Compositions for and methods of engineering the transcriptome

Abstract

Disclosed herein are CRISPR-free compositions for and methods of generating chimeric RNA molecules via trans-splicing and methods of treating and / or preventing a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations) using chimeric RNA molecules generated via trans-splicing.

Description

COMPOSITIONS FOR AND METHODS OF ENGINEERING THE TRANSCRIPTOME I. CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 763,378 filed 26 February72025, which is incorporated herein in its entirety.II. REFERENCE TO THE SEQUENCE LISTING

[0002] The Sequence Listing submitted 26 February 2026 as an XML file named “25-4049-WO_SL”, created on 26 February 2026 and having a size of 180,224 bytes is hereby incorporated by reference pursuant to 37 C.F.R. § 1.52(e)(5).HI. BACKGROUND

[0003] In mammalian cells, mutations or aberrations in transcriptionally active regions of chromosomal DNA give rise to pre-mRNA bearing identical mutations or aberrations. If the mutation is located in a non-coding region, then processing of the pre-mRNA may be altered or abolished. If the mutation is located in an exonic region of the pre-mRNA, then that mutation will be passed to the mature mRNA sequence. These mutations or aberrations can contribute to inhibition of complete protein translation of the encoded protein (non-sense mutation) or modify the primary structure of the encoded protein in a counter-productive manner (missense mutation). Collectively, these genetically encoded mutations or aberrations may function to contribute to pathogenesis in eukaryotes.

[0004] The field of gene therapy has aimed to correct such genetic abnormalities through adoptive gene transfer of recombinant nucleic acids bearing a sequence capable of producing the protein product of the mutated gene. This strategy, conventionally termed “classical gene therapy” has proven to be a safe and effective strategy' for phenotypic correction of genetic disorders, with several gene therapy products available on the market. However, current approaches to gene therapy are limited due to toxicity arising from over-expression of transcripts, the inability to deliver large transcripts exceeding the packaging capacity' of AAV vectors (i.e., considered to be the standard vector for gene therapy and having a packaging capacity of ~4.7 KB), the inability to correct and replace large stretches of disease causing mRNA, and the inability to correct and replace autosomal dominant inheritance with a single transcript and / or approach.

[0005] Thus, there remains an urgent need for a minimally invasive, definitive therapy to address the underlying cause of as well as the sequelae of symptoms associated with these various genetic diseases and disorders. Consequently, the present disclosure provides compositions for and methods of generating chimeric RNA molecules via trans-splicing (and without CRISPR) and treating and / or preventing a genetic disease and / or disorder, which can be used alone or in combination with other treatments.IV. BRIEF DESCRIPTION OF THE FIGURES

[0006] FIG. 1A shows a schematic of the 3’ trans-splicing approach while FIG. IB shows a schematic of the 5 ’ trans-splicing approach.

[0007] FIG. 2 shows a schematic of the split green fluorescent protein reporter and the guide position along the MYH7 intron. Guide 1 targets the final 50 bases of MYH7 intron 22 / 23 and progress in reverse along the intron in 20 nucleotide increments until Guide 29 (SEQ ID NO:01 -SEQ ID NO:29).

[0008] FIG. 3 shows EGFP rescue as measured by flow cytometry for each guide. EGFP rescue is a proxy for trans-splicing efficiency. Plasmids with a CMV promoter driving expression of the trans-splicing RNA were transfected into HEK293 cells with splitGFP reporters (FIG. 2) containing the human MYH7 intron 22 / 23. For 5’ trans-splicing, each trans-splicing RNA contained the 5’ open reading frame of EGFP (SEQ ID NO:93) that was operably linked minimal hemi intron (SEQ ID NO:91) that was operably linked to an antisense binding region (SEQ ID NO:01 - SEQ ID NO:29). For 3’ trans-splicing, each trans-splicing RNA contained an antisense binding region (SEQ ID NO:01 - SEQ ID NO:29) that was operably linked minimal hemi intron (SEQ ID NO:90) that was operably linked the 3’ open reading frame of EGFP (SEQ ID NO:94).

[0009] FIG. 4 shows trans-splicing efficiency as a function of guide length. EGFP rescue was measured by flow cytometry for each guide. EGFP rescue is a proxy for trans-splicing efficiency. Plasmids with a CMV promoter driving expression of the trans-splicing RNA were transfected into HEK293 cells with splitGFP reporters (FIG. 2) containing the human MYH7 intron 22 / 23. For 5’ trans-splicing, each trans-splicing RNA contained the 5’ open reading frame of EGFP (SEQ ID NO:93) that was operably linked minimal hemi intron (SEQ ID NO:91) that was operably linked to an antisense binding region (SEQ ID NO:37 - SEQ ID NO:43). For 3’ trans-splicing, each trans-splicing RNA contained an antisense binding region (SEQ ID NO:30 - SEQ ID NO:36) that was operably linked minimal hemi intron (SEQ ID NO:90) that was operably linked the 3’ open reading frame of EGFP (SEQ ID NO:94).

[0010] FIG. 5 shows the effect of guide proximal RNA structures on trans-splicing efficiency. EGFP rescue was measured by flow cytometry for each guide. EGFP rescue is a proxy for trans-splicing efficiency. Plasmids with a CMV promoter driving expression of the trans-splicing RNA were transfected into HEK293 cells with splitGFP reporters (FIG.2) containing the human MYH7 intron 22 / 23. For 5' trans-splicing, each trans-splicing RNA contained the 5’ open reading frame of EGFP (SEQ ID NO:93) that was operably linked minimal hemi intron (SEQ ID NO:91) that was operably linked to an antisense binding region (SEQ ID NO:38, SEQ ID NO:46, or SEQ ID NO:47). For 3’ trans-splicing, each trans-splicing RNA contained an antisense binding region (SEQ ID NO:31, SEQ ID NO:44, or SEQ ID NO:45) that was operably linked minimal hemiintron (SEQ ID NO: 90) that was operably linked the 3’ open reading frame of EGFP (SEQ ID NO:94).

[0011] FIG.6 shows schematic of the multivalent guide approach to 5’ and 3’ trans-splicing. For 5’ trans-splicing, Guide 5 and Guide 19 were combined to make multivalent guide trans-splicing editor with multivalent guides. For 3’ trans-splicing. Guide 16 and Guide 26 were combined to make multivalent guide trans-splicing editor with multivalent guides (SEQ ID NO:48 and SEQ ID NO:58).

[0012] FIG. 7 shows effect of multivalent guide on trans-splicing efficiency. EGFP rescue was measured by flow cytometry for each guide. EