTCCCCATCCTTGGGATGTTGCATGGCCACTGGGTGGGCTGG GGTATATTATATAGAGAAATGACACGTCAGATGAGGTGGCAGAGGTGTGAACAAGATGG CCTAAAGGTAGACCCCACAATGGCCATGTCCTTCCTCGCTAGCCCTCTGCTTTCACACTT TCTAGAAGGATCATTTTGAGGTAAGACTCCTGCAGGGATCAAATAAAAGGCATCTCCAG GCTTCAGAGAGGACTCAGACCTGGGAAGGGAGGACAGAGGGAATGAAGGCCAACCCTT CTCCCACGATGGTGTCTTCCTCCTGTCTGAGGTTTTTCTTCTCCCTCTGCCAAGAGGCAC CCACTTCCCAGTGCATCAGTGAACAGGCATCACAGAACACTTCCTACAGGCACAGCCCA GGCCAGGCGGCATCCTGTCTCCCTTATCTGTTTGTCTGTCTGTCTACACCAAGTGCCTGA GTGAGATGGGGGCCACATCCTGCCAAACAAATAGATGAAAATAGAAAAATCCTGCCCAC TTTCTGGATAGTACGAAGATCCAGATATTGAGTGTGCCATCCAGCCACACAGTCGTACTCTTTTGGTTTCTTTGAGATGTGGATTGAGGGTGTCTAGAGTCACTAGGTTCAGGACAGGAG CTTCTGGACACCTCGGCTTAAGCAGAGCATGTTCTGTGGTGCCAGACACGGGGTGGAAC TTCTGGGCTGGAAAGTTGGGGAATGAGGTGGAGTCCCATCATCAGGGAGGTTGCCGGGA TGAGTGGGGCTAAGCTGGGTGGCGGGAGGTAGCCGACAGTCAAAACCAAGAATGGCAG CCCCCCAGCACGTTGGTTGCCCCCAAATTCATCTGGACCTAGACGTCTTGACCCCCATCC CCACTCCCCGCTGCCCATTCTCTGCTCTGGGTTCTGCCTGATAGCATGAGTGGGGC CTGCTTCTTCACTCTGTCTGCTTCACTTCTCCAGAGCAGGAGGAGGCAGCGGAAG AGGATGCTGAAGCAGAGGCTGAGACCGAGGAGACCAGGGCAGAAGGTAAGTCTCT GGTTGAGGGTGCAGGGGCCTCAGGGGGTGGCGAGAGCCCCTGGAGCCATTCCTGAGGC AGTAAGAAACCCCCCAGAACAGGCTTTCCCATGTGCACCTGACTCTCCTATTGACTAAG ATTCAACCATTGATTGCCCAATCAACAAACGTTGACCCAGCGCTTCTCTTGTGTCAGGCA GTCTTGGGTCCAGGGGAATGTGTGTGTGAGAGACAGCTCTGTAACCCCAAAAACCAACA GAGTGCATCTTGGCTAGGGCTTATCTGTGGGGAAGGCTCTTGCTGGTGGCGGAGGGGGG GACTTGAGCTCAGAACAAGGACCTTGATTGATTTCTCGTGTTTCATTTAGAAGATGAAG AAGAAGAGGAAGCAAAGGAGGCTGAAGGTAAGGATATCAGGAGAGAGGTACAAGG GAGGCAAAGGAACCCCTCACAGCAGCTTTGGAGGAGAATGCCCACAGTGCTCTTT CCAGGGAAGTGGATGTGCACTGAGAGGGTTTGGGAGAGGAGAGGACAGAACTTG GTGGCACTTCCAAAGTTTTAGATGTTGAGCTTTCAACTTTGCCCCTTCTCCCAGAG AAGTTGCATTGGTGGCACCCAGTGGGAGAGGGGACCATTGTCCAGCCCCCTGGTC TGGACAGAGCAATGGGACCCAGAAGTCATATCCACACATGAAGCTCCTAGAGCTTA GGTGAAATCACTTGTAGGATGTGGTGTCCGTAGGATGAAATCCCAATGCATAGATG GTCATGGTGGGTCATTGCTGCAGTCATCGTCATGCTGCAGCCTTCACCAGGGTTCT AAGTGGTCCCAGTCACTTCTCCAAAGGGGAGGGGACAACCAGCTGGGGCTTGGAA TCATGTCATCAGCTTCTGCCCTGTGGCCACCGGGAAGTCCAAGGGGTAACAAAGA AGGCAAAGACCTGGGAGGGGATTCTAGGGCCAGGACCCTTGGGGACCTTTCCAAA GGCCTTGTCACTGTGAAGCACAAGAGAAGCAAAATTGCTTTGCCCAGAACTGGTGA GTGAGAGAAGGCTTTGTGTGAAGTTCCACACTGCGTGAGTGTGTGCCCTGCAGAACTCT ATGGGAGGGGGAGTGTGGCATCTGCAGCCCCCGCTGCCTTCATTCTCCTGGATGCTTCTG GAATCAGGGCTCCCCAGTGCCGGGAGGGACTCACTGTGCAGATGGGGAAATGGAAATC CACAGGGATCTAGCTCATCGAGGGCGGAGTTGGGACCACTTCTGGGGCCCCCTGA CCCATGGCTGTGTGCTTCTATCTCCTCCAGATGGCCCAATGGAGGAGTCCAAACCA AAGCCCAGGTGAGTGGGGAGGCACCTGGGTAGAGCCGGGAGCAGAGGCTCAGGG AGAGAGGATGATGATGGAACGGGGTGCAGTATGGTGCAGTGGACACGGGCTGGGG GTTTGGTGGGCACAGAGCACCACAGTACAGTTGTACAAGTTGTGTACTGCACAAG CGTCTCTTCCTAAGGGAGTGAGTGAGGGCTGAATTCCACCCAACACTCCATCACCA ATCGTGCACCCTAGCAGGGATGGGGCTATGTTCTTCTAGAGGAAGGGCCCTGAAC GGACACTCTCCTCTCTCACAGTATGGCCAGAACACCCTCCTCCATGGCCGAGGTGG GACCTTGGGATTAAGGGAAGCAAATTGTGGGGAGCCAAACAATGAAACCGTGCCA GCATAGGCATGGCGGCTTCAGAGAGCTGAGAGAGGGCAGGACGGCTTGGTGCCAC ACACGTGAAGCCCACCTGAAGGTGGTGTGTGCTGGGGGGGGCTGGGGGATAGGG AGCATTTCCCTTTTTATGATGCCAACCCTGCCAGCAGAGGTTGACCCATGGCACCT CTGCCCACTCACAGGCTCTTCCCATTTCAGTGGGGCCTTCTGAAACCAGCCCAAGA CATCCCAGATCCATTTATTTGCCCAGTGCCTTCCCCCTTCCTGGCCTCTTGTGCACT GCGGTATCTCATTTCAGCTCCACACACCTGAGGGCAGATGGGGACATGAAGAATCA GAGAACCCCCTGGGTCTCAGTATGTGTTGGGACCAGGACCAAACTGCAAGACTCCCAGGTTCATGGCCAGTTTCTGCGGGGTCCACCAGGGGATTTGAAGTCCAAGAAGC ATGGCATCTTGTCACAGCTTCTTTGATTCCAAGTTGTGTGGCTTTGAGCAAGTCAT TCTACCTCCCTGAGCCTCAGTTCTCCTAAGTGAGATTAACGGTACCCACTTCATGG GCATATCATGAGCATGAAGTGAAATAGCATATGTGAAAGAGCTTTGTGTATTCCCAG GTGCTGAGCAAGGCTGAGGACTGCCATTGTTGACGTCAGTGTTACTATCATTGCTG TGGTTGGTCGGGGCAGTGCTGGAAGATTCTCTAGGAAGGATCAGGGCCCTGCCTG TCCTGGACACCCTCAGTCCCTGGGTCCAGAATGGGGCTGATGCTGACTATTCCTCT CTCCAACAGGTCGTTCATGCCCAACTTGGTGCCTCCCAAGATCCCCGATGGAGAGA GAGTGGACTTTGATGTAAGCGGTGGCTGTGGGTTGAGTAGGCCTGGGCTGGGGATAGT CCCAAGGGCCCTCAGTGACAGAGATTTTGTGACCTTGGGCTGGGGTAGAGCATAGGTGG GATGGGGTGCAGCACCAGCCAGGCTGGGTGCCCATCCATGAGCGTGGTCCTGGGTTCAG GCCACAGTTACCCCTTCCCTGGCCAGGCGCCAGGCCCTGCCAGAGGTCTTTTGCACTGC GTTGGGGGGTGTCTAGCCCACCCATCTCTCCTCTGGACTCTTTGGAGTGGCAGCCTCTGA GCCGCAGCGGTCCACCCACAGGACATCCACCGGAAGCGCATGGAGAAGGACCTGAA TGAGTTGCAGGCGCTGATCGAGGCTCACTTTGAGAACAGGAAGAAAGAGGAGGAG GAGCTCGTTTCTCTCAAAGACAGGATCGTAGGTGTCCAGTCTGCCCACCTAACATTCT CTGATGGCCCAGTCTGTCCTCCATCCCTTTTGTGAGGCCCCACCTCCTCCAGACAGCCTT CCTCGGCGCCTCAAACCCAGCTAGGTGTGAGGTCTCCTTCAGAGGCCCTCCAGCATTCT GCACAGCTCCTGCAGCCCTGACTGGTTTCTCGGGTCCCTGCCTTCCCTCCCCATCCTACT GGATGCTCCTTGACTGCCAGAGCTGAGTCTGATCTGTTTCAGGCCTCCCCCAGGGCCTAA TTCAGGCCCGGGCTGTGGACTGATTGGTCACAGGCCTGCTGCGCCACCCCTGCCCTTGC CTCCTCTTCCCCCGCAGATGAAGGGGAGTGTGACCTGCAGAATCCCACATCCTTCTCCTG CAGAGCGTGGAGGCTGCTTTGATTGCTTTTGTTTCTCCCCATCAAACTTGCCTTCCCACA GACATGCTGCTCTGGCTGGAGCAATGACTTGGCCATCTATCCAAGCATATGGCACAGAGC AGCTGCCAGGGCAGGGTGGGATGGAGGTGGGGGCAGAGCCCAGAAGGAGCCTGCCTG ATGTCTCCCAGTTGTGGAGGGCAGTCTCACCATGAGGAATGTTGGAGGTGGGTCACTGC AGGTACCCTGGTGTGCACGTCCGTGGAGCTGGTTGAAAGTGGAGGCCCTTGGAGGCCG GGCACCATTGCTTCAAGACTCTGGAAGGAACCCTCCCCCAGGAATTCCCTTGCCACCCC ATGCAGGTTTCTGTACCTGCGATGTCACCTTCTCCCTATGCACACCTGGGTGGGCTTTGC CTCTTCTCCTTAGGAGAGACGTCGGGCAGAGCGGGCCGAGCAGCAGCGCATCCGGA ATGAGCGGGAGAAGGAGCGGCAGAACCGCCTGGCTGTGAGTGACCCCTAGCCCA GGCCCATGGAGGCCCCCCATGCTCCCAGCTCCTGCTGGGCCCCCAGGCCCCTCTC TATTCATCAACATGAATGAAATCAGTCAAGAGACAATATTGGCCGGGTGCGGTGGC TCACGCCTGTAATCCCAGCACTTTGGGAGGCCAAGGCGGGTAGATCACTTGAGGT TGGGAGTTCGAGACCATCCTGACCAACGGAGAAACCCTGTCTCTACTAAAAATACA AAATTAGCCAGGCGTGGTGGCACATGCCGGTCATCCTAGCTACTCAGGAGGCTGA GGCAGGAGAATCGCTTGAACCCAGGAGGCAAGGTTGCAGTGAGCCGAGGTCATGC CATTGCACTCTAGCCTGGGCAACAAGAGTGAAACTCCAACTCAAAAAAAAAAAAAA AGGAAAAAAAAGAGACAATATCAATATCAACCTAGGAAGTGCTGGGCATTGTCTTG GCTCTGATAATACATCAGCAAACAAGACAGAAATGCCTGCGTCCTTATGAACGTGT TATTTTATAAGACTCTCCAGCCCTCATCCCTTCCCCAGTGCAGACTTCGTGGGAACC GAAGGCCCCGTAGCTCTCTTCCTGTGCAGCTGCTGAAGGCATTTCCAGAGCAACA GGTCTTCCAGGGGCACTCCCAGCTTAAGGGGATGACGAGACCTCACATTTGGGAA TCAGGACCTCATTCAATCACGCAGTCATTTCTCAGTCAGTGGACTCCTGTGGGCCA GCACTGTGCTGGGAGCTACCCTCTCAGAAAGCTCCTTGCTGAGCGGAGAGAAAGCTGAACTCACCCATAAAGACCACAAGCTTCAGCCCAGAATCAGGGTTTCCAATCCTT TCCCCTAATTTGCTTTCTTCCTCCCTGCTGTAAATCAGGAAGAGAGGGCTCGACGA GAGGAGGAGGAGAACAGGAGGAAGGCTGAGGATGAGGCCCGGAAGAAGAAGGCT TTGTCCAACATGATGCATTTTGGGGGTTACATCCAGAAGGTAGGTAGGGAGCAGCAG GGGTTGCCAGGAGATCCTAGTATAGCCCTGAGGAATGAGGTGTCCACTGCAGCAGGTAG ACTTTAGGTCAGGTCCCAGGAAGAGATTCCCAGCTGCTGTGGGTGGAAGACTGAGGTCT CCTCATGCCCAGCCCTGAGCTGGTTTTCAGCCTGACCGCGGCTTGCACTAGGAGGGGCT TGCTGGGGTAGGGGGCTACAGGCAGAGCCTGGGGCACCAATGGTGCCCCCCATGCCTGG CTCCAGCAGTGCTCAGGAGCCCCCAAGGAGGCTGGGTGCAGCTGGAAGAGCCTTGTGC CTAGAGTCAGAGCTCTAGTTGGAGACCCAGCTCTGCCACATACAGTTGTGTGACCTTTTG CAACCTCTGAGCCTCGGTTTCCTCAACTGGCACTCATATTGCCTGCCTTGATGCTTTGAG AAGCTCAAATGAGCTACCATGGAAAGTGTCTATGTGCCACACAGATGTGAGGTTGTTTGT TCCTGGAAACTCACTCTCGATCCCTAACTTTTGTGCCCCATTCCTCCTAGGTCCTCCATTC TCGAGGGAGGAGGGATATGTGAAATTGATTTTCTCTAGGTAGTGAAGGAATCAAAACATT TTCTTAGATTTCCAGCTTAGTCCCTAGCTCAGGACTAGGCCACCCCCTGCCTCTGCCCCC ATGGTGACCACACCAGCCAAGACTTTGGCATCAAAGTCTTCATGCTACCGAACAGCCCC TGCTGTAACCCTCAGACCCACGTCCTAGTAAACCCGGCTGACTACAGCGAGTATCTGAAT CTTCTCTTCCATGTCTCTCCTTGCCTTTGTCTTCCTGCCTTCTCTCTGTGTCTCTGTTGCCG TCTTGGTCTTTTTCTATGGGCCTCTCTCTCATGGTTTCCCGGTTTAGCAGGCCCAGGTGG GTACCAAAAACCTCAAGCCTTCTTCCTCCCCATAGTTTTGGAGAGCTCCCTGGGAGGGG AAGGCGCCTTGCTGGAGGGGAGGAGTGAAGAGATTGGGCTGGCTGGGGAGGGGGGAA GCTGGTGAGAGGTGAAAGAGATTAGTTAGGCAAAGAAGGAAGAGACTGACGTGATCCT TCTTGCCCCTACCTCAGTTTCCCTAGGGAATACATGTAGTTGGTAGTTGGGGGGTGTGGC AGGAAGAAGAGCATAAGAACCTGGCCCTCCTGCAGGCCTTGCTCCACCCCCAGGGGGT TTGGGGAGGGTTAGGGGACAGGGAGGGGGCAATCTGGCCAGTTTACTCTGCTTCCCACA CCCTCCAGACAGAGCGGAAAAGTGGGAAGAGGCAGACTGAGCGGGAAAAGAAGAA GAAGATTCTGGCTGAGAGGAGGAAGGTGCTGGCCATTGACCACCTGAATGAAGAT CAGCTGAGGTGGGGGCTGGGAGGAATGGTCCTGGGGCACTGCCCGCATCTGCTCTGGG AGGGGTCGGGCCAAGGGCACTTCTTGCTGTGAGCCACCAGAGGGCAGGACTGCCTCAC ATATTTTCCCATGGCAAGTTTGGTGGGACCAGAGCTGGGCCCCAGCAGGTCGGGCCAAC TCCTTTCTGGGCTTTGCTGATGGGGAAAGAGAACTGAGCAGGTGCCCCACCCCTCTGAG CTTCACCTACCCACCTGCACACGGGTGTCAGGACAGGGGATTGAACTAGATGCTCTCCT GCGCCAGAAGGGCTTCTCCTGCTCCCCGCTTGTTCTTGGAACTTTGCCCTGGCAGTCCCT GGTGTCCGTTCCTGAGCACTGGGTGCTGCCGTCTGGTCCCCAGGCTGGGTCAGGCCCAG GGGGTGGGCTGGGGTACCCTTGGCAGGCCTGGAGGTGGGAAGGAGGGCTGGGATGCTG GGGGCTGGGCCGGGACCAGGACGGAGGCCCAGGAGGGCCCTTTCTTACTGGACCTCCC CATTCTCACCCTTGCCCGTGCAGGGAGAAGGCCAAGGAGCTGTGGCAGAGCATCTAT AACTTGGAGGCAGAGAAGTTCGACCTGCAGGAGAAGTTCAAGCAGCAGAAATATG AGGTGGGCCGCCATGCTGTCCCCGCCCAGCATCTCCATCTCACACTCCTCCTGGTT CACTGGGTCCGGTAATACTGCTGCAGGTCCCTGGGTCCCTGTCCCTATTCCCCAAC AGCCCCCTTCAGCTCCTGCATCTGCCCCTGCTGCCTGGCCTTCAGCAGTGTTCCAT GTCCACCGCATTGACGACTGCTTGCTGGAAGTGTCTGAGAGCTCGCTGGGGCTGA GCAGAGACACTTTCCTGGTGTTCCAACCCTGGGGGTCTCCAACACGTTGAGGCAG CAGCTCAGTGATCTGAGCTGGTTACAAGGACCAGGATGCACCAAGCCAGGACCCC CAGTGGAAGGGGAGTGCTGCCAACAGAGAGGTGCTTCTCCCCACATACACCCAAGGTCCTGGTGTGGGCACAATTAGGCTGAGCCTCAAGCTCACAGTCTTTCGGAGTCCT ATGTGCACTAATGAGGGTCTTAGGTGAACAGACACCAGGCAAGGAAATGGCTTAGA GGACACTGATGCTGCATACCGGAGCTTAGACCTGGGCGCCAGTCCTTCCTTACCCA CCACCCCCGAGCCCCGGTCCCAGGGCCTCTTTGCTACCTAAGGGAAGAATTCAGC TTCCCCTGGAAGGTCTCCTTTGCTGCTCCTGCCAAACCACTCCTCCCTGGGCAAGA AGCCCCTCGCTGGGCAGGCTCAGGCTGCAGTGGCTACAGGGTACTCCCACCTCCC AATACGGAGAGAGGCTGTATTGCCTGGTGACAGTGGCATGGACTTTGGAGCCATAA TGCCTGGGTTGAATTTCTACCTGTGCCCCTCACTGGCTGTGTGACATTGGGTGAGT TAGTCCACTGTTCCTTGCCTCCATTTCCACAGGTAACACTACAATATACTTCAGAGGGTG ATTGTGAGGGTTACAGAGATAATACTAATTGTTATTATTGCTATAGTGTTCCAACCACTGTT CCAAGCATGTCCCATGTATTAACTTACTATGCCCTCATAGCAGCCCTATGGGTTCATATCTG GGAAGGTGCTCAGGACAGAGCCTGGCACCCACTAAATGCTCAGCAGGTGTCAGCCATTG TTATGGCCTCTCTAGTCCTGTGCCTTCCACTTTTTTCTCTTTTTTTGGTTCCACACTGAACT CTGCACCGGCCAACAGGACACAGATTTGCCAAACTTTGGGGCAGCACCCTGCAGGGGT GGTGCATGGGGATGCTACTGCTCAAAGGGCACAGCTTCCGGGATGGTGGGCAGCTGGGC AGGGGTGCCCCAGAGGGGTCTGGGGCTGGGCTGCTAGGAGGGCTCCATGACACAGCCT CCAGCTTTGTGCCCAGCTCTCAGAGGCCCTTCTTATGGGACTCTCATATCCTGAACCTATT ATGGCCCTGGGACCCCACAGTGGGAGGCCCATGAGGCATCCTGGAAGCTTCTCCTTGGC TTCTGCCTGTGGTACACGGGCCCCTCCTGACCCTTAACTATCCTAACCCCTCCTACTCTTC CATGCTCCTCCTTCTCCTCCTGCACTGCTGCACTCAGCCCCCTTCTCCCCATCCCCAGGCC ACCTGGGACCTGAGCCAGTCAGCTCCAGCGTTGCTCTTTGTCCTTCCCACTTTTCTTGCA GATCAATGTTCTCCGAAACAGGATCAACGATAACCAGAAAGTGTAAGTGTCTGAGGT CATTCTCGCCTAGCCCTCCCCCTGCCCCCTCCTTCCCTACCTTCCAGCTATCCCATTCCTC CTTGGAGGGCCCCAGGCTCCCAGCCCTTCCTCCTCTGCTTCAGCCCACAGGTTTCCTTCT CTGGCTCCTCCGGGGCCTCGTGCTTCTCTCCCTGTGGCCTCTGCTTGCTGGTGGGGGTGA AATGTGGGGCGGAGAAGCTTCCCTCCAGCCTCAGGCTGCTGTCCTATTCTGCCCCTGGA GGCAGCTCCAGCCTGCTCCTCTCCCCTTTGGCACCCCAGTCCTACCCCAGCCGCATGGTG ACCTACTACCCTGCCTGTGTCTCCATGTCACTGCGTCCTGCTTCCCCTGCAGCTCCAAGA CCCGCGGGAAGGCTAAAGTCACCGGGCGCTGGAAATAGAGCCTGGCCTCCTTCAC CAAAGATCTGCTCCTCGCTCGCACCTGCCTCCGGCCTGCACTCCCCCAGTTCCCG GGCCCTCCTGGGCACCCCAGGCAGCTCCTGTTTGGAAATGGGGAGCTGGCCTAGG TGGGAGCCACCACTCCTGCCTGCCCCCACACCCACTCCACACCAGTAATAAAAAGC CACCACACACTGACTGGCA Wild type MYH7 gene sequence with bold exons SEQ ID 221: MYH7 ENSG00000092054 >chromosome:GRCh38:14:23412740:23435660:-1 – indels are indicated by square brackets ACAGCTCTGTCCTGCTCTGTGTCTTTCCCTGCTGCTCTCAGGTAGGAGCGGGAGCT GGAGGCTTTACTCTGGGATAAGGGGGCTCCAGGCTTAGGAAGGGATTCCTCTTGGAAT AGCAAGCTTCATGCAGGACTTCATGCAGAGTACCAGGTCCAGTCACTGGGCACACATG TGCAGGTCTAAACATGGGCGTATGTGCCACAGGAGTTCCTAGGGGAAGATATCTGCAT CTGAGCATATGGGACCAATATGCATTACAGGGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGGACAAGCCTGCATAGTCTGAATGTGAGAAAGGGTATGTACCTCACTGACTG ATACAGAAATATCAAGTGGGAGATGTGGGTGTATGATCACGTCTGTGAACGAGGAGGT TCCTTTGCAGGGAGGGAGAAAGAATGGTGGAGAAGGATTCTAGGAAGTGTGTATGCCT TCATGTGATTCTGTGAGGCCTGGAATGTGTAGCTACCCCTGTCCATGCATGCAGAGGTT CTTGTCTATGCCTCCACGAGTCATGACTGTTATAGTATCTGTACCCTCTGCAGAAGGAA CTCACAGGCACGGGGGTGTAGGATGTTCCAGGTCTTTGTGTACTGAAGAACACGCAGT GATGTGATCTACTTACGTGGCACACATCCTGAAATGTGCACATTTGCTGTTGAGGCATA TATGGCACCCTGGGGTTGGGGTGCGGATTGAGGTATGGGAAGGAAGCCTGTCTTCATGT GAGAGGCATGCCCAAGCAGCAGGCATGAGTGTTCATCAGAGGGCTTGTGGAGGGTGTT TGGCTATGTGATGGGTGTGTCTGTGAGAGGAAGCCCACAGTGTCTTCTCTCTGAGAATG TCCACAAAGTCTGACAACAAAATGGGGACAGGATGGAAAAGAAGTCTCTTTAGAGTCA TCAGACTTGCATTCAAGTCTTGGCTCTCCCACTTGTTGTCATGGGCTCTTGGCCAAGCAA GCCATTTAACCTCGATTTCCTCATCTGTCAAATGGGGTGAAGACTGAGTAGGAGAAAGT GCATTGAAGTGCTTTTAGACTGGAAAGTGCCGAACAAATGTGACCATTAGTTGTCTCTC AGATGGCTCAAAGAAAGGTACCACTAGATGGGTACAGGCAGGGGTGGGACCATCTCGC AGGAGTGAAGAGGATTAGGGTTCAGGTTGAGTTGACACCAGGAAGCTGTTAGAACTAG TTCTTGAGGAAAAAGGGAATAGGGTTTGAAAAACAAGAAGGATGGGACCAGAGAAGC TGACCACCCTTTCCACCTTGTTTTGGAAGAGGGTGGATGCTGAAGAACAGAGCCAGGG AGAAGCAGGAAGGTGGGACTGGTCAGGTTGGGCACAGCCTGCCCTGACACAGCCTCTT CCCTCTCTCCAGGTCCCCTGCAGGCCTTGGCCCCTTTCCTCATCTGTAGACACACTT GAGTAGCCCAGGTAAGAAAAGCTGAAGCTAGAGTGTTGAAAATCTAGTAAGACTGGG CATTAGAGCCCCAAAGCCAGCCTATGGAACTCTAGTTTCTGCCATGTGCTGGAGAGAAT ATCTTGGCTATCAAGAGCTACTTACCTGTGACCAGGGGGTCCAGGGATAGATGAGGGT CAGAGATGGAAATAGTTAGATTTCTGCTCCTACAGGGAGCTCAGAATGCTCTTCCTTCC CCTAGGAGAGCCCCAGCTTGAACAATAGTTAGTTGTTCCTTTTACCCTGGCAGGTTTGC TAGGCTGCTCCTCTGGTTAAGGGGTAACCTCAAAGAGGAAGGGACTCACTGGTAACTC CTCTTGACTCTTGAGCATGGTGCTAGGTTTTGGGGCTCCCACTGAAGGGGAGAGCCCAG GGAGGGAAGGGAAGAATGGGCAGATGGGAGGGCAGCCAGCTTCTGCTCACTCCAGGC ACAGCCATGGGAGATTCGGAGATGGCAGTCTTTGGGGCTGCCGCCCCCTACCTGC GCAAGTCAGAGAAGGAGCGGCTAGAAGCGCAGACCAGGCCTTTTGACCTCAAGAA GGATGTCTTCGTGCCTGATGACAAACAGGAGTTTGTCAAGGCCAAGATCGTGTCT CGAGAGGGTGGCAAAGTCACTGCCGAGACCGAGTATGGCAAGGTGGGTGTCAGGC TGATGTGAGAGTCCACCCTGGCCACCTGTACACCTGGGTGGACAGAGAGGGGTACACC CATGCCCCATGCCCATGCAGACCTGAGGATGGTCCCATTCTCCTTCCCTTTGGGGAAGA ATCCAGACCCTCCAAAGAGCCCTCTCCTTCCCAATATCCCTTCTGTCTTCCCTGTGAGAT CCTGGTTCCTTCTCTCTTGAGCACTATTGCCCTGTCACTCACCAACTCCTAACCCTCTTG AGGAAGGAGGGAAAGCCCAGGCTGACAGGAGGGCTTGGGTGGGGGCTCTTGCAGACA GTGACCGTGAAGGAGGACCAGGTGATGCAGCAGAACCCACCCAAGTTCGACAAAA TCGAGGACATGGCCATGCTGACCTTCCTGCATGAGCCCGCGGTGCTCTACAACCT CAAGGATCGCTACGGCTCCTGGATGATCTACGTGAGTGCTGCACCTGGCCCTACGTT GGGATCTCTGTTCTTGCTCCATCCATGTCCACCCCAGGAGCCAAGAGTGTCTTCTGTTTG TGTCTAGGCAGGGTTACACTCTAACCTCGTCCCAACATCCTTGGTTCAATTCCAACACTC TGGGGACTGGCATTACTCAGATTGAGTGCATGCAGAGGTTTTCCTTTCTCTTCTTTCTCT CCTGGGATCTTTCTCTAACTCCCAAAATCACCAGCCCTCCCCCTTCGCAACTGGCAAGT CACTGCTCCTTTTCTATCCCCAGACCTACTCGGGCCTCTTCTGTGTCACCGTCAACCC TTACAAGTGGCTGCCGGTGTACACTCCTGAGGTGGTGGCTGCCTACCGGGGCAAG AAGAGGAGCGAGGCCCCGCCCCACATCTTCTCCATCTCCGACAACGCCTATCAGT ACATGCTGACAGGTGAGAGGCCCTGGAAGGGTCTTCCTGAAGGGAACTGGGATAGGC CGGGAGGGAGAGGGAGAAGGAAGGGAGAAGCCCCACGAGAGCATCCTGTGCAGCTCCTGACCTTTCCTCCCCACCCTCTCCCCACAGACAGAGAAAACCAGTCCATCCTGATCAC GTGAGTGTAGCTGCTACTGTATTCCCTTTCAGAATCTCCCTGATCCCAGCCTCCAGCCCC ATCAGGTGTCCCAGGCCCCGAGGCATAGACTCAGCCTCTTTCCCCCCAACCTCGTGCCC ATCTCCTTCTTCTCTGACCATTACCCTGACCCCTCTCTTTGCTCTCCCTCTTCCTTTCTGC TTTCCATCCCCTTCATGTTTTTCCTCTCCCACCTCCTTCACCCACTCTCCAACTCATCACG CGTTTATCCAGAAGCTCATTATCGCCATGTCTTCACCTCTGCATTTTCTTTCCCTTCAGA AACATTTCCCATTCTTCCAGCCCTACCAACTTTTCCCAAAACAGCCCAGGCCCATCTCCC CTGTTTACTTTGCCCAGCCAAGCACAGGCTCAGCCTGCCCAAACCAGTGCCTTCTGGTG GCATCCTTCCTGGTGCCAGCCCTGACCTCTGTGCATCAGAAGACAGTTGTGGATTTTGG GTGTAGATAACCTGGCAGCTTCTTGCCCAGGGCTCCTACTCTCGGGTAGGCCCAGGCAT TCTCTCCTGATTTGAGGCTTGCTGGTCTCCAGTAGTATTGTTCACTGCCCAATAAGCCCC TGTCTTCACAGCGGAGAATCCGGAGCAGGGAAGACAGTCAACACCAAGAGGGTCA TCCAGTACTTTGCTGTTATTGCAGCCATTGGGGACCGCAGCAAGAAGGACCAGAG CCCGGGCAAGGTAGGCCTGCTGCCCTCCAAGGTCCTGTACCGCAGAAAGGGAGGGAG AAGAGCTCTCACCTGCCTCCTTCTTGGCCTCTGCAGGGCACCCTGGAGGACCAGATCA TCCAGGCCAACCCTGCTCTGGAGGCCTTTGGCAATGCCAAGACCGTCCGGAACGA CAACTCCTCCCGCTTCGTGAGTGGTCCCTGACCTTGGCCTTGGGACTTGGACTGGTGG AGGAATGGTCTCAGATATGAGCCTTCCCCCAACTCATCACCACTCTCTTCCATCTCTCCA GGGGAAATTCATTCGAATTCATTTTGGGGCAACAGGAAAGTTGGCATCTGCAGAC ATAGAGACCTGTGAGTGCCATGAATCTGCTAGGCTCAGCCTAAGCTCACCCTTGCTCTA GACCATCTGGTCTTGACCTCTCTCTCTCTCCCCTCCCTCCCTCTGTTTTTCTCCTCTTTAA GTCTCTGTCTGTAGGTGTCTCTGTCTTCAGGTCTACATATCTGTCTCTCTCTGAGACTTCC TCTGCATCTTTCTCCATTTCTGTCTCTGCATGGCTAGGTGTCTTTCTCTGGGATTTCTCTC TGAGACTATTTCTCTCCTTCTGGGTCTCTGTTTCCATCTCTCTGTGTGATCTCTTTGTGTC TGTCCAACTAGTCTCTCTGGCTCTTCCCTTCCCTCTGCCTTTTGCTTGCTACATTTATCAT TAATTTTCCTTGTGCCCAAACCCTAACTTTTCTTTCTCTCCTTCTTCTCCCCACCTGTTCA CAGATCTTCTGGAAAAATCCAGAGTTATTTTCCAGCTGAAAGCAGAGAGAGATTAT CACATTTTCTACCAAATCCTGTCTAACAAAAAGCCTGAGCTGCTGGGTGAGTCAGA GCCACCGACTGAGACCAACTATCTCCATGGCAACCTGGTCCCCTCTGCTTGTGGCTGGA CTCAGCTGGCATGCCCTGGGTTTCATGGCACTGCTGGATTCAATTCAGTGGGATGTGGG GCCAAGCCAAGCCCTGTCCTGTGCTCTTCCTCAGGCCATGTGCTGTGGCGAGCAGCCTC CATGAGAGCCGGGGGCTTGTGTCCCACCCTAACCATGTTTTTTCCCCTAGACATGCTGC TGATCACCAACAACCCCTACGATTATGCATTCATCTCCCAAGGAGAGACCACCGTG GCCTCCATTGATGACGCTGAGGAGCTCATGGCCACTGATGTGAGTGTGTGAGGACC CAGCCAGGGGGTGGGAGGATTGGCAGTGAGGGGCAAACAGGGGTCCAAAAGCAGAGC TAAGACGCTGGCCATTGGTTGTTTAAAGTGGTATGGACCTCCTGAGAGCCAGAATGTGC CCTGTGATCAGGGTGTACTGGGGAATGGATGAGGTGAGACAGGACTGGCCAGTATCTG GTGTTCTATCAAACCAGGAAGAGCTGAAAGCCATGAGACAGAGAGACACACAAGCCA AGAAACAAGCATCACCGAGGGAGCAGGACGAGGAGGGAAGGCCATGCAGACGGCATG GGGCTGGAGGGGCTGTTAGGTGAACAGATGCAGACAGGTATGGAAGGCCAGGCAGGA AGCAAGTGTAGGGCCAGGAAGCATAAGTGGGTAACTCAGAAAAGCTGTGGCCACTGAC ACTGGGGCTGCGGCCAACTGACGTCTGCTCTGCTCTTGCATCTTACATCCTTTGTTTTGG AGTTCCCTGCATCTTACATCCTTTGTTTTGGAGCTCCCCACTGGGTATGGGAGTCTCAGA ACCCACAGGGATTAAGGAGACAAGTTTTCTCTCCAACTTACAAGGGATCTCACTTACCC ATCATACTTCTTTTTCTGGGGTCCGCCAATATGGGGCCTCCCTACAGAACGCTTTTGAT GTGCTGGGCTTCACTTCAGAGGAGAAAAACTCCATGTATAAGCTGACAGGCGCCA TCATGCACTTTGGAAACATGAAGTTCAAGCTGAAGCAGCGGGAGGAGCAGGCGGA GCCAGACGGCACTGAAGGTGGGAGGCAGGGATTCTTGGGGGCAGCTGTCAAGTCATG GAGGGCCATGTCTGCTCAGCAGTCATCTCTCTTCTCTTTTTTCTTTTTTTTTTTTTTTTGAGATGGAGTCTTGCTCTGTTGCCCAGGCTGGAGCGCAGTGGCACGATCTTGGCTCACTGC CACCTCCACATCCTGGGTTCAAGTGATTCTCCTGCCTCAGCCTCCCGAGTAGCTGGAAT TACAGGCATGCACCACCATGCCTGGCTAATTTTTGTATTTTTAGTAGAGATGGGGTTTC ATCATGTTGGCCAGGTTGGTCTCAAACTCCTGACCTCAAGTGATCCGCCTGCCTTGGCC TCCCAAAGTGCTGGGATTACAGGCATGAACCACCACACCTGGCCAGCAGTCATCTCTTT ACCAACTTTGCTACTTGCCTTTTCCTTCCAGAGGCTGACAAGTCTGCCTACCTCATGG GGCTGAACTCAGCCGACCTGCTCAAGGGGCTGTGCCACCCTCGGGTGAAAGTGGG CAATGAGTACGTCACCAAGGGGCAGAATGTCCAGCAGGTGGGTCCATCTTCAGATG ATAATGGGTGGGCAGGGTAGGGAGACTGGCATGGTGGGATGAGAGTTTTCAAGTTCAC TCTTCCCAACAACCCTGCTCAATATGGGTCTCTCCTCCACCTTGCAGGTGATATATGCC ACTGGGGCACTGGCCAAGGCAGTGTATGAGAGGATGTTCAACTGGATGGTGACGC GCATCAATGCCACCCTGGAGACCAAGCAGCCACGCCAGTACTTCATAGGAGTCCT GGACATCGCTGGCTTCGAGATCTTCGATGTGAGTTGGGACCCCTGGGAGTGGGAGA ACAATCACTCACTCGCTCCCACATTCAACAGCTATTTGCTTAGAGCCAGCTGTGGACCA GACATGGGAAGGCAGTGGGGACTGTGTGGTGACAGAGGCAGTCATTTTCTCTGTCTTCA GGGGAAGCCCTCCTTCACTGCCTTGACATGGAGGGGACCAGCCACGCCCTGCTGGGCTC AGGCACAGTGGACGGGCACAGCCCCAATGGCCACTCACACCCACTTTCTGACTGCTCCC ACCCCTCATGCCCCCTGCAGTTCAACAGCTTTGAGCAGCTCTGCATCAACTTCACCA ACGAGAAGCTGCAGCAGTTCTTCAACCACCACATGTTTGTGCTGGAGCAGGAGGA GTACAAGAAGGAGGGCATCGAGTGGACATTCATTGACTTTGGCATGGACCTGCAG GCCTGCATTGACCTCATCGAGAAGGTGCCTCTTTGGCCTTACCACCTGAATTCTCCCT GCACACCCAACAAGAACACCATAGACAAAATAGCAGCCCCTCTCCCTTCTGGGGAATA TAAAAATAGAAGGGGCTGAAGGATCCGAGCCCTTGGTTCCAGAGCCTATGTTGTGCTG AGCACCCAGGACAGGGTGGCACCAGGGACAGCTGATTTCCCAGGGGCTTGAAAGTGGA CACATGGAGGATGGGCACCTGCATGATGACCTCCCACACCTGCATGTTTATTGGGCCTG GTTTATGCATCCAAGGATCAGGAAGTGTGAGGGTGGTGTAGGGGATCATAAGAGTGCA CCTATTTTCAACCCTAGCATCTCAGGCATCTGGGTCGTGGAGTGGTGTGTACAGCATCG ATAGAATCCATATTCCCAGACTTTCACAAAGGTCCTTCTGTCATCAGACAAAATCCTCC ACCTTGAACCGGTTCCAGTCAGTAGATAACTGTACTCAGAGCTGAGCCTACTACCTTAA CACCCAACATGGCACCTCCACGAGCAAGTATATTGACCATAGAGCAGAATCCATGTCC ACCTGTGTGAAGGACACTCAGTGATGCTCTCTCCTGCTTCCTCAGCCCATGGGCATCA TGTCCATCCTGGAAGAGGAGTGCATGTTCCCCAAGGCCACCGACATGACCTTCAA GGCCAAGCTGTTTGACAACCACCTGGGCAAATCCGCCAACTTCCAGAAGCCACGC AATATCAAGGGGAAGCCTGAAGCCCACTTCTCCCTGATCCACTATGCCGGCATCG TGGACTACAACATCATTGGCTGGCTGCAGAAGAACAAGGATCCTCTCAATGAGAC TGTCGTGGGCTTGTATCAGAAGTCTTCCCTCAAGCTGCTCAGCACCCTGTTTGCCA ACTATGCTGGGGCTGATGCGCGTAAGTAGGGACTGAGGCTCCCGGTACAGAGGAGC AGGGATTCTGCCAAGGTTCTGAGCCAGGTCAATTGCTACACCCCACCACTTCAATGTCA GGTTGTACCCCAAGGTTTACAGACTCAGTGGGATGGAACTGGGTGAAGAAACTGAGGC AGCCACATTGAAGGCCCTCACCCAGGGGCCAAATGCCAGCAAGGATGTAAAGAGGGGC TGTGATTCTTTACTCACACCCTACCTCCCCACACTGATGCTTCTTTTGTTGACTCTCCTTC CTGCAGCTATTGAGAAGGGCAAAGGCAAGGCCAAGAAAGGCTCGTCCTTTCAGAC TGTGTCAGCTCTGCACAGGGTGAGTGGGACACAGCCCCAGCCAACTTGGCTCCCCAT CTGCCCAACCCCACCCAGCCCCACCCTTCCCTGCCTTGTTCATCCCCTACTCCTCCCGTT CCCTGTCTCCTTGGTGCATTCGGGACCATTTTCACTCTGTCTTCTCTTCCCGTCATCTCCT GGCCTCTTCACTTATTTACTTCCCATCTCCCTTCTCTCCTCTTTCCCTTCTGTCTCCCACC CTCTCTCCTTATCCCTGTCTGCCCCCAGGGCCCCTTCATCTCTGTGACTTCTCGAATTCTC TCCATCTCTCTTTTCCTTCCTTCTTCTCCTCTCTTCTTCCTGCATCTCTTTCTGGCATTTTC TTTACTTTTCTCTATTGCATTTTTGGCCACAGGAAAATCTGAACAAGCTGATGACCAACTTGCGCTCCACCCATCCCCACTTTGTACGTTGTATCATCCCTAATGAGACAAAGT CTCCAGGTGAGGCCACAAACTCAGGCCAACCCACTGCTGGGCATACACCGCCCTGGGA ACAGGACCTCCTAGGACATCTCCCTACCTACCACCACAGTGGTTTCCAAACCACATTCT TCTCCTCCCATCCCGCATGAGGCCTCCCAGGGTACCTCCCTCTCAGCGCTACCTTGCACT TGCAGTGTGTAATGAGGGTGATGTGTAAGGCACTCTGATGGACATTACCTCATCAGTGG CCCTACAACCCCTTGGATAGCTGGTTGTCCAAGACAGAATCTTAGTTCAACTTTAAATT TACCTGCAGAACCACTTAATCCCCCAATGAGACATATTCAAATTGTTAATCCATTTTTG GCTAATTAGGAGACAGAGGCTCAGGGACCTGGGTTCAAGACCTGGCTCTGTTCTTGATC AGCTTAGTGAACTTGGGCCAGTCCCATAACCCCTCTGGGCTTAATTTACTCATCTAGAA CATTGGATATATCTGTTCTGGCTACCACCTATGATAATAGGTGTGAATCATATAAAAAA GGAATGTGAAAGCCTTTTAGTTAAAAATGTGGCTAGAAGAAAATGAAACAAATGATTA TTACTAACTGACTTGCTAAGATTACAAGCTAATCAGTGACAAAGCCAGGATCAGAACC CAGAACTTCAGTCCAGTGTTCTCACAGACTCCTCCTACTTCCTTCTTGCCACAGGGGTG ATGGACAACCCCCTGGTCATGCACCAGCTGCGCTGCAATGGTGTGCTGGAGGGCA TCCGCATCTGCAGGAAAGGCTTCCCCAACCGCATCCTCTACGGGGACTTCCGGCA GAGGTGGGTATGAGGGTGCACCAGAGCTCATAGAACAGGGGGAGCCAGGCTGCCCTG ATGGGAATGGGATCTGCAGGTGACCCTGGAATTCTATGGGCAGAGCAGATCACTGCAG AGCATGGGTGACTCTGGACACTTCCCTCCTCAGGTATCGCATCCTGAACCCAGCGGC CATCCCTGAGGGACAGTTCATTGATAGCAGGAAGGGGGCAGAGAAGCTGCTCAGC TCCCTGGACATTGATCACAACCAGTACAAGTTTGGCCACACCAAGGTGAGGAAAGG AGACTAATTAATTAAAGGAAGACATCTCTTTTCCATTGACTCCTCTGATGCTTTTCCTGT TGTAATCATCTTAGCAAAATCTCTTACCTGTATGCTACCCCTCCCAGTGGAACATCTAGC ACCACTCCCCTCATCCCAGCTCCAGCTGCCATTGACCTCCTCCCTGCAATCCTTTTCTAG GCTGTTACCCTTCCTAAGGTAATCCCCACCATCTCTTTCCCTCGTACCCCTCCCTAGTCA TGGCCAACACACACCTTGCCTGCAGGTGTTCTTCAAGGCCGGGCTGCTGGGGCTGC TGGAGGAAATGAGGGACGAGAGGCTGAGCCGCATCATCACGCGTATCCAGGCCC AGTCCCGAGGTGTGCTCGCCAGAATGGAGTACAAAAAGCTGCTGGAACGTAGGTG AGAGATCTCAAGAGGAGGTTTCCCGCTTCTCTGAGGCCCAGGCTGGTTCAGGGGCAGT GTCAGGAAAAAAAGCTCAGCAGATCTTCAAACACAGAGACCTGCAGGAGGGGCTCATA TGAACACACTGCAGTCACAGGGTCAGAGGCCTCAGGAAGGGTGGGAGGGATGAAGGA AATGGGATATTCCCAAGGTTTCAGGACCTCAGGTAGGAAGGAGGCAGGAGGCTCAGCA CTCCTTTCAATGGGCCCCTGCAGAGACTCCCTGCTGGTAATCCAGTGGAACATTCGG GCCTTCATGGGGGTCAAGAATTGGCCCTGGATGAAGCTCTACTTCAAGATCAAGC CGCTGCTGAAGAGTGCAGAAAGAGAGAAGGAGATGGCCTCCATGAAGGAGGAGT TCACACGCCTCAAAGAGGCGCTAGAGAAGTCCGAGGCTCGCCGCAAGGAGCTGG AGGAGAAGATGGTGTCCCTGCTGCAGGAGAAGAATGACCTGCAGCTCCAAGTGCA GGCGGTGAGGCTCCTGGGCTACTGTTGGCTCTTCCACCCTGCTCTGCCTTCACTTCCCA CAACCCTGCACCTCCCTGCACAGGGGCTCACTGTCTTGTTCCTTCAGTCAGAGGACTCT GGGATCAACACTTCTAAAGACCTCCAAAGAGGTCCCTCAGGAGGAGGAAAGGGCAGA GGGAAAAGAGCTGACTTTTAAAAGCACAGGCTTTCTACTTGGGTTCAACCTAGATACAC AGCTTAATTATTTTGTGACCTTGGGCAGACCATGCATCATCTTTGAGCCTCAATTTCCTC TAGACAGGGAATAAAGCTGTCCATTTCATTAAGCCATCATACATAAGTGCTTGGCATCA GCAAATGCAATTCCCACCCTTCGATGGAGGAGACCAGAGGAAGAGGTCCAGATGAAGA TTTCTGGTTCCTTTCCTCTCTGCTCCCCTCCCCAGTGTTCCCAAGTTATACTATCCTGAAA CTCTTCCCCTCCCCATACTTCTGAAGCTCTTGCCAAGTGGGGATATCCTAGAGTTACCCT CCTATTTGAGTGATGTGCCTCTCCTTCCCTCTACCTGCAAGAATGAGGACCTTACCCCCT GAACAGCCTCCCCTCTGTTCCTCACCTTCAGGAACAAGACAACCTGGCAGATGCTGA GGAGCGCTGTGATCAGCTGATCAAAAACAAGATTCAGCTGGAGGCCAAGGTGAAG GAGATGAACGAGAGGCTGGAGGATGAGGAGGAGATGAATGCTGAGCTCACTGCCAAGAAGCGCAAGCTGGAAGATGAGTGCTCAGAGCTCAAAAGGGACATCGATGATC TGGAGCTGACACTGGCCAAAGTGGAGAAGGAGAAACACGCAACAGAGAACAAGGT AAGGGCAGCTCCCTTTGGCTCCAGCCCGGGTCTCATCAGGACTCTCAGACCATACTGAC CTTGACCCAGGCTAGCCACTCGGCATCCAGAGAGCAGCATGGACCTTGACATGGAGCT CCCCACAGATGGCACCAAGCTGGTGACCTTTGACCCTAAAGGAGATGGGATTCTTGGTC GGCAGGTGAAAAACCTGACAGAGGAGATGGCTGGGCTGGATGAGATCATTGCCAA GCTGACCAAGGAGAAGAAAGCTCTGCAAGAGGCCCACCAACAGGCTCTGGATGAC CTTCAGGCCGAGGAGGACAAGGTCAACACCCTGACTAAGGCCAAAGTCAAGCTGG AGCAGCAAGTGGATGATGTGAGTAGATTGAGAGTTGTGGGGCCTAGATATGCCATGT CTATCTGTGCCCAGAGCTCTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGT GTGTGTGTGTTTGTGTTTATGTTTAGAGGGGGGTAGGGTTAGAGGATGTTAACTTTTCTG ATATCTTTCTAGAAAGGAGCTGGGGAATGAAAAGTCAGAGGTGGTGATCAATTCTGAA TTATGTATCTAATCTCACTGGTATGATCAATAATAGAGATGAGTTTTCTTATTTAATTTA AGTGCAATCCCATATATCTCTGCCTTTGACACAAGATTTAGAGGGTCATCTCATAAGAA CATGATAAATTTCTTTGGAAATTTTGTGAGGCTCCCCAAGTAGAATTTTTGAAATTCCA ATATTTAAGGCAGAATCTGATCATTGTTCCCTCTGATGCACATATGTCTGTTGACTCACT TATCACTCTGTATTTTAATTATTTAAGCTTCTATCTCAGCCATAAGACTACAAGCTTCCT TTAGATAGGAACTTAGATCTATTCAATTTGGGGACTGCAGTGCTTAGCACAGAGCCTGA TGCCCAGTAGGTGTTTATAGGATGCTGAAAGAATGAATACGGCGGCCAGCCGCGGTGG CTCACACCTGTAATCCCAGCACTTTGGGAGGCCAAGGCGGGTGGATCACGAGGTCAAG AGATCGAGACCATCCTGGCCAATATGGTGAAACCCCATCTCTACTAAAAATACAAAAA TTAGCTGGATGTGGTGGCGTGCGCCTGTAGTCCCAGCTACTCAGGAGGCTGAGGCAGA ATAATCCCTTGAACCAGGGAGGCGGAGGTTGCAGTGAGCCGAGATCGCGCCACTGCAC TCCAGCCTGGCGACAGAGTGAGACTCCATAAAAAAAAAAAAAAAAAAAAGGAGGAAG GAAGGAAGGAAGGAGAGAAAGAAAGGAGAGAAAGAAAGAAAGAAAGAAAGGGAAA GAAAGAAAGAATACGGCTTTGGGGTTTCATCCCACAGTCATTTATTCCTTTATCTATAA AATATTGATTCAGCATCCCCTATATCTAGGCAATCTCACAGTCCCCTAATAACATTTGG CCCACTCTGGGGAAGGTTTCCCAAGTCCTGAACACAAAGATTTACCAAGTCCTGAGGTA ACTGAACAACAAAATCACCAAGTCAGGATGCTTTGCCTCACTGGGCCTGGCTCCTTCTC TCTACCAGCTGGAAGGATCCCTGGAGCAAGAGAAGAAGGTGCGCATGGACCTGGA GCGAGCGAAGCGGAAGCTGGAGGGCGACCTGAAGCTGACCCAGGAGAGCATCAT GGACCTGGAGAATGACAAGCAGCAGCTGGATGAGCGGCTGAAAAAGTACTGTGTC CCCTCCCTGCTGCCCTTCCTCCTCCCCAGCCCATAACAGTACAAGCAGACCCAAGACAG CCATCCTCTGAGGCAACAACCTCACGCAGCCTCCACAAGTGGAGGCGCAAACCATGGG AAAAGGCCTCCCAGGCAAGGGCTTTCCTCCCCTAGTCCCAGATATCAGGGTTAGTTAGA TGGTCCTCCCTGGCAAAAAGTCCATTGTATCCCATGAGTCCTTTCTTCTTGTTAGGCACT CAGTGGGAATGAAAATAACAGCCTTCCCCCTTCATACACATCTCTCTTTGGAGACCAGT TCCTTGACATGGAGGAAGGCTCCACATGAGAGCCTCTTAGGAAAGTCCCTTTGTGGCTT TTCACAGAGAGCCCACAGGGCACCAAGAAATGCCCTCACATTAAAGAAGTCAGGGGAT CAGGGCCATTTCTTTGTTTTGCTTAACTTGTGGTTCCAATTCTTAGTGTAAGTTCTTCCA AAATGTATAGACTTTCCAGACTTGGTAAACAGTGAGCTTGCAGAACATATTTTGATAAT TGAAGGTCTTATGGTGTTTCAGGCTCAGGACTTTGAAAGGCTTTGAGAATCATCATAAT CTACATCCTCATCAAGTTTGTGTTTGCACAGGAATCTTCATACCTTCCCAGGCACAGAA TGTGTTGAATAAATGAATAAATACACGATCCTATTTGATTCTCAAAACACCTTTTAAGG ATGGTATCTTTAATTTCACTTTACCAAAGAGGAAAATAAGTTTCAGAAAGGTTAGCTGA GTTACCCAAAGCAATAGTGACAAAATTAAAGACCATCAGAGCTAAAAAGGACCTTTGA AACAATCTAGTCCGACCCCCTCATTTTACATTATGAGAAAACAAGGACCAAAGATGGG AATGTCCTGTAGAGCCCAGGCCCCATCACCCTGTCTACAGCACTCCTCCCATTACATTCT TGTTGGATTTCAATCTCACCGCAGGTGTTACACTTCCAGGGATCCTACATCACCCCCAGAAGCTGTTTTTCTAATGAAATCCTACTCTTTACCTGTATCATTACCATTTTCAACCACTG GTGGACCCTCCTGAGGCCCCACGAGTCTCCCTTACCTCACCATCCCTCCTTCCCCACAG AAAAGACTTTGAGCTGAATGCTCTCAACGCAAGGATTGAGGATGAACAGGCCCTC GGCAGCCAGCTGCAGAAGAAGCTCAAGGAGCTTCAGGTGAGGTCTGGACACCCACG CGGGATGCTGAGTCCCTGGGCTGGTTCTGTTTCCCCTGCCCCCTCCTCTAGGGTGTCCTG TTCACAGCATTACAGGAACTGATTATGTGTCCATGGAGGCAGGCACGCAGCTATCACA GTGTGCCCATGGATACATCCAAAAGGGGTCAGGTGATCCACAGAGACCCTGAAAAGGT CACAGGCCTGGAGGGCAGAATTCCTGGCTTCCTGTCTCAGCTCTGCCTTAGTTGGGTGA CCTCTGGGAGCCCACTCCAGATCTCTGGGTCTCCATGCCCTCCCCTGTTACATGGAGGTT TTTAAGAGTCCTTCGACTCTGATGGCATGACTTTCCTCCTTACTCTAGAGCAGAAGCTGT GTTAGGTCTTTAGTTGTCTGGTCAGACAAATACGGCTCTCTCATTTTCAACTTTTGTTAT TTTTTTATGACTAAACTGCTTAAAATATCCCCAGAACACAGGTCTCTTTGTCCATCTTTA AGAGCAAGTTTCAACCACTTTCCTTCCAATAAAGAGCATGTAGACTGTCAGGCCCTCGC CTAAAGGAAGGCTCTTCAAACTATCTTCCCCAAATCCTTTAATTCCCACCTCTCCACTGG AAGAGCTAAACTGACTTGCTGTTCCAGAGAAGCCGAGAGCCTTTTAGAGCCGGGGGGA TTCCAGTGGAGGGGTCCAGGCGGTGGGTCTGAGCCCTTTGTGTCTGA[CC / C]AGGCACG CATCGAGGAGCTGGAGGAGGAGCTGGAGGCCGAGCGCACCGCCAGGGCTAAGGT GGAGAAGCTGCGCTCAGACCTGTCTCGGGAGCTGGAGGAGATCAGCGAGCGGCT GGAAGAGGCCGGCGGGGCCACGTCCGTGCAGATCGAGATGAACAAGAAGCGCGA GGCCGAGTTCCAGAAGATGCGGCGGGACCTGGAGGAGGCCACGCTGCAGCACGA GGCCACTGCCGCGGCCCTGCGCAAGAAGCACGCCGACAGCGTGGCCGAGCTGGG CGAGCAGATCGACAACCTGCAGCGGGTGAAGCAGAAGCTGGAGAAGGAGAAGAG CGAGTTCAAGCTGGAGCTGGATGACGTCACCTCCAACATGGAGCAGATCATCAAG GCCAAGGCAGGCTCTGCTCGGCCTCCCCTCCTCCAACTTCCTCCTCCCACCTCCCCTTTC TGTCCATGTAGTGTCTCTTCCTTCATGGTTCATTTTCCCACTTCCCTTCCTCTGCCTGGTT CTCAGATCCCCTCCTTCTTTTCACACCTTCCCTCTTCTTTTCTCCTTCAGTTGCACCTCTT ACACCCCTTCATTCCCCCCGCCCCCAACATCCATCATATAACTCTCCTTCCCTTCTCAGG CTAACCTGGAGAAGATGTGCCGGACCTTGGAAGACCAGATGAATGAGCACCGGAG CAAGGCGGAGGAGACCCAGCGTTCTGTCAACGACCTCACCAGCCAGCGGGCCAA GTTGCAAACCGAGAATGGTGAGCCTAGAGCAGGGGCTCCATGGTTCCCACCACAGTC TCCCCAACCTCCTGAGCCACTCCAGCCTCGGGCACAAGCCAATACACGCACACACAGA CACAGAGTCTCTCCAGAAAATGGGGCCTGAGGGCTAGAGGAGGAGGTGGGGATAGAG AGGAGTGCTGATCTAGAATGGTGCTTCCCCAGGTGAGCTGTCCCGGCAGCTGGATGA GAAGGAGGCACTGATCTCCCAGCTGACCCGAGGCAAGCTCACCTACACCCAGCAG CTGGAGGACCTCAAGAGGCAGCTGGAGGAGGAGGTTAAGGTAAGGGCTAGACTCG GGCCACCTGGCCCAAGCAAGGAGCACACTGACTAGCCTTGCATCAA[C / _]ATCACTTCC CTTCCCAGCAATAAGGCTGGCTGTGGACCAACAGTTCTCCAAGAATTCTAAAAAGAAA TCACAGCCCAAACCCTGCCACATGCCATCTACTCCCCAGGCTGAGGCCCTCTGCTTCTG CTGCATCCCTGGACGGGAGGCAGGGTGTGAGAAGGAGGAGAGAAAATGAACTAGGGA GGCACGAGATGGCCTTGCCCGGGCACGGCTAGGGGATGTAGGGACTGCACAAATGGGT CCTGGGATTTGCAGGGCCACCTGAAAACTGGAGAAAGAAATTCAACTGGAGCCTATTC CTAGCACTTTGCTGGACTCATGAAAAAGGAGAAAGAACATGATTTTTAAAAAACTCCA GGGTTCATACTAACCTGCCTCTTATAGCTCAGTGATCTTAGGTTAGGGAAGTCCCCTAA CCTTTCTGAGCCTGTTTCCCTCTCCCCAGCCAGGTAAAATGGCGGTGATGATGACCATG TCTGGTGTGGGGAGGAGCTCAGGGCCAATGGCCATGAAGCTTTTCTTAAATAGTTCACT GCTGTATTGGCATTCTTTGTTATCAGCAAACAGTAAACCATGACAGGTCCACTGGGGGT TTGGACATAGATGAGGCCAAGGGATGATGTTGAGATTGGGGTGAGGAGAAGGGCAAG GGTGGGGTTGCTTTATGGAGAAAGCTGAACCCACCTCCTGGTGCCCACCCCTCCCCAGG CGAAGAACGCCCTGGCCCACGCACTGCAGTCGGCCCGGCATGACTGCGACCTGCTGCGGGAGCAGTACGAGGAGGAGACGGAGGCCAAGGCCGAGCTGCAGCGCGTCCT TTCCAAGGCCAACTCGGAGGTGGCCCAGTGGAGGACCAAGTATGAGACGGACGC CATTCAGCGGACTGAGGAGCTCGAGGAGGCCAAGTGAGTTCTGAGCAGCCTGACTT CTGGCTGAGGCCCCTTTGCAGGCAGGACTCAGCCCAGCCCCAGCCTCAGCAGATCCCAC ACAGGCGATGCTTAGCTAGTGTTTGACAACACAGGAGGACTCTGCCCCGGCCCCACCTC CTTCTCCTCTCAGGGAAGCTTTTTGCTCGAATTATGTTTCTGATCCGAATATAAGACGAA CAAAAGGTTTGTCTGAGGGCAGAGTGCTTCCGTCTGGGGCAGGGCACTGTGGCAGGGA AAGGCAGTGGGGAGGGCTGCAGAAGCCCATACCTCCTCAATGTCCATAGCGCAGAGGC TGGGCCAGGGTCAGAGGGTGCCTGGGTCTCCACGCCCTGCTGGGATCCTCTGCCTGATG TTCTCGGCCCCTGGGACCTGTCCTCAGGCTTCTCCAGCTACACTTCTGAGGTTTCAAGGA TTGTCTTTGAGAAGGTTCATGTTGTTTCCTCTTGTCCCCATCCACACCCTCCATCCTCCCC ACCCTCTGCCACCCTCCCCTGGGCAGGAAGAAGCTGGCCCAGCGGCTGCAGGAAGC TGAGGAGGCCGTGGAGGCTGTTAATGCCAAGTGCTCCTCGCTGGAGAAGACCAAG CACCGGCTACAGAATGAGATCGAGGACTTGATGGTGGACGTAGAGCGCTCCAATG CTGCTGCTGCAGCCCTGGACAAGAAGCAGAGGAACTTCGACAAGGTGGGCCCTGG GTGGGGGCCGCAGCCAGCATGCAGGGCAAGGGGGCATGAGGGGTTCAGTGAGAGGCC AGAGCCATCCTCCTTGGAGGTGGGGGAGGAGGGCTGAGCCCAGGCAGGTCCTGAGACA GACCCTGGACATGGGGCTGAGGCTGGGGGGCTGAAGAGTGAGCCTTGTCCCCGGGCAG ATCCTGGCCGAGTGGAAGCAGAAGTATGAGGAGTCGCAGTCGGAGCTGGAGTCCT CGCAGAAGGAGGCTCGCTCCCTCAGCACAGAGCTCTTCAAACTCAAGAACGCCTA TGAGGAGTCCCTGGAACATCTGGAGACCTTCAAGCGGGAGAACAAAAACCTGCAG GGTGTGCTGGGGGCCCAAGAGGCTGGGGAGGGGCTGCACTGGCAGTGTTCCCATATGG TGCCACCCAAGGGCTCCAAGAGGCGTCTGGGCAAAGAGGCGTGTCCCTCCAACTCCAC TGGACCTCAGCAGCCCTCAACCGAGTTACCGTGTTCCCCACACAGAGGAGATCTCCGA CTTGACTGAGCAGTTGGGTTCCAGCGGAAAGACTATCCATGAGCTGGAGAAGGTC CGAAAGCAGCTGGAGGCCGAGAAGATGGAGCTGCAGTCAGCCCTGGAGGAGGCC GAGGTGTGTGTGTGTGCAGGGCACGGGGTGCAGGGAGCTGAGCTCCAGGCTTTTGGTC CCTGTTCTCATCCCCCGCTTTGTCCGTTTTGTTCACTGTCCCATCCCCAGAGCCTAGGAC AGAGCCTGGCACATAGTAGGCATTCAGTGAAGATTTGTGGAGTGAATGAATGACCAGT CACTGAACTCTCTTCACATAAACATTTTACCCATTGCATCTTAGCTGAGCTTCCCATTTC CACATACCTCCCGGTTCCCACCTCTGCCTCCCTCTGTTTTCTTTTATACTCTATACCTGAT TGCCTCTGTTTCCTTGGCAACACTTTTCTCTTCATTTTCCTCCTTGTCTTCATAGCTGGCT CTCCCCTGTGAGGGCCTGAATCACTTTCTCTTAGGCTTATTTTTATTTTCTCTTAGATTCT CTTTTCCTCCACCTTCTGCTTCTTTGAACCACTTACACCACTCTTGAAGTCACTTCGTATC CATGATTAGTGAGCAGGCCCCCACCCTGCCCTGTGCCCTGACTGTCTGCCTGCATCCCC TCCCCCAACCCCTTCCCAGGCCTCCCTGGAGCACGAGGAGGGCAAGATCCTCCGGG CCCAGCTGGAGTTCAACCAGATCAAGGCAGAGATCGAGCGGAAGCTGGCAGAGA AGGACGAGGAGATGGAACAGGCCAAGCGCAACCACCTGCGGGTGGTGGACTCGC TGCAGACCTCCCTGGACGCAGAGACACGCAGCCGCAACGAGGCCCTGAGGGTGA AGAAGAAGATGGAAGGAGACCTCAATGAGATGGAGATCCAGCTCAGCCACGCCAA CCGCATGGCCGCCGAGGCCCAGAAGCAAGTCAAGAGCCTCCAGAGCTTGTTGAAG GTACTCACCCAGAGGGGACTGGCCTCCACGTGGCCTGGCGAAGCAGTAGTGTCTTGAT ACAGGCACCAGATTCCTCCTGCCCCTAGGTTACTGCAGGGACCTCTGACAGGTGCCTTT AGTGAAGGGAACCGAGGCTGGCTCCCTGCTCATGCCCACTCTCCTGATCCTCAGGACA CCCAGATTCAGCTGGACGATGCAGTCCGTGCCAACGACGACCTGAAGGAGAACAT CGCCATCGTGGAGCGGCGCAACAACCTGCTGCAGGCTGAGCTGGAGGAGTTGCG TGCCGTGGTGGAGCAGACAGAGCGGTCCCGGAAGCTGGCGGAGCAGGAGCTGAT TGAGACTAGTGAGCGGGTGCAGCTGCTGCATTCCCAGGTGAGCAGCTCCCCTGCTC ATTCCTGAAGGGAGCACAGGCTGGGGCTCAGCAAGCAAGGCTTGAGAGCTATGCATAGATGCTCAATGCTTTTCCTGCTCTGCCCAACCCTCCCCCAACCCAGAACACCAGCCTCAT CAACCAGAAGAAGAAGATGGATGCTGACCTGTCCCAGCTCCAGACTGAAGTGGAG GAGGCAGTGCAGGAGTGCAGGAATGCTGAGGAGAAGGCCAAGAAGGCCATCACG GATGTAAGTCCCCCACTCCACCGACCCGATCCAGACCAGTGTCTCTCCGTGGGCTGGGC AGCAAGTGTGTGAGGACTTGACCAGACCATGTGCCACCTCTCTCCTGCACACAGGCCG CCATGATGGCAGAGGAGCTGAAGAAGGAGCAGGACACCAGCGCCCACCTGGAGC GCATGAAGAAGAACATGGAACAGACCATTAAGGACCTGCAGCACCGGCTGGACGA AGCCGAGCAGATCGCCCTCAAGGGCGGCAAGAAGCAGCTGCAGAAGCTGGAAGC GCGGGTGCGGGAGCTGGAGAATGAGCTGGAGGCCGAGCAGAAGCGCAACGCAGA GTCGGTGAAGGGCATGAGGAAGAGCGAGCGGCGCATCAAGGAGCTCACCTACCA GGTGCGACGGGCGGTGACTCCAGGCAGAGCCCTGGCACCATAGCCACAGTGACAACCA GCTGAGGAGAATGAAGAGTTTGCTCTTAGCCTCTTCCAGGGCGAGGATGGGAATGCAG CCCCCGTTTCACTTTGCCTAGCCCTGCCCCACTCTGAATGTCCCCTAGCTCAGAGGTCAG TCTCTGAGCCTCCTGGCCTGGGAGCTCCATCTCAACACCCACTCTTAGCCCATCGCCAG GGGACACACACAGAATTCCAGACAAAGCTCACCCAGTACAGCTCACCAGAAAGCAAAT ATGTAGCCAGGGTCACCCCCAAAAGACACCAAAAACACACCCATCCCATTCAAACAAG CATAAAAACTTTTTCATTGCCCAGGGCAGTGTGGACTCTAGCTTTCTGAGGTGTTTTCTA GAATCAGACTCTGAATTAGAATTTGTTTCTTTTCCAATCCAGGATGTCACTATCTCTCTA AGCACACTTTGTCCTTAACTTAGATTAAGTCAGTGTTTTCCTGGCCTGCATTGCAGCCCA GGAAATTCCATCACACTCATAGCCCCAAAGTCACTCCCAGCATGTGCCGAGGTCCAGA GCAGGGGACTTCATCCCGCATGGGGTCCCCGGACCCTGAAAAACCTGCTCCCTGGAGC ACTGCACACACACAATTTTGTGCATAATTTCAGGAGTCCCACTAGTTCTTTGGACTCTTT CCTGGTACAGATCATTTAAAATATTTACACATATTTGGTAAGAAATTATAAGGAGAATT CAAGTGTTTAGTGAGGATCAGAAAGTAGAATTGGGTCAGGATATCAGATGAAGCAGGG CAGGGCGGAGGGGATGCTACCTTCTATGACTGTGCCATCTTCACCCCCTGCCTACCCTC TGGCCCCCAGACGGAGGAGGACAGGAAAAACCTGCTGCGGCTGCAGGACCTGGTA GACAAGCTGCAGCTAAAGGTCAAGGCCTACAAGCGCCAGGCCGAGGAGGCGGTG AGTGACCCTGCTGGGGACTAGGCCCAGGGGAGGCATAGGAGAGCTCGTCCCCCAAGCC AGGAGTCTGAGAACCCAGGCCCCCTCTCACCTCATGCTCCCACCTCCCGCAGGAGGAG CAAGCCAACACCAACCTGTCCAAGTTCCGCAAGGTGCAGCACGAGCTGGATGAGG CAGAGGAGCGGGCGGACATCGCCGAGTCCCAGGTCAACAAGCTGCGGGCCAAGA GCCGTGACATTGGCACGAAGGTGGGTCCCTCTTTTGGGCTTTGCTAGTCACCCCCACA GCAGGCATACCCAGACAGAGCACCCTCAAACCCGGGATGCTTCCTTTTCATTTATTCCA CACACTTGAGCCACATGGCCCCAGAGGCCAAGGTAATGCAGTTCTCTGTGATTTCAAGG ATTCTTCCAGCTCTAACTTTTTTTTTTTTTTTTTTTGAGACGGTGTCTCACTCTGCCGCCC AGGCTGGAGTGCAGTGGCACAATCTCAGCTCACTGCAACCTCTGCCTCCTGGGTTCTCC TGCCTCAGCCTCCCAAGTAGCTGGGATTATAGGCATGTGCTACCATGTCTAGCTAATTT TTATATTTTAGTAGAGATGCGGTTTCACCATGTTGGCCAGGCTGGTCTTGAACTCCT GACCTTGTGATCCACTCGCCTCAGCCTCCCAAAGTACGGGATTACAGGCGTGAGCCA CCACTCCCAGCTCAGCTCTAAGTTTCAGTGGTTCTGGGAATTTCAGCCTTAGGGAGGGG TGCCACCCTAGAAGACCCTAATTTATATACAACCAGTATATCCCAGTTTCCCCCCAACT GCTAGTTTCCAGTTCCCTTCCACCTAATTTTGTCCTGCAATCTTGTAACTCAAACCTAAA ATGGACATCCCAGAGAACCCGGGCCTGGCTTTGTTCTCGCCCAGGCAACTTCCCTCTGA CCAACCCCAGAACCCTACCCTCCAGAACTTGCCCCCCGAACATCGATCCCTGGAAACAC ATGTGCTGCTGCCACAGGGGCTTCCACCTGCCCACAAAGCAGGCCCCCCACCATCAGAC CCCTCTCACCTTTGTTCCCATGCCCTGTCCCTGCCCAATACCATCTCTCCAAGGACTGAT TGGACTTTGTTTCCTTTCAAAAGGGCTTGAATGAGGAGTAGCTTTGCCACATCTTGA TCTGCTCAGCCCTGGAGGTGCCAGCAAAGCCCCATGCTGGAGCCTGTGTAACAGC TCCTTGGGAGGAAGCAGAATAAAGCAATTTTCCTTGAAGCCGAWild type TNNI3 gene sequence with bold exons SEQ ID 222: >chromosome:GRCh38:19:55151767:55157773:-1 – indels are indicated by square brackets AGGTGACAGTATATTTAGTCTGTGTCCTCGCCCTTTATCTCAGTGTCCTCGGGG AGTCTCAAGCAGCCCGGAGGAGACTGACGGTCCCTGGGACCCTGAAGGTCACC CGGGCGGCCCCCTCACTGACCCTCCAAACGCCCCTGTCCTCGCCCTGCCTCCTG CCATTCCCGGCCTGAGTCTCAGCATGGCGGATGGGTGAGTGATGCCCCAAGGC AGTGGGAGTTGGGGGCGACCTCCCGGGTTCCCAAGAGGGGTCGCAGCTGAGAGGCT GGACCCTTGGCCTGCGAGGTAGGCGTAGGGACTCTTGGGTGAAGAGAGGAAGTGGG TTTGCGAGTCAGACTCCTGGAACCCAAGGAAGGGGAAGCGCGGTCCCCCGACCTCTT GTTCAGAGGGGACTCCAGGGGTCCCTTAGG[AGACAG / AGACAGACAG]GACACAGC CCACCACTAACCCCCCTCCTTGGTTTCTCTCCTTCCAGGAGCAGCGATGCGGTGAG AGCAGCGGGCTAAGGCGTGGCTGGGACCCCCAGGGCCAGGGTGGGCGCTGCAG TGAGGGGTCTGGGGCGGGAGGCTGCAGCCCTAGCAGAGGGTGCGGTACGGTAAGGG CTGGGTGGGGTCTTGGTGGTGATGGGGTCCCCACTCCTCCTAACCCAGGCTAGGGA ACCTCGCCCTGCACCAGCCCCAATCAGACGCCGCTCCTCCAACTACCGCGCTTA TGCCACGGAGCCGCACGCCAAGGTGGGACGGGGCTTCCTGGGGGCAGAGTACA GGCGCCGGAGGGATCCAAGACCCTGGGAGTGGGGGGAGGAGCCAGGGCTGCG AAGGGGGCGGGGACTACGCGGAGGGGCTTCAGGGGCGGAGTTTTGCAGAGGG TCATGCTCGGATTGGTGACAGCAGCCTGCGGGCGGAACTCCGTTGCCCTCGGA CTTGCTTAGGGATAGATGGGAAGTGCCTATCCAAAGGAAGAGACCCAGATTGG TGGATGGGAATGAGGGGCGTGGCCTCCCGTAGACTCAGGGCTCAAGTTGGACG TGGGCCCAAATCTGGACCGGCTGGGTTTGCTGGGGGTGTCTTGAGGTCCCCTC CACCGTCGTCTCCGAATCCCCCTCCATGATCCTTCCTTGCTCCATCTCACCCTG GCAGAAAAAATCTAAGATCTCCGCCTCGAGAAAATTGCAGCTGAAGGTGAGGAC GGGCGGGACTGGGAAAGAGCAGGCAGGTGCTCAGGGGGCGGAGCTTGAGAATGGG TGGGGCTTTCGGGGTAGGTGGGCGGAAGTGGGCGGGGTTTGGCCGCTCGGGGCGTG GCTTTAGCGGGGAGTACTGCTCGGGGTGGGACGGGGCCTTGGAACAGTGGAGA CCAAACTGGAGGGTTTAGAAGGGCAGAGGCGGTTCCCCACGCCTGGTCTTTAT CCTGAAGCCCCGGGTGGGCTGCGCTTCCCTCCCACCCCTCTGCAGACTCTGCTG CTGCAGATTGCAAAGCAAGAGCTGGAGCGAGAGGCGGAGGAGCGGCGCGGAGAGAAGGGGCGCGCTCTGAGCACCCGCTGCCAGCCGCTGGAGTTGGCCGGGCTG GGCTTCGCGGAGCTGCAGGTACCGGCTCCCAAGGATGCGAGGTTTCTAGTCCC GGAATTCAGCAGTACAGCCTCTATCCCCTCTTCTGCTCGGGACCCAGGCGTCCA ATATGGCTGTCCTTACCCAATTATATATGGTTCGTGGGACTCCTGGCCCCTAACAC CCTTTGTGTGCAGGTCTGTGGAGTCTTGGCTCCAACCTACTCCTTCAGGACCATGTGG CCCTCCTATCCCAGACAGAAGCCCAAGCCCCAGCCCCTCCTCCCTCAGACCCTGGAG TCCAGGCCCCAGCCCCTCCTCCCTCAGACCCAGGAGTCCAGTCCCCAGCCCCTCCTC CCTGAGACCCTGAAGTCCAGGCCCCAGCCCCTCCTCCCTCAGACCCTGGAGTCCAGG CCCCAGCCCCTCCTCCCTCAGACCCTGGAGTCCAGGCCCCAGCCCCTCCTCCCTCAG ACCCTGGAGTCCAGGCCCCCAGCCCCTCCTCCCTCAGACCCTGGAGTCCAGGCCCCA GCCCCTCCTCCCTCAGACCCAGGAGTCCAGTCCCCAGCCCCTCCTCCCTCAGACCCTG GAATCGAGGCCCTCAGCCCCTCCTCCCTCAGACCCTGAAGTCCAGGCCCCAGCCCCT CCTCCCTCAGACCCTGAAGTCCAGGCCCCAGCCCCTCCTCCCTCAGACCCTGAAGTC CAGGCCCCAGCCCCTCCTCCCTCAGACCCAGGAGTCCAGTCCCCAGCCCCTCCTCCC TCAGACCCTGGAGTCCAGGCCCCAGCCCCTCCTCCCTCAGACCCTGGAGTCCAGGCC CCAGCCTCTCCTCCCTGAGACCCTGGAGTCCAGGCCCCAGCCTCTCCTCCCTCAGACC CTGGAGTCCAGGCCCCAGCCCCTCCTCCCTCAGACCCTGGAGTCCAGGCCCCAGCTC CTCCTCCCTCAGACCCTGGAGTCCAGGCCCCAGCCCCTCCTCCCTCAGACCCTGGAG TCCAGGCCCCAGCCCCTCCTCCCTCAGACCCAGGAGTCCAGTCCCCAGCCCCTCCTC CCTCAGACCCTGAAGTCCAGGCCCCAGCCCCTCCTCCCTCAGACCCAGGAGTCCAGT CCCCAGCCCCTCCTCCCTCAGACCCAGGAGTCCAGTCCCCAGCCCCTCCTCCCTCAG ACCCTGGAATCGAGGCCCTCAGCCCCTCCTCCCTCAGACCCAGGAGTCCAGGCCCCC AGCCCCTTCTCCTCCCTCAAACCCAGGAGTCCAGGCCCCCAACTCCTCCCTCAGACC CAGGAGTCCAGGCCCCCAGCCCCTTCTCCTCCCTCAGACCCAGGAGTCCAGGCCCCG AGCCCCTTCTCCCTCAGACCCAGGAGTCCAGGTCTCCCTGTTTTTGGTTCCCCCAACA ACACACACCACGTTCCTCCTCCAGGACTTGTGCCGACAGCTCCACGCCCGTGTGG ACAAGGTGGATGAAGAGAGATACGACATAGAGGCAAAAGTCACCAAGAACATC ACGGAGGTGGGACGCATGGGCAGCTCGGGTACCTTCGGGGTAGGGTGAGATGG CTGGGACTTGGTCTCTGCCTGACCCCTTGCAGCTGCTTTTGGCTGCACATCCCA GGAGACCCAGGACAACTGTGAGCCTGGCAGGGCTGGGGCAGAAGGATGAGTAC AATATAGTCAAGGAAAGCTGTTCTAGGCAGAGGGAACAGCACATGCAAGGCCA TGGGTTGGGAAACAGAAAATAAGTTAGTGAACATGCTCAGGGCATCACATGTTG GTAAATTAGCTCAGGCACTGGCCAGGGAATTGTGATTTGCATGTAGCTGGACCAGGTTATGCCAGTGGTTTTGAGAGGTGAGGCTGGAGCATATGAGGAGGGGGATT CAGTTCCAGGATTAGAAGCCTAGACTGGGAGCCTAAGCCGGGAAGAGACTGGT AAGGCCTCGGTACTGGAAGACGAGATAAGGAGAATAAAAAAGGAGTGTAGGAT GGAGGAGTTGGGTGTGCGGGAAATGGAAGGAGAAGTACCCACCCCCTCGTGTG CCCCCAGATTGCAGATCTGACTCAGAAGATCTTTGACCTTCGAGGCAAGTTTAA GCGGCCCACCCTGCGGAGAGTGAGGATCTCTGCAGATGCCATGATGCAGGCGC TGCTGGGGGCCCGGGCTAAGGAGTCCCTGGACCTGCGGGCCCACCTCAAGCAG GTGAAGAAGGAGGACACCGAGAAGGTGAGTGTGGGCTAAGGCCAGGAAAGAGGA TGCTGAGGGGAAGGGCTGTGGGTGCCAACAACCCTAGGCCTGAGGGCAGATGGTGC TTGGAGTTGGAGGTAGAAGCAGCTAGTAAGGGGTCCTCAGAAATGCAAGAGGAAGA CAGGAAGTAGAAGGGGAAGACAGGAAGTGCATTAGGGCTACAGGAAGTCCATGTAA GAGCAAAGAGGTACATGAAGGTCCAGGTGCAGTGGCTTATGCCTGTAATCTTTGAGG TTTAAAAAAAATTTTTTTTTAATTTCCTTTTTTTTTTTTTTTTGCGATGGAGTCTCTGCT GCCCAGGCTGGAGTGCAATGACACCATCTCGGTTGCTGGAGTGCAGTGACACCATCT CGGCTCACTGCAACCTCTGCCTCCCAGGATCAAGTGATTCTCATGCCTCAGCCTCCCG AGTAGCTGGGACTACAGGTGCACACCACCATGTTGTTGTTTTAATTTCTACAAATTTC TTTTTTAAATTAGCCAGTCACGGTGGTGGCATGCAGCTTCTCAGGAGGCTGAGGCAG GAAAATCGCTTGAGCCCATGAGATCGAGGGTGCAGTGAGCTATGACTGCACCACTGC ACTCCAGCCTGGGCAATACAGTGAGACCCTATCTTAAAGAAGAAGACAGGAAAGAT AGGAGTGGATGCCTGAAACCATGGATTGTACTGAACCCTATATGTAATTTTTTTTCCT ATACACACATACATACCTATGATAAAGTTTCATTTCTAAATTAGACACAGTAAGAGA TTAACAATAAATAACAACAAAAGAAGGTTGGGCGTGGTGGCTCATGCCTGTAATCTC AACACTTTGGGATGCTAAGGCAGGCAGATTGCTTGAGCTCTGGAGTTCGAGACCAGC CTGGGCAACATGGCGAGACCATGTCTCTATAAAAAATACCAAAGTTACCGGGTGTGG TGGCTCGTGCCTGTAATCCCAGCGCTTTGGGAGGCCAAGGCGGGCGGATCATGAGGT CAGGAGTTTGAGACCAGCCTGGCCAACACAGTGAAACCCCACCTGTAGTAAAAATA CAAACATTTGCTGGGCGTGGTCGTGGGTGCCTGTAATCCCAGCTACTTGTGAAGCTG AGGCAGGAGAATTGCTTGAACCCGGGAGGCAGAGGTTGCAGTGAGCCGAGATCTCA CCACTGCACTCCAGCCCAGGCAACACTGGGAGACTCCATCTCAAAACAAAACAAAA CAGAACAAAAGTTAGCTGGGCATGGTGGCACACACCTGTGGTCCCAGCTCCTCAGGA GTCTGAGGTGAGAGGATGGCTTGAGCCCAGGAAGTTGAGGCTGCAGTGAGCCGAGA TTGCACGACTGCACTCCAGCTTGGATGAGGCAGCCAGACCCTGTCTCAAATAATAAT AATAATAAAATAGAACAATTATAACAGATTGTAATAAAACTCATGAATGTGGTCTCTTTCTCAAAATATCTTATAGCACTGTAATCACCCTTCTTTTCCTTGTGATGTAAAATGA CAGTGCCTATGTGCTGAGATGAGGTGAGGTGGATGACATAGGCATTATGACCTGGCG TTAGGCTACTATTGACCTGAGAATCCATCAAACTTATGAATTGTTTATTTCTGGAATT TTCCATTTAATATTTTTGGGCCATGGTTTACCTCAGGTAACTGAAACCACACAAAGTA AAATTGCAGAAAAGGAGGGACTACTATAATAAGAAGAGAAGGAAGGAGACAGGAA GGGCATAAGGCAGACAGGAAGTGGAGGGGAAAGATAGGAAGTGCAGGAAGGAGAC AGGAAGTGGAGGGGAAAGACAGGAAGTGCATGAGGGAGACAGGAAGTGGATGGGG AAAGACAGAAAGTACATGAGGGAGACAGGAAGTGCTGGGGAAAGACAGGAAGTGC ATGAGGGGGACAGGAAGTGCATGAGGGAGACAGGAAGTGCATGGGGAAAATTGGC AGGGATTATCTTGAAAAGACAGGAAGTGCTCCAGAACTAGATACTTAGGCATCCAG GGTAGAGTGGCCCCACAGGCTGGAGGGAAGACAGGGATTCTTGAGAGACTGGAGAC CAAGAAGAGACCCTAACCTCTGACTCATCGCCATCCTCCAGGAAAACCGGGAGGTG GGAGACTGGCGCAAGAACATCGATGCACTGAGTGGAATGGAGGGCCGCAAGAA AAAGTTTGAGAGCTGAGCCTTCCTGCCTACTGCCCCTGCCCTGAGGAGGGCCCT GAGGAATAAAGCTTCTCTCTGAGCTGAAA Wild type HTT gene sequence with bold exons SEQ ID 223: HTT >chromosome:GRCh38:4:3041363:3243957:1 – indels are indicated by square brackets AGGGATGAGGAGCCCGCCCTCCTGGGCTTTAGTGGCGCTTAGGAGCCTGGGAGCT TCCCGTGGAGGTTAAGGGCGCTGAAACTGCACAAGCTGAGAGCTCACAGCAGTGC GTGGTGCCTCTTCAGCTGGGGTGCTCACCCGCGGGGAGACGTGGCTTTGTGCGCT TGGCGAACCCCACGCCAGGGCGTAGCGGAAGGGTGTGGACCGGCACTGAAGCCC GCGGAGGCACCCGTGCGTGTGCCAGGGCATGAAGATTTCCCAGGGGGCGACGTT GGAGCTGGGCGGGGAGCGAACAGGAGGGACAGGGAGGGCTGAGGCCCCAGAGC TGACCCTGCTGAGGGCCCTCGCTGGCGGCCTTGAAGCTGCGCCCAGATCCTTAGA GAAGGTGCGGAACTTGGGCCTTTTTCCTGAACTGTGCTTCGCAGGATGGGCCGGC CCGGCGCTGGCTGCGGCTTCCGCAGGGCAGGGCAGCACTTGACATGGTGAGGGC TGGTTCAGGGAGGAGCTGACGGGGACACCAGCCGGGTGCCTTCACCTCCGCCCCA TCCATTCTCCACTGCTGTCAGGGCTGCCTGGAGCCTGCAGCCGGGATCTCTGTGC TGCCTGCTGCGTGAGAAGCCAGGGCACAGGCCTCGGTGACTTTGAACCAGACAGT GACACATCGGACAGCTGCGCGCGGGAGAGGAACCACAGTGCCTGAGGGACACAG CGCTCCCTGGGCCGCTTTCACAAGAGCAGCAGGAGCGACCAGGCTTCCAGGCGGA CGTTTCCCTTGCATCGGGGTGTGGGGAAAATCAGCTTCCGCAGGTGCCCACCGCC GGCTCCAGCAGGGGCACTGGGGCCCGTCCTGGGAGGAAAACACCAGCGGGGATG TCACCATGAGGTTGGGAGCCTTCTTTGTACGAGGGGAAGGGGAAGGGGACGTTATCTT GGAGACTTGGGGTCTTTATTCTTTGGGGGAATGGAAATAGCTTTGCTTTTCTTCTGATTGAGGCAAAACTCATTAAAGAGTAAGGCTTTTTGGAAGCGGGTGACCGCGGCGGGACTCT GGACCCCTCTCCTCTGTGCCTCACTGTCCTCAGCTGCAGAATGAAGATGGCAGGAGTTG GTGCCCCCGGGATGCTGAGAGCAGGTTTCAGAGTCTGTAAATCATGTAGCACAGTGTTG AATACGTGGTCACCATTCAGTAACTATGCCAATAATCGTGGGAAAATAATACCTGCTCA TTGCAGAAAATACAGGAAAACATGCAGGAAAGAAAAATCACCATTATTCCACCCCCAC CCCCATTTCCACTCTGACTCTTTCCAGGAGTTCTTCCATCAGTGTAGACAAAGTCCACAG CGGAATCACAGTGTAGACGACTCTGCTGGGGGCTGGGGACCTGGCTGGGCTGAGGACC TGACTTCCTTGCCAGCCACAGGGCCTCTGTAAAGGCCTGCCTCGCGCAGGTGTTGAAGT TTAGGGACACCTGCCTGTCAGAGGCATTTGAACAAGAGCAACTCCATCTTGAATGGGG GCTGGGTAAAATGAGCCTGAGACCTGCTGGGCCGCATTCCCAGGAGGTTAGGCATTCTT AGTTACAGGATGAGATAGGAGGCTGGCACAAAATACAGGTCCCAAAGAGCTTGCTGAT AAAACAGACTGTTTTAAAGAAGCCGGAGCTAGGCGCGGTGGCTCATGCCTGTAATCCC AGCACTTTGGGAGGCCGAGGCAGGTGGATCACTTGAGGCCAGGAGTTAGAGACCACCC TGGCCAAAATAGTGAACCCCGTCTCTAATACAAAAATTAGCCAGGTGCAGTGATGGAT GCCTGTGGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATCTCTTGAACCCAGGAGT CAGAGGCTGCAGTGAGCCGAGACCTGGGCAACAGAGCAAGACATCGTCTCAAAAAAA ATAATAAAATAAAGAAGTAAATAAAGAAGCTGGGAAACCCACCAAAACCAAGATGGC GATGAGAGTGACCTCTGGTCATCCTCACAGCTCATTACATGCTAATTATAATGCATTAA CATGCCAAAAGACACTCCCATCAGCGCCATGACAGTTTTCAGATGCCATGGCAACGTCT GAAAGCTACCCTGTATGTTCTAAAAAGAGGAGGAACTCTCAGTTCTGGGAATCGCCCA CCCCTTTCCCAGAAATCTCATGAATAATCCACCCCTTGTTTAGCATATAATCAAGAAAT AACCATAAAAATGGGCAGCCAGCAGCCCTCTAGGCTGCTCTGCCTACGGAGTAGCCAT TCTTTATTCCTTTACTTTCCTAAGAAACTTGCTTTCCCTTTATGGATTAGCCTCGAATTGT TTCCTGAGCGAGATCCAAGTACCCTCTTTTGGGGTCTGGATCGGGACCCCTGTCCAGTA GCACTCCCTCCATCTTGCCTGCTGAGGAGGAAACTAGACCTGGGAACCGACCAGGGTT GCAGGGCCCAGGGCCCGAGGGCATCGTTCATACCCTGGGCTGGTGGCTATGGAACTAA TAAAGTCCTTTTTGTTTCTTTGTTTCTTTTTTTGCTTAAGCTAGTCTGAGTTAAATTTCCC CATGCCCAAGTGAAAGCTTTGACTGAGAGAAATAAACCTAGGCCAGGATTTGTACACA GACCATTTGGTAACTTGTTCAATTAACATGTTGAACATCTCATTTTTCTCTTTCTGTTTAT AAGCAAGCATGTATGTATTGAAGAGAACTTTGAAATTAAGAAGAACAGTGCTCGCTGT GGCAGCATATACACTAAAACTGGAATGATACAGAGAAGATTAGCATTGCCCCTGCACA AGGACGACACACAAATTTGTGAAGCGTTCCATACAAAAATATATTTAAAAAAAGAAAA AACACAAAATCTCGTCTTTAATATTAGCTGCTGTGCATAGGCAATCCGTATCAACATTT TGTTTGCTATTTATATAATTAATATAGTGATTTATTTCTTTCCATGTCATTTGGCTGCTTA AATTTTTAATGTTTTTTTTGAGAGTCTCGCTCTGTTGCCCAGGCTGGAGTGCGGTGGCGT GATCTCGGCTCACTGCAACCTGCATTCAAGTGATTCTCATGCCTCAGCTTCCCGGATAG CTGGGATTACAGGCACCACCACCACACCGGCTAATTTTTTTTTTTTAGTACAGATGGGG TTTCACCATGTTGACCAGGCTGGTCTCGAACCCCTGGCCTTAAGTAATCTGCCCATCTCA GCCTCCCAAAGGATTACAGGTGTGAGCCACTGTGCCTGGCCTGCTTAATCTTTTATTAC ATATTTCAGTCATCCATGATATTGTGAAGTTATATTTGGTTTTCGTTCAGTCTCCTGGCA TATATTTCCCCAAATCCTTGGAATCTTCAAAGTGATGAATGTCTTTTTTGTATGTTAATG AGGTGGCTGGTGGCTCCTAGACAGCTTCAAGATAGGAGGTGGGGCTGGTCACCAGAAA GAAGAAGGTAGGGTTAGAAGGTTGGGACTCAGCCCTACTCTTGCCCTCTGGGAAGGGA GAGGGGCTGATGGTTGAGCTGATCGCCAATGGCTAATGATTAAATCATGCCTATGTGAT GAAGCCTCCATAAAAACCCAAGAGGACAGGGTTCAGAGAGCTTCCGGGTAACGTGGAG GTTTCTAGAGGGCAGCGCACCTCCCTTCTACGGGGACAGAAGCTCCTGTGCTTGGGACC CTTCCAGAACGCTCCCTATGCATCTTCTCATCTGGCTGTTCATCTGTATCCTGTAAAATA TGCCTTGTAATAAACCAGGAAATGTGTTTCTCTGAGTTCTGTATGCTGATCTAGCAAATT AATTGAACCCAAGGAGGGGGCTGTGGGAACCCCCAATGTTTAGCAGGTCAGTCAGAATCACAGGCCAAATAACCAGGGGCTTGCAGTTGGCATTGGAAACGGGGTTCAGTCTTGTG GGAACTGAGTCCTCGATCTGTGTGGTCAGGCGCTGTCTACAGGTAGATGCCGTCAGACT TGAATGAGAGGACACCCAGCTGGTGTCCGCTGCAGAACTGATTGCTTGCTTGTGGGTGG GGAGAAGTATTCTGTGTTGATTCTTGAGTGAGAGAACAGGAAAAGGACACTGCGCTTTT CCCCTAGATCCTCAGGGTGGCTGAATATACATATGGAATACGCTGTAGGAGGAGTTATG AATATCTGTGAGGGGAGCAACACCCAGTGTGCAGTTAGCTTCATGCCTCTTCATGGGTC ACGTGTTCAAAAAATGTTGGGGTTAGCGTGATCGGAGGATGTGGTTTTCCTCCCTCTGA CGTCAAAAGGTGAAGCAGAGGACACGAGAACCCTCACTGCACCGCCTCTGGAGACGGC CACGGCCACTGCAGGCTCTGTGGTCTCTTACCAGGAAGGGAAGCATTGCCTGGCCGTGG CCTCAGATGATTGGCTAAAGGCAATAGAAGAGTGAGATGTCTGTTCTTTTTTTCTAGAG CTGGTTTCTGTTTATTCTTTAGGAAGGCATTCCAGTGAAAGGGTATTAAGGAAGGGGCA CACTGAGGCGAGTCTGACCTGTCCTGTCACGGCCAGGAACTCAGTTTTTAAGGTTTCTC TGAGGTCTCCTCGACCAAGAAGCGGTCCGCTCAGTTGGCAGGGGTTTTAGGATTTTCTT ATGTACCTCAAGGGAGCAGTAAACATCACATCATAATTTTTTTTTTAACAAAAAAGCAC AATATGCACTCAAAACAGTGTTTGAACATTTTAAAGAATACATATTATATACCTCTGTT AAATGAAAAACTTCAGCCAAATTAAATTTAAATGAGTTTAATTGAGCAATGAACGATTC ACGAATTGGGCAGCCCCCAGAATCACAGCAGATTCAGAGGGACTCCAGGGATACCTCG TGGTCAGAACAAATCTATAGACAAAAAAGGGAAGTGACATATAGAAATTGGAGGTGA GTACAGAAACAGCTGGGTTGGTTGCAGCTCGGCATTTGCCTTATTTGGACACAGTTTGA ACTCTCAGCAGTGTACGAGTGAAGTACGGCTGCTGGGATTGGCCAAGACTCAGGTACT GCCACAGGTGCAGACTCCCAAGTGAGGATGTCAGGGTTTCAGTCTTGTCTACCAATTAA GTTAGGTTGCAGTTCGTCCACAAGGACTCAAATAGAGAAGTATGGAGTCCTTCTCAGGC CATATTTAGTTTGCTTTAACAATTCCACCCTTTTGGTCATTTTCTCAATTTTGAGAGATTG ACCAAAACTTTAGTCATTGATGTCACTATCACCATCGTAAATGTACTTATTTGGTCCTGA AACCCTCTGAGAAACAGTAGAACAGTGAGTTTTGCAAAGGTAGGAACAAGGACTGAGG GTACCTCCTTATGCTGGAACGTCCTGTTTATAGGGGAATCTGTTAGGATCTATGTGTTTC CTTAAAGTCTTAGTTTGATTATGTCACATTTAGCATGAGTGACTCCATTTTTGTTTGGCT TAGTCTGTTGGGACATAGTGCATGAGCTCAGTCCAAAACAATGGCCTCCCATAATTTTA TTTTAATAAATTCCCCTTTCTGTCCAGGTTCTCACTTAGGTGAGTGTGGGCGGCAAGCCA CCCAGCTGCCAAGGCAAGACCCCGAGGGCACAAGCTGTTCCAGTATAATAAAAAATAT ATATATAGAATAAGAATAGTTATACTAGGCCGGGCGCAGTGGCTTACGCCTATAATCCC AGCACTTTGGGAGGCCGAGGTGGGCAGATCATGAGGTCAGGAGATCGAGACCATCCTG GCTAACACGGTGAAACCCCGTCTCTACTAAAAATACAAAAAATTAGCCGGGCGTGGTG GCGTGCACCTGTAGTCCCAGCTACTCGGGAGGCTGAGGCAGGAGAATGGCATGAACCC GGGAGGCGGAGCTTGCAGTGAGCCGAGATTGCGCCACTGCCCTCCAGCCTGGGTGACA GAGCGAGACTCTGTCTCAAAAAACAAAAACAAAAACAAAACAAAACAAAACAAAAAG AATAGTTATACTAGAAATAGATTATAGATATGATTAGATATGACTATTATCAATCATTA GTTTGTGGCATTACTCTATTCCAATATTATAATAATCTTTGTTCTACAATTATAACCTAG GAAAAACCAGGCCATACAGAGATAGGAGCTGAAGGGACAGGGTGAGAAGTGACCAGA AGACGAGTGTGAGCCCTCTGTCACGCTCAGACAGGGCCACTAGAGGGCTCCCTGGTCT AGTGGTAATGACAGTGCCTGGGAAGGCACCTGTTACTTAGCCGACCAGGGAAGGGAGT CTCCCTTTGGCCAGGGGAGAGGGGCGGAGGGGTTAGAGAAGACTCTGCTCCACCATCT CTTGTGGAAGGCCTGACATCAGTCAGGCCCGCCCACAGCCATCCGGAGGCCTGACCAT CTCCCTGTGATGCTGTGCTTCAGCGGTCACGCTCCTGTTTCACTTTCATGTTCCGCTCTG TACACCTGGCTCCATCTTCTAGATAGCAGTAGCAGAATTAGTGAAAGTGCTAAAGTCTT TGAAATGCATAGAAGAAATAATGACGTAGGCTGTCTCCTCTCTCTCTCCGCCTCGGCTA CCAAACAGGGAAGGGCACCCTGTCCGGTGGACACGTGACTTGCGTGACCTGACCTATC ATTGGAGATGACTCACACTCCTTACCCTGCCCCCTTGTCTTGTATCCAATAAGTAACAG GGCAGCCAGGCATTCGGGGCCACTACCAGTCTCCGTGTCTAGGTGGTAGTGGTCCCCTGGGCCCAGCTGTCTTTTCTTCTATCTCTTTGTCTTGTGTCCTTATTTCTGTGATCTCTCATC TCCGCAGGTGAGGAGAAAAACCCACAGGCCCAGTAGGGCTGGACCCTACAGGCGAGA GTGTGACCAAAACTTAGGGCCTTAGCACCACTCTCAGTTACCATCGTTTTGGGTTTCTG GTCTCAGCACGTCATTCATAGGTTACGGTGTCCTCATGGTCACACATTTCTTTCAGCTTC TGCCATTCCAGTTGAAGAGAGACCATTTGTTGTTCTAGAGATGGCTGCATGCAGACATT TAAAACCTTTGAGAGAATGCAGCGCACCAGGGAGACTATTATTATGATTACTGGGAGG ATAATACCAAGAGTTTGCAGTATGCTCCTTACCCAGGGTCCCTATACACCAAACCACCC CAAATCAAATAGATCAGACAATGAGCTAGATAAGAGGCTACTCACTCAACTAAGTGGC CTTTTCATTAATCTCCCACAACTGAATCTCTATAATACCTGATGTATTTCTCCCTAGGCC ACAGGTGCCAGCAGCTGCACAGGTACTACTTTTCTGTTTAGTCAATTCTATTATTTAGCA TACCTTTCACAAGGGAATTGAAAGTCTGTTGTGTAACTATAGCCTTTACAGTAGAGTCT GCTATACAGCCTATCATGAGGGATACATTTCTGGTTATGGCCTCTTTTACTCTAAACCAT GGAAAAGTGATGCCCTTCTGGAAGAGTAAAGCCCTCCTGGCAATGCCCTCTTTAACCCA TAGTGTGGGTTAAGAGGAGTGAACCGATATTCTGTTTCTGACTGATTACAGGCAACATA TGTCTCATTAAAGTTTCTCACCTACACTGGACCTTCATCTTTTATCTATCAAAGTATAAA GTTATCCGTGTATAAGGCTGGCTGCAAAATCCTTCACAAATGAAAGTATACCCCATAAG TGCACACAACAGATCGTCTTTTCATTTCTATTATTCATAGAGGCATAAGCAAGGAACAG ATACTCAAAGATAAGAGTCTCATGGCCAGGCGTGGTGGTGCATGCCTGTAATCCCAGC ACTTTGGGAGGCCGAGGTGGGCGGATCACCTGAGGTTAGAAGTTCAAAACCAGCCTGG CCAACTTGGCAAAAACCCATCTCTACTAAAAATACAAAAATTAGCTTGGTGTGGTGGCA GGCACCTGTAATACCAGCTACTCGGAAGGCTGAGGTATGAGAATCGCTTGAACCCAGG AGGCAGAGGTTGCAGTGAGCTGAGACTGCACCACCGCACTCCAGCCTGGGTGACAGAG CAAGACTCCATCTCCAAAAAAGAAAAAAAAAAGAGAGTCTCATGACAGTAGAAGTCTT GATCCACGATCTTAGCAAAAGCCGTTCACATCAAGGATGCCATCTTCTGGGGAGAGACT TCCTTGGCTTTACCTTAAAGGTTCCAATGGGTGTACAGTTCCAGGAGTGTGGATGGACC CTTCTCAGTTGTGAGATTATGAACCCAAGTTTCAAGATCCCTAAGTTTTGTTGCAGTGCG GATGGCAAGGACAGTCTTTCTCTGATGTTCTCAGAAGATCCAGTCTTCGGGTTCTAGAA TGCAAAGGGTTTGACCTCAGTGAACCATAGAAAAATATACTGTAGCATAATAACCTACT GTTAAAACATCAGCCCTCTTAACATGCGAGAGCTTTTATACAACCCGGAAAACATGCAT TGAAAATGACGATTGAATAAAATCCCTTTATAAAATATGTAAATAGCCCAGGGTTATAA ACTATTTTAGCAATTTGTAAGTTATCACACACACACACACACACACACACACACACACA CACACACATATACACTTTTTTTTTTTTGAGATGGAATCTCTGTCACCCAGGCTGGAGTGC AGCGGTGCAATCTCGGCTCACTCCAACCTCTGCCTCCCACACACCAATGTATTTAATTT GGATTACTTTATCTTTTCCATGATGAGTCACGAAATGCAGAACTTTTAATAACAAAAGC TTTAAGGACTCAGGAAGGACAAGGTGGCCGTCCTGGTTCTCCGTGAGTCCATACTTAAT TAACATTAGACTTCAGTCCTCTTGAATCCCAGTTGCTTCTCCAAATTAGGTGCATTGCAC GGATAACTGATGGGTTATCACAGGTAATTTGACTTAGACCATGGATTTCATTCAAATTG TATATCTGAACTATTTCAGTGTCAGCTGATTTAAGATGAAAATCTGGCAAACTCTTTTCT TGGTATTCAATTAATTTTTGTTCTACTTGGGTTAGCAGTTCTATAAGCCAGTCAGTCTTT TCATTAAAGTTCCAGGAATTCTTACCCAGTTCAAATGATATGATTCTAAAGTTATTAGA AACCTGTATTCCAAGACTGATTTTTAGGGTCCTTTTCATCCTTTTATGAATCTCCTAAAA GACACCATACTTTTGAAGTTTTCAGAAACCGCACCAGCATTAAGCAATTAACTGTGGAA ATGACTTTAAATAGTCATAGTTAAAAACACAATTGACAAGGAAATTTGGTTATTTCTAT GATTTACAGTAACTTAACCATAATTATATTGATAGCATAGACTCAGGCATATTAGAATT TTAGAAATCCCATTTAATTTTGGAACATATATTAATATCATTCACTAAAATGTAACCTG AAGAGGAGTAAACATTATTTTTTGACAATGCTTCCCATGTAACTTAACATATCAGATTA TCCCGTTTACCTCTCTTTGGATGCTTTAGGGGATCTCTGTAACACCCCAAAGTTAGAGGT CAGAAAGACTATTTTGAAGCTGAAATTTGATTTTGGGAAGCCTATCAAATATATTAAAG GTTTAAAACACTTGATGTTATGCTTAACCAGTTTGACCATGAGGTGAGATTCTTATAAACCTTTTATAATCCCTTACAATTTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTG TGTGTGTGTGTGTTTTAGATGGAGTTTCATTCTTGTTGCCCAGGCTGGAGTACAGTGACA GGATCTCGGCTACCGCAACCTCTGCCTCCCAGGTTCAAGTAATTCTCCTGCCTCAGCCTC TCTAGTAGCTGGGATTACAGGCATGTGCCACCATGCCCAGCTAATTTTGTATTTTTAGTA GAGACGGGGTTTCTCCACGTTGGTCAGGCTGGTCTCGAACTCCCGACCTCAGGTGATCC ACCTGCCTTGGCCCCCCAAAGTGCTGGGATTATAGGTGTGAGCCACCGCGCCTGGCCTT TTGCCAGTTTGATATTTGGTGCCCCCATGCAGCCAGTTGGGTGGCAACTTGCTGCTCAA ATTTAAAAGTTGAGAGGCTTTTGCCTGTGGTTCAATGAAACAAAAAAAGATCATTTTCC TTTATGATGTGGCTTGGCCCTCAGGGATATGATGTGGAGAGCTGGGTCACTAGGGCTAC TCAGGGAAAGGGAATCCAGAAGCCTGGCACACAGCAAAAGGGTAAGAATTTCTTACCA GTCAGATTTCTGACCTCTCTCTCTCTGTGTGTTCAAACTGGTTGAATGAATTAAAAAAA AAAAAATCACTGCTTATCTCCTCTGTAAAGTTTTGATTAATGGAAAAAAGGATTTGTGA GGCTAATCTCAGGCTGTAGTGAATCTCGTAGGCTTTATGTGTCTTCCCGTGCTGTTCTGT GATAAAAAGGGGCGCCTTAGGATAGAACATGGGCTTAAAACCCCATAAGCTTGCTCGC CGCTCAAGATGGCCCAACAAGCTGGTCAGTAACTGCTGCAGATCCCTGAAACAAACAA AAAAACTAGATGAGGTCTCCATCTTGTTTTAAGTCTTTGGGAGTTTGACCTTGTAACCAT GTGGCGGTTACTTGGTCTCCACCTTCCAAGTGGAACAGGAATTTTGGGTTCATGTCATA GTTAGCTCTACAATCTATCTTGAGTAGTTACGAGGCTTTGCCAGCTGAAAATTAACTAC TCTAGACTCTTTCTGGGAAGGGCAATGGAGACTGCCCAGTGCTATAGTTCAGTGGTGAT CTGGGTTTGATTCCTGGCTTAGGGAATGAGTACTTTCTAATTGATATTTGGGTAACCTTT GCCATTTTTTGATTTTCTCCCCCTCCACAAACTGTCTTGAATTGCCTTTCTCAGAGCACC TGGGAGGTTACTTTCAATAGTTTAAAGGCAGGAATATTGGCTGTTTGGCCTGATGAAAG TTGGGTAATAAGCAATTTAAAAGAACTTTTATTAAAGAGTGCAATGGTTAAAAGTCAGC TTAATTAAAAGTAGATATTCTGGCTCTAACAGCCTGGGACTCCTTGGGAAAACAGGAG GTGCCAGAGACCCCGTTTTGGGAAAAAACTCTGTTTTACTCATGAAACCCCAGAAATTG GAAGTAGATAGATTCCTTTCAAAATCTAAGGCTCTGTTCTGTTTTGCATTGGATTATCTA CTGTTTTTTACTTGGGGGTATCAGAAATTACTTCACATAGAGACTTGGTGTGTAATAACT AGGCAGGAAATATACTTTAGGGATGTCTAATGGCAGTTATGGGGCTGACTCCCTCTTTT TTGGGGGGGATCCAGGATCTGGTATAAAAACGAGACCTCAGGCTGGGCATGGTGGCTC ACGCCTGTAATCCTAGCACTTTGAGAGGCCGAGGTGGGTGGATTGCCTGAGCTCAGGA GTTCGAGACCAGCCTGGGCAACATGGTGAAACCCCACCTCTACTAAAATAAAAATAAA AAAAAATTAGCCGGGCATGGCGGCATGCGCCTATAGTCCCAGCTATTTGGGAGGCTGA GGCAGAAGAATCGCTTTAACCCAGGAGGCGGAGGTTGCAGTGAACTGAGATGGTGCCA CTGCACTCCAGCCTGGTGACAGAGCAAGACTCCGTCTCAAAAAAAAAAAAGAGACCCT TAATTTTTGGAGATCCGTTTTGCCTTCCAGCTGTGCCTGCTTATTATATTAGGCCCTAGA AACTGCATGCTTTCCTGGTCCTGTTCTTCCAAGGACTCCATCCTAAAGCCAGTAATCCA ATTTAGAAACTTAGAAAGTGGTAAATGAAAAATCTTACAACTATTGGATCTTCTTCTGT CTGTCTGTGTAGTTATATATGTGTTGTGTGTTTAATGTTTATATAAAAGAGCTCTAATTG CCAGGCACATTGGCTCACGCCTATAATCCCAACACTTTGGGAGGCTGAGGTGGGTGGAT CGCCTGAGGTCAGGAGTTCGAGACCAGCCTGGCCAACCAGGCGAAACCCCGTCTCCAC TAAAAATACAAAAATTAGCCAGGTGTGGTGGCGCATGCTGGTAATCCCAGCTACTCAG GAGGCTGAGGCAGGAGAATCACTTGAACTCAGGAGGTGGAGGTTGCCCTGAGCCAAGA TTGTGCCACTGCACCCAAGTCTGGATGACAGAGTGAGACTCTGTCTCAAAATAAAAATA AAAATAAAAATAAATAAAAGAGCTCTAATTAATTGGCTTTAAGAAAAATGAGTGCTTA AATCAAATATTTTGAAAGAAAAATAAAAACTCTAATGCCTTTTAGTGCATGTAACTTCA GTAATCTTTGGGAAATAAAAAGTTTTAAAGGTTACTGGTGAAAATAAAGACATTTGGCC TAAATTAGGCAGGTTAGATATTAGGTTTGCTAACTTCTTTTTTTTTTTTTTTTTTGAGATG GAGTTTCACTCTTGTTGCCCAGGCTGGAGTCCAATGGCGTGATCTAGGCTCACTACAAC GTCTGCCTCTCGGGGTTCAAGCAATTCTCCTGCCTCAGCCTCCCGAGTAGCTTGGATTATAGGCATATGCCACCATGCCCAGCTAATTTTTGTATTTTTAGTAGAGATGAGGTTTCACC ATGTTGGCCAGGCTGGTCTTGAACTCCTGACCTCATGATCCCCCCGCCTCCGCCTCCCA AAGTGCTGGGATTACAGGTGTGAGCCACTGCACCCAGCCAGGTTTGCTAAATTTTTTAA GGTCATAAACTGCTTTGACTTTTCTAAATTGTTCAAATTATTTTGGAGCATTAGATTCTA GATAAGGCCTGCGGACATGTGGAATTAGCCATGCCCCCAGCTAGGCAAAGAAGATTAC AAAGAAAATAAATTTTATATAAGAAAGGATCTCGTATGGTAAATTCTTGCCCTAAAGTA AAATAACTGGTTGTTTAAAAAGAGGGGTGTTTAAAAAGAGGAGTGTTTAGGACAAGTG AGAAAGTCCAACCATGCCATAGATGGCCTGTGTAAGTTGTTAAAGGATTTGTGAAATTC ATGCACCAAAAGTAAAAGATGCTAAGAGTTACCATTATAACATGTAATTGAAACTACT AAAAAAATAGTTTTACATGCAAGGTGTGTGAGGAGAGTGAAATGTGTTTTTGGTAAAA GATTATAAGAAGGCATGGGAATGTAAATTTTTGCCTAGTTTAGAGGGTTAAAGGGTTTT TAAGTTAGATAAGATAAAGCTAAAAGTTTGAGCAAATTGTAGGTTTGTAAAAATTAATC TTGTAAAAGAAATTCTGTGTGTGAACATATTGACTAAATTTAAAGGAGTATTATTCATT TTTTCCAGAAATTGAACATTGAAGTAAAAGCACAACAGGGTTTTCTTTTCTTTCTATTTT TTTTTTTTGAGATGGAGTCTCGCTCTGTCACCCAGGCTGGAGTGCAGTGGTGCACTCTTG GCTCACTGCAACCTCTGCCCCCTGGGTTTAAGCGATTCTCCTGCCTCAGCCTCTGGAGTA GCTGTGATTATAGGCACTTGCCAGCACGCCTGGCTAATGTTTGTATTTTTAGTAGAGAC GGGGTTTCACCATATTGGCCAGGCTGGTCTTGAACTCCTGACCTCGTGATCTGCCCGCC TCCACCTCCCAAAGTGCTAGGATTACAGGCATGAGTCACCACGCCTGGCCAACAGGGTT TTCTTAAAGCACTGTTCTGCTCTTTCACAAAAATTGTAAGGGCTTGGCCAGGTGTGGTG GCTCACACCTGTAATCCCATCACTTTGGGAGGCTGAGGCAGGTAGATCACGAGGTCAG GAGATCGAGACCATCCGGGCGTGGTGGCGGGCGCCTGTAGTCCCAGCTACTCAGGAGG CTGAGGCAGTAGAATGGCATGAACCCAGGAGGCAGAGCTTGCAGTGAGCTGTGATGGC GCCACTGCACTCCAGCCTGGGCAACACAGCGAGACTCCGTCTAAAAAAAAAAAAAGTC TCAAACTAGCCCTGGGTTGGGCCCTGTCATCTTTAACCCATTTTTAACCCAGAGGGACT TTACTGAGGGGAGGGCCTCTAACCCAATCCCATGCTTTACTTAGGTAAAATGTACCCCA TTACTTATTCAAAGTCAGCCAATTTGTGCTGCAGCCTATCTCCTTTGGATTGCGATAGTA ACTAAGCTAAAAGGTTAGCAGATTTGATTTTTGGGAGCCTTCATTTTTAAATGCATGTC AGTGCACTGTTGTTCATTCAGAATGTTCCACTGTAGTCATCTTTAGTAAGATTTCACCAT TTCTGTAAGACTTCACCACTTTCCATGCCTAACGTGTAAGCCAGAAGGAACCCAGTTTT CCGGAAATTAAGGAACCCATTTTTACCTGAATATTGGCTTTACGCTCAGCTTCCCTTGAT TAATTTAGCCAATGATTTTTTCCTACCTAAGTGTGCAAGAAAAATGAAACAAAGGGGTA GAACACAAACATCCCCACGAATTTTCAAAAGCCAAATTTTACACCCCCTGCAATATTAC CATTTTCTACCAGTTTCTTTCAGACCCAGTCAGATGTAAGAGGCCTCTAACTGGATCCA GGCCGGTTAATTACCGTATCAAATCTATTCCTGGACCCATCCATTTTCTTTGGCGACTTC CAAACCCAGTTTGGATCAGAAATTTGCACAAAGAAAACTCAGAGAGCTCGAAACACAA ACTGTGGATCTCCAAAATCCAAGAGAGGACTTATCCATGATCCCCAGCTGCTCTGAGAA ATCAATGGACACAAGTGTGTCCAGCAGGTACCTTACTTGTTCACTCAGTGCCCCAGGGG TCATTAGAAGCTTTACTTTGGATCCCACTTCTGACACCATGTTAAAAGAAAAACTTCAG CCGAATTAAATTTAAAGGAGTTTAATTAAGCAATGAATGATTCGTGAATTGGGCAGTCC TCAGAATCCCAGCAGATTCAGAGAGACTCCAGGGATGCTTCATGGTCAGAACAAATTT ACAGACAAAAAACGGGAAGTGACGTACAGAAATTGGAGGTGAGGTACAGAAATAGCT GGTTGGTTGCAGCTCGGCATTTGCCTCATTTGAACTCTCAGCAGTGTATGAGTGAAATA TGGCTGCTGGGATTGGCAAAGACTCAGCTACTGTTACAGGTGCGTAAGTTACAGTTTCA ATCTTGTCTACCTATAAGTTAGGTTGTAGTTCATCCACAAAGACTCAAATACAGAAGTA CAGAGTCCTTCTCAGGCCATATTTAGTTTGCTTTTACACCTATATGACCAAACTGCCCTT CTGAAAATTTAGAACAATTTCCCCTTTCACCAAAAACGTATTATAATGCCCTGTTGCAG TTTTCACCAAAAATCAGGTTTTTCTTTTGGCAAACAATTTCACCAGAAACTAGGTATTTC CTTTTCTTTTCTTTCTTTTAAAATTTGTAGTAAAATACACATAAAATTTACCATCTTAATGATTTTTAAGCATAGGGTTCAGAAACAATGTCATACAGTCACCCCCACCATCTATCATTC ATTCAATCAGTTTGCACAGTTAACTCTTTGCATCTTGTAAAGCTGAAGCTCTGTCCCCAT GAAACACTAATTCCACCAGCCCCTGGCAATTACCATTCTGCCCTCTGTTTTTATGAATTT GACTACTCTACATCTCCTGTCTCAGTGGAATCAGACAGCATTTGCCCCTTGTGACTGGCT TTTTCTTTTTCTTTTTTTTTTGAGGCAGAGTCTCGCTCTGTCGCCCAGGCTGGAGTGCAGT GGCACAATCTCGGCTCACTGCAACCTCTGCCTCCCGGGTTCAAGCAATACTCCTGTCTC AGCCTCCCGAGTAGCTGGGACTACAGGCGCCTGCCACCAAGCCCAGCTAATTTTTTGTA TCTTTAGTAGAGACGGGGTTTCACCGTGTTGGCCAGGATGGTCTTCATCTCCTGACCTC GTGATCTGCCCGCCTCGGCCTCCCAAAGTGCTGGGATTACAGGCGTGAGCCACTGCACC CCACCCTGTGACTGGCTTTTTCACTTAGCATAATGACCTCAAGGTTCATCCATGTTGTAG TGTGTGTCAGAATTTCCCTCCTATTTAAGGCCAAAACTATTCCATTGTACATATGTTACT GGAAAGGGGTCCCAATGCTGACCCCAAGAGAGGGTTCTTGGATCTTGCTCAAGAAAGA ATTCAGACGAGTCCACAGAATAAAATGAAAGTAAGTTTACTAAGAAAGCCAAGGAATA GAGAATGGCTACTCCATGGGCAGAGCAGCCCCGATGGCCGCTGGTTGGCTATTTTTATG GTTATTTCTTTTTATTTATTTATTTATTTTTGAGACAGAGTCTCCCTTTGTTGCCCAGGCT GGAGTGCAGTGGCGATCTCGGCTCACTGCAAGCTCCGCCTCCCGGGTTCATGCCATTCT TCTGCCTCAGCCTCCTGAGTAGCTGGGACTATAGGCGCCCGCCTGTAATTTTTTGTATTT TTAGTAGAGACGGGGTTTCACCATGTTAGCCAGGATGGTCTCAATCTCCTGACCTTGTG ATCCGCCCGCCTCAGCCTCCCAAAGTGCTGGGATTACAGGCGTGAGCCACCACGCCCTG CTGATTATTTCTTGATTATATGCTAAACAAGAGGTGGATTATTCATGAGTTTTCCGTGAA AGGGGTGGGCAATTCCCAGAACTGAGGGTTTCTCCCCTTTTTAGAACATATAGGGTAAC TTCTAGGCCAGGCATGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCAAGGCG GGCAGATGACCTGAGGTCAGGAGTTTGAGACCAGCTTGACCAACATGGAGAAACCCCA TCTCTACTAAAAATAAAAAATTAGCTGGGCGTGGTGGCGCGTGCCTGTACTCCCAGCTA CTCGGGAGGCTGAGGCAGGAGAATCGCTTGAACCCGGGAAGCCGAGGTTGTGGTGAGC CGAGATTGCGCCATTGCACTCCAGCCTGCACAACAAGAATGAAACTCTGTCTCAAAAG AAAAAAAAAAAATAGAACATATAGGGTAACTTCTATATGTTGCCATGGCATCAGTAAA CTGTCATGGGGCTGGTGGGAGTGCCTTTTAGCATGTTAATGCATTATAATTAGCTTATA ATGAGCTGTGAGAACAACCAGAGATCCCTTTCATTGCCATCTTGGTTTTGGTGGGCTTT GGCCAGCTTCTTTACTGCAACCTGTTTTATCAGTAAGGTCTTTGTGACCTGTATCTTGTG TCAGCCTCCTACCTCATCCTGTGACTATGAATGCCTAACCTCCTGGGAACGCGGCCCAG CAGGTGTCAGCCTCATTTTACCCCGCCCCTATTCAAGATGAAGTTGTTCTGGTTCCAACG CCTCTGACATATTAGCTGCATCATTTTACATTTCTTTTTTTTTTTTCCTTTTAAATGGGGT CTTGCTCTGTCACCCAGGCTGGAGTGCTGTGGTATGATCTCGGCTCACTGCAATCTCCA CCTCCGAGGTTCCAGCGATTCTCTTGCCTCAGCCTCCCGAGTAGCTGGGACTACAGGCA CCCACCATCATACTGGGCTAATTTTTGTGTTTTTAGTAGAGATGGGGTTTCCCCATGTTG CCCAGGCTGATCTCAAACTCCTGGGCTTAAGCAATACAGCCGCGTTGGCCTCCCAAAGT GTTGGGATTACAAGCATGAGCTACCCCACCCAGCTCATTTTACATTTCCACTTGTTAAA CTGAAAACTGGCCCGAGAAAGCTTCTGTACTGCCATCCTTGCGTCCTTGCAGATGAATC GTAACCTAGCATAGTAGGTAGGCAGACTGAAAACCTAACTTAGCAGTAGGCTTCTGTA ACAACAGCTGTGTCTCAGCCAGTTCCTGCAGCCAGACTTCAACCACTCACAGGCCGCAA ACTGTTCAAACTGTGTTCGGAGAAGGCGAATTCATCTGGCTGTTAACGTGCCTCACTTC TGCTTTCTGTGGCCACTTTCCCTTTTCTGTCCATAAATTTGCTTTGACCACACAGCATCC CTAGAGTCTCCCTGAATCTGCTGTGATTCTGGGACCTGCACCATTTGTGAATTGTTTTTT TTTTCCTTGATCAGCTAAACTCTGTTCAATTCAATTTGTTGGAAGTTTTTAACATACCAA TGGTGCACCAAGGTTCCAATTTCTCCACTTCCTCATAAATAAGTCATTTTAAATGGCTTT TCAGTATTCCAATATTTGGAAGTATTAATGTTTCTACCAATTTTCTATTTTTGGACATTG AGGTTGTTTCATTTTTTTTTTCTTTTTTTGAGACAGAGTCTCGCTCCGTCACCCAGGCTGG AGTGCAGTGGCCTGATCCCGGCCCACTGCAACCTCCACCTCCCTCCTCAGCCTCCTGAGTAGCTGGGATTACAGGTGCATGCACCACCACACCCAGCTAATTTTTGTATTTTTAGTAG AGATGGGGTTTCACCATGTTGGTCAGGCTGGTCTCAAACTCCTGACCTCAGGTGGTCCA CCTGCCTTGGCCTCCCAAAATGCTGGGATTACAGGCCTGAGCCACTGCGCCTGGCCTCA TCTTCTTGATATTAATGTTGCTTTAACATCTTTGTCCCTGTGTTTTTTGTTTTTTTTTTTGA GACGGAGTCTCATTCATTCTGTCACCCAGGCTGGAGTTCAGTGGCGTGATCTCAGCTCA CTGCAACCTCTGTCTCCTGGGTTCCAGTGATTCTCCTGCGTCGGTCTCCTGAGTAGCTGT GTTCCTGGGTCTTTCGATGGTTATTTAATACTTCCCTACAGTAATGCCCTGTGCGTACAT GCTAAGTGTGATGAAATGGTTGGCACAGTTAAATCTTTTGAAAGACATTGCCAAGTCAC TCTTCAGAAAAGTGATAGGAGGTCATAGCAATTTTAAGAAGTCCTCATTTCTACATTTC CTTACTAATCTCGGTTGGTGTCTCTTCAATCTTTCCTCACACTTTTCTTGGGTTTTTCCTG AATCATGAGTCTACTACATTTACACATTTTAAAGCATCTTTAGAAACAGGATCTCATTTT GTTGCCCAGGCTAGAGTTTGGTGGCATGATTATAGCTCCTCATACTCCTGGGCTCAAGT GATCCTTCCACCTCTGAAACCCCAAAATTTGAGAAAGGTCTCATTTAATTTAGAAAGTT TATTTTGCCAAGGTTGAGGGTGCACACCTGTGATGATATACGAGTTAAAAAGAAATTAT TTAGGCAGATACTGAGGGTAAGAAAGTCCTCGGTAAGGTTTTCTTTTCAATGAAAAGCA GCCCCCAAGCATTTTCTTTTCTAACAAAGAGCAGCCTGTAAAATCGAGCTGCAGACATA CACAAGCAAGCTGGAAGCTTGCACAGGTGAATGCTGGCAGCTGTGCCAATAAGAAAAG GCTACCTGGGGCCAGGCAGATCCAACATGGCGGCTCCATCTTCCCTTTCCTTGTCAACC ATGTGCACAGTAAGGAGCAGGCAACATAGTGTCCCCCGAGTAGAGACCAATTTGCATA ATAAAAGGTGAGGGTAGGGTGGGCAGCTTCTTTGCATGCTATGTAAACATTATGCCTGG TCCAACCAATCTTTGGGCCCTGTGTAAATTAGACACCACCTCCTCAAGCCTGTCTATAA AACCCTGTCCATTCTGCCGCAGGCTGGAAGACCCACTGGGGCACCCCTCTCTCTCTATA GGAGACAGCTATTCATTTTTCTCTTTCTTTCACCTATTAAAGCTCCACTCTTAACCCCAC TCCGTGTGTATCTATGTTCTTGATTTCCTTGGCATGAGGCAATGAACCTTGGGTATTACC CCAGAACCTTGGGTATTATGCCACTTCAGTGACACAGCCTCAGGAAATCCTGATGACAT GTTCCCAAGATGGTCGGGGCACAGCTTGGTTTTATACATTTTAGGGAGACATGAGACGT CAATTCATATATGTAAGAAGTACATTGGTTCCGTCCAGAAAGGCGGGGACAACTTGAG GCAGGGAGAGAGCTTCTAGGTCACAGGTAGACAAATGGTTGCATTCTTTTGAATCTCCG ATAAGCCTTTCCAAAGGAGGCAATCAGAATATGCGTCTATTGACTGGGCGCAGTGGCTC ATGCCTGTAATGCCAGCACTTTGGGAGGCGGAGGTGGGTGGATCACCTGAGGTCAGGA GTTTGAGAGCAGCCCGGCCAACATGGTGAAACCCTGTCTCTACTAAAAATACAAAAAA TTAGCTGGGCGTGGTGGCGGGCGCCTGTAATCCCAGCTACTCGGGAGGCTGAGGCAGG AGAATAGCTTGAACCCAGAAGGAAGAGGTTGCAGTGAGCTGAGATGGTGCCATTGCAC TCCAGCCTGGGCAACAAGAGTGAAACTCCATCTCAGAAAAAAAAAAAAAAGGCCTGG GCAAAGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAAGCCGAGGCGGGCAGGTCAC AAAGTCAGGAGATTGAGACCATCCTGGCTAACATGATGAAACCCCATCTCTACTAAAA AATACAAAAAACTAGCTGGGTGTGGTGGCGAGCACCTGTAGTCCCAGCTACTCGGCAG GCTGAGGCAGGAGAATGGCGTGAACCGGGGAGGCGGAGCTTGCAGTGAGCCGAGATC ACACCACTGCACTCCAGCCCGGACGACAGGGCAAGACTCTATCTCAAATTAAAAAAAA AAAAAAAAAAAAAAAAAAAGAGAGAGAGAATATGCATCTATCTCAGTGAGCAGAAGG ATGACTTTGAATGGAATGGGAGCAGTTCCTAGCTTGAACTTCCCCTTTAGCTTCAGTGA TTTGGGGGCTCAAGGTATGTTCCTTTCACATACCTCAGCCTCCCAAGTAGCTGGGACCA CAAGTGCATGCCACCACACGTGGCTAATGTTTTATTTTTTTTGTAGGAATAGGGTCTCAC TATGTGTCCAGGCTGGTCTAAAACCCCTGAGCTCAAATGGTCCTCCCGCCTCAGCCTCC CGAAATGCTGGGATTACAGGCATGAGCCAGCATGCCCGGCCTAGTCTACATTTTTATAA ATTGCTAATTCAAAGTTCCCTCTCCAAAACCTCATGGTTTTCCCTGTTCTCATCCCCTGC ACCCTCCCTTCCCCTGGAGTACTCACCTGGCCTTGGAGGTCTGGTGTGAGCCCGGACTT CGATTCTAGGCACAGCATGTGATGAGCGCCCCCAGGTCAAACACCTCCCCTCTGCG GCCTGTGCTTCACCGCCTTGACAGTGAGAAAGGTCTCCCTTCGGCTCATTCTCGAAGTCTCAAACTTCACTTCTCCTGTGCGCTGATTCTGAATTCAGCCCCCGTCCAAGGT CCTGGCCCCTTTCTCTTCTGCTTGGCGTGTTGTTCATCACCACTGTGCACTGCTGA GGGTAAGTGCGGTTCTCTGGACCTCTGCTTTATCATTAGAACAGACTCTTGCGGTTTCC CACGACATTCCTTTCACTTCTCACTTGGAAGATGAGCCGTGAGGAAATCCTGTGTTGTG TGGTATGTGGGCTGTGCTTCTGCTTGACTTGAGGGCCAAGCAGCATTGCAAGCCATGGT TTTAAATAAGAAAGAACATTTCTAACCTTCATCTTCTAGTAAGGAAACAAGTGGGCTTT AGAGTTCTTGCTCAGGAAAGACCTATGTCCCAGTCCAACCGGACCTTTTACTAAAGAGA TCTTCCTGATCCTCCTCCCCAGGCCAGGGGAGGGGTCCTCCCTGGGGTTGGAGCCTTTA GTAGGGGGTCGGAGACACGACGTAGCCTTCATGACATTCATAGTCTAGTTACACGATCC CTGTAAGGGTCAGTTGAAGTAAGTGCTACAAAGGAAGGGAGGTGCTCAGTGGAGAGGG CTCTCTTTTATGTATTATATTTCTTTCATGGGGAGGGATATGGATCAGGGATCAGCAGA GGTGTTTCAGTCCCGAGGGAAAGAAAGTCAGCGTGGCTTGGGAGTTGGGAGCAGCAAG ACAGTGGCTCAAGATATCTTAAGACTAGTGGAGTACACCTTGCATGTTAAAAGCCTTGC TCAGGGCTGCCTGGTTCTTGTAGGACGACAGAGATGGCCTAGCTCTGCATACTGCACCC CCAGGGGCTCAGAACAGTGCAAATGTCAGTCTATCTGTCAGTGGCAGAGCCAGCCTTG GAGCAGGGGTGCAAGGAGGTCTCTGCACTGGCCAGGCATGCAGAACATTCTGTTCAGT AGCACTGGACAGAAGGCCCCATCTAGATGAGACAGAGCTGGTGGGGCAGGACAAAGA CTCCTGGCAGCTCAAACGGCCTGGCAGATGCTTGGAGAGAGGGGGCTTCTTGAGACAG CACCATTTCTGGGAAGAGAGTCACCTGGGAGGGATGAGGCCACGCTCCGGCTTGGAGG TGAAGAGAGGGGCTGCTGCAAGAAAGAATTAGAGACATGCCAGCCTTTGCTGTGTTGC CCAGGCTGGTCATGAACTCTTGGCCTCAAGCAATCTTCCCACCTCAGCCTCCCCAAGCG CTGGGATTATAGACATGAGCCCCCATGCTGGCCAATAAAAGATGATTTTATGGAGGGG ATGGTGGTGAAGGTTGTGGGTGGTATGAAATAGTAAGAAATATATATTGGTCTGCACCC AGTTCCTGCCACAGAGCTCCTAAAATCCTGAGAACTTCCTGGGTGAGCATCTTTTGTTCT AATGAGGTGACTCTTGGTGGCTCCTGGATAGGAGTGAATCACCAGAAAGATCAAGCCA GAGTTAGAAGCAGAAAGTGCTGGCTATAACACAGGAAAGCTGTAACACAAATAATAAA GTTTTTTTTTTTTTTTTTGAGATGGAGCCTCACTCTGTTGCCCAGGCTGGAGTGCAATGG TGCAATCTCAGCTCACTACAAGCTCTGCCTCCCAGGTTCAAGTGATTCTCCTGCCTCAGC CTCCTGAGCAGTTGGGACTACAGGTGTGTGCCACCACATCTGGCTAATTTTTGTATTTTT AGCAGAGACGGGGTTTCACCATATTAACCAGGCTGGCCTCAAACTCCTTACCTTGTGAT CCGCCTGCCTCAGCCTCCCAAAGTGCTGGGATTACAGGCATGAGCCACCGTGCCTGGCC AAAAGACATTGTTCTTAAAAGAATCAACTAACTAACCAAATAAATAAAAATCTAACCT AATTAAGAAACTAAAAATACACAAAAATTAATTTCAAGGGGAGAAAAATCATGTAAAG AGAGAAAGATAATGAATACTTTGCAGAAATTTATGAACATAAACATAAAACTTGGATG AAATGCATTTCTAGGAAAACATAATTTATCAAAACTAACCACAAGTAAAATAGAAGCC TAAATAGGATATTTTCAAGAGAAGAAGTAAAGTTGTCAAAGTGCTACCCTTCAAAAAA ACACCAGGCTCAAACAATCTGACATGGGAATGTTAGCACACCTTAGAGAGCAAATAAA ACTTTGAATGGGCTTGAAATATTCCAGACTCTAGAAAAACAAAACTTCCCAATTCTTTT TATAAAGCAAGTATAAATTGATACCAAAATCTTATAAAGACCTTATACAAAACTTCATA CCAATCTCTTTTATG[AATA / AATATTAATA]CAAAACCCTTAATAAAGTATTACCAGACA GAACCCAACAATACATAAAAATGTCACATCATAACATAGTGGGGTTTATTTCAATAATG CATGGATGGTTCAATACAAGGAAATTCAGTAACACAATATAATAGATCATGTGAATAT ACCCAAAGAAAAAATAGATTATTTTCATAGATGCTGTAAAGGCATTTGACCAAATTCAA CACCTACTTTTTAGGTGGTCAATAAAATAAATTAGTTACTCCTTCTTTAGCATGATAAAA TATATTTATCAGCCCAGAAGGCATCATTTTACCCGATAAGGGCACACGCTGGAGGGAAT AATGTTAAAATTAGGAATAAGAGGATAGCTAGTTTCTTTCTTCTTTTTTTTTTTTGAGAC GGAGTCTTGCTCTGTTGCCAGGCTGGAGTGCAGTGGTGCAATGTTGGCTCACTGCACGC CCCCCGCCTCCCAGGTTCAAGCGATTCTCCTGCCTCAGCCTCCCGAGTAGCTGGGACTA CAGGCGCGCACCACCATGCCCGGCTAATTTTTTTTTGTATTTTAGTAGAGATGGGGTTTCACCATGTTGGTCAGGCTGGTCTTGAACTCCCAACCTCACGTACTGGGATTACCGGTGTG AGCCACCACGCCAGCCCAACTACTTTCAACATTATCCTTAATACTGATGCTTATTGACTT ACTATGGGGTTACCTCTAGATAAATCCATAATAAGTTGAAAATATAAGTAAAAAATGC CCTTAATACACCTAACCTACCAAACATCATAGCTGAGCCCAGCCTGCCTTAGCTATGCT CAGACACTGACGTCAGCCTACAATTGGCAAAATCACACAGCAGCACAGTCTACTGCAG AGCATCTGCTGTTTGCCCTTGTGACTGCGTGGCTGCCTGGGAGCTTCCCAGCTTCACAA GACAGTATTACGTAGCACATCACTAGCCTGGGGAAAGATCAAAGTTGAAAATTTGAAG TGTGGTTTCCATTGAATGTGTACTGCTTTTGCACCATCATCAAGTCAAAAAATTTTAGTT GAACCAGCCTAAGTTTGGGACCATCTTTATTTTCAGGAGGAACTTCCATGTACATTGAT GACGGACGATAGAATCCGTTTCTATCATCCTAATGAACATAATGAATAAATCCAGACA AACATAAACATTAACAGAGTAAGCAGCTTTCGGGGCTGGAAGCCAGAAGAGGGTGGG AGCGCAGAGAGAGAGGCCAAACACCAGGGCTGCTTCTGCTTTGCGGGTATTTGCTGAT CTGGACA...

Claims

CLAIMS 1. A therapeutic nucleic acid capable of binding to a target indel allele associated with a pathogenic allele, wherein the pathogenic allele causes a dominant negative genetic disorder or gain-of function genetic disorder, and wherein the disorder is not Huntington’s disease.

2. The therapeutic nucleic acid of claim 1, wherein the target indel allele is on the same haplotype as the pathogenic allele.

3. The therapeutic nucleic acid of claim 1 or 2, wherein the target indel allele is present within pre-mRNA or mRNA encoding the pathogenic allele.

4. The therapeutic nucleic acid of any one of claims 1 to 3, wherein the target indel allele has a high minor allele frequency (MAF), preferably wherein the target indel allele has an MAF of at least 0.01, more preferably wherein the target indel allele has an MAF of about at least 0.06, at least 0.07, at least 0.08, at least 0.09, or at least 0.

1.

5. The therapeutic nucleic acid of any one of claims 1 to 4, wherein the target indel allele is an insertion and / or deletion of one or more base-pairs, preferably wherein the target indel allele is an insertion or deletion of between 1 and 50bp.

6. The therapeutic nucleic acid of any one of claims 1 to 5, wherein the target indel allele is present in a coding or non-coding region, preferably wherein the target indel allele is present in an intron and is an intronic indel allele.

7. The therapeutic nucleic acid of any one of claims 1 to 6, wherein the therapeutic nucleic acid comprises a target recognition sequence, wherein the target recognition sequence is at least partially complementary to a target nucleic acid sequence comprising the target indel allele.

8. The therapeutic nucleic acid of claim 7, wherein the target nucleic acid sequence is DNA, or RNA, preferably wherein the target nucleic acid sequence is pre-mRNA or mRNA.

9. The therapeutic nucleic acid of claim 7 or 8, wherein the target recognition sequence is between 15 to 30 nucleotides in length, preferably between 18 to 25 nucleotides in length.

10. The therapeutic nucleic acid of any one of claims 7 to 9, wherein the target recognition sequence has complementarity to the target nucleic acid sequence across at least 75%, at least80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% of its total length.

11. The therapeutic nucleic acid of any one of claims 1 to 10, wherein the therapeutic nucleic acid has a total length of between 10 to 120 nucleotides, preferably between 15 to 30 nucleotides.

12. The therapeutic nucleic acid of any one of claims 1 to 11, wherein the therapeutic nucleic acid is selected from ASO, siRNA, shRNA, gRNA and miRNA, preferably wherein the therapeutic nucleic acid is an antisense oligonucleotide (ASO).

13. The therapeutic nucleic acid of claim 12, wherein the ASO is a gapmer.

14. The therapeutic nucleic acid of any one of claims 1 to 13, wherein the pathogenic allele is a pathogenic allele which causes a cardiac disease, selected from: MYH7, ACTC1, ACTN2, AKAP9, ANKRD1, CACNA1C, CALM1, CALM2, CALM3, CAV3, CHRM2, COL1A2, CRYAB, CSRP3, DES, DSC2, EYA4, GATA4, GATAD1, JUP, KCNJ2, KCNJ5, LDB3, LRRC10, MURC / CAVIN4, MYL2, MYL3, MYLK2, MYOT, MYOZ2, MYPN, NEBL, NEXN, NKX2-5, PDLIM3, PLN, PRDM16, PRKAG2, PSEN1, PSEN2, RAF1, RBM20, RYR2, SCN4B, SNTA1, TBX20, TCAP, TMEM43, TNNC1, TNNI3, TNNT2, TOR1AIP1, TPM1, TRDN, TTR, TXNRD2, and VCL; or a pathogenic allele which causes a Skeletal Dysplasia or Bone Disorder selected from: ALK2 / ACVR1, CDC73, COL1A1, COL1A2, COL2A1, COMP, COMP, FGF23, FGF23, FGFR3, FGFR3, GCM2, MATN3, and MEN1; or a pathogenic allele which causes a Neurological and Neuromuscular Disorder selected from: AARS1, ADCY5, AFG3L2, ATL1, ATL3, ATN1, ATXN1, ATXN10, ATXN11, ATXN7, ATXN8, ATXN8OS, BEAN1, CACNA1A, CACNA1G, CCDC88C, CHCHD10, CHKB, CHMP2B, COL6A1, COL6A2, COL6A3, C9ORF72, DAB1, DCTN1, DLK1, DNM2, DNM2, ELOVL4, FAT2, FGF14, FTL, FUS, GABRG2, GARS1, GFAP, GNE, GRIN2B, GRN, GRM1, HCN1, HNRNPA1, HNRNPA1, HNRNPA2B1, HNRNPA2B1, HTT, HSPB1, HSPB8, HSPB8, ITPR1, JPH3, KCNA1, KCNC3, KCND3, KCNQ2, KCNQ2, KCNQ2, KCNQ3, KCNQ3, KISS1, KISS1R, KMT2B, MLC1, MKRN3, MFN2, MME, MORC2, MYHC2A, NKX2-1, NOTCH3, NOP56, PABPN1, PDE2A, PHIP, PLD3, PNKD, PPP2R2B, PRKCG, PRNP, PRNP, PRNP, PRRT2, PRRT2, RYR1, SAMD9L, SCN1A, SCN2A, SCN8A, SCN9A, SCN10A, SCN11A, SLC2A1, SOD1, SPTBN2, SPTLC1, SPTLC2, SQSTM1, STUB1, STX1B, TARDBP, TBP, TGM6, TRPC3, TRPV4, TTBK2, and VCP; or a pathogenic allele which causes a Haematological Disorder selected from: ALAS2, ELANE, EPAS1, GFI1, GP1BA, GP1BA, HAX1, HBA1, HBA2,HBB, MPL, PLAU, SLC25A38, SLFN14, and THPO; or a pathogenic allele which causes an Endocrine or Metabolic Disorder selected from: ABCB6, CACNA1H, CLCN2, CYP19A1, FGF23, FTL, KCNJ5, PLIN1, PPARG, PRSS1, SLC40A1, TSHR, TSHR, and TTR; or a pathogenic allele which causes an Eye Disorder selected from: BEST1, CAPN5, CFI, COL4A1, COL8A2, COL8A2, CTNNA1, EFEMP1, FSCN2, FZD4, GRHL2, IMPDH1, IMPG1, KLHL7, KRT12, MAPKAPK3, NR2E3, OPA3, OPN1LW, OPN1MW, OTX2, OVOL2, PAX6, PLA2G5, PRPF3, PRPF8, RGS9BP, RHO, RP1L1, RP9, SMCHD1, SNRNP200, TCF4, TGFBI, TOPORS, TREX1, VSX1, ZEB1, and ZNF408; or a pathogenic allele which causes a Renal Disorder selected from: ANGPT2, BSND, CLCNKB, FLT4, FXYD2, GJC2, KCNJ1, PIEZO1, and SLC12A1; or a pathogenic allele which causes an Immunological Disorder selected from: ADAMTS10, ADAMTS17, AP1S3, LTBP2, NLRP3, NOD2, PTPN22, TSC1, and TSC2; or other pathogenic allele selected from: APOE, ATP2A2, COL4A1, GNAS, INHBE, KRT14, KRT5, NKX2-1, SLC22A1, PAFAH1B1, PMVK, PRNP, PSTPIP1, STX16, and TTR; preferably wherein the pathogenic allele is selected from a pathogenic: TNNI3, MYH7, and TNNT2 allele.

15. The therapeutic nucleic acid of claim 14, wherein the pathogenic allele is a pathogenic MYH7 allele and the target indel allele is an allele of intronic indel: rs34598192.

16. The therapeutic nucleic acid of claim 15, wherein the therapeutic nucleic acid comprises a sequence selected from SEQ ID NOs: 33-84, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, preferably wherein the therapeutic nucleic acid comprises a sequence selected from SEQ ID NOs: 33-78, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, preferably wherein the therapeutic nucleic acid comprises a sequence according to SEQ ID 66 (D7) , SEQ ID NO: 33 (C1), or SEQ ID 36 (C4), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto.

17. The therapeutic nucleic acid of claim 14, wherein the pathogenic allele is a pathogenic TNNI3 allele and the target indel allele is an allele of intronic indel rs377373012.

18. The therapeutic nucleic acid of claim 17, wherein the therapeutic nucleic acid comprises a sequence selected from SEQ ID NOs: 85-145, 260, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, preferably wherein the therapeutic nucleic acid comprises a sequence selected from SEQ ID NOs: 85-139, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, preferably wherein the therapeutic nucleic acid comprises a sequence according to SEQ ID NO: 87 (E3) or SEQ ID NO: 101 (E17), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto.

19. The therapeutic nucleic acid of claim 14, wherein the pathogenic allele is a pathogenic TNNT2 allele and the target indel allele is an allele of intronic indel rs45533739.

20. The therapeutic nucleic acid of claim 19, wherein the therapeutic nucleic acid comprises a sequence selected from SEQ ID NOs: 146-187, 232, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, preferably wherein the therapeutic nucleic acid comprises a sequence selected from SEQ ID NOs: 146-181, 232, or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto, preferably wherein the therapeutic nucleic acid comprises a sequence according to SEQ ID NO: 156 (F12), or a sequence having at least 70% identity thereto, optionally at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or up to 100% identity thereto.

21. The therapeutic nucleic acid of any one of claims 1 to 20, wherein the dominant negative genetic disorder or gain-of function genetic disorder is a cardiomyopathy, preferably selected from: hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM), and restrictive cardiomyopathy (RCM).

22. A conjugate comprising the therapeutic nucleic acid of any of claims 1 to 21 covalently linked to a delivery group.

23. The conjugate of claim 22 wherein the delivery group is selected from: an antibody, a peptide, small molecule chemical, polymer, and lipid, preferably wherein the delivery group is an antibody.

24. The conjugate of claim 22 or 23 wherein the delivery group is covalently linked to the therapeutic nucleic acid via a linker.

25. A delivery particle comprising the therapeutic nucleic acid of any of claims 1-21, or the conjugate of any of claims 22-24.

26. The delivery particle of claim 25, wherein the delivery particle is selected from: a nanoparticle, a liposome, and an exosome.

27. A pharmaceutical composition comprising the therapeutic nucleic acid according to any of claims 1-21, the conjugate according to any of claims 22-24, or the delivery particle of claims 25 or 26, and one or more pharmaceutically acceptable excipients.

28. A therapeutic nucleic acid according to any of claims 1-21, conjugate according to any of claims 22-24, delivery particle according to claim 25 or 26, or pharmaceutical composition according to claim 27, for use in the treatment or prevention of a dominant negative or gain-of- function genetic disorder in a subject, wherein the disorder is not Huntington’s disease.

29. The therapeutic nucleic acid, conjugate, delivery particle or pharmaceutical composition for use according to claim 28, wherein the dominant negative or gain-of-function genetic disorder is a cardiomyopathy, preferably selected from: hypertrophic cardiomyopathy (HCM), dilated cardiomyopathy (DCM) and restrictive cardiomyopathy (RCM).

30. The therapeutic nucleic acid, conjugate, delivery particle or pharmaceutical composition for use according to claim 28 or 29, wherein the treatment or prevention is by allele-specific knockdown of a pathogenic allele which causes the dominant negative or gain-of-function genetic disorder.

31. The therapeutic nucleic acid, conjugate, delivery particle or pharmaceutical composition for use according to any one of claims 28 to 30, wherein the treatment reduces the level, expression and / or activity of transcripts of a pathogenic allele which causes the dominant negative or gain- of-function genetic disorder.

32. A therapeutic nucleic acid which is capable of binding to a target indel allele of an intronic indel selected from: rs59464879, rs751205475 and rs78373442 associated with a pathogenic HTT allele which causes Huntington’s disease.

33. A therapeutic nucleic acid according to claim 32 for use in the treatment or prevention of Huntington’s disease.

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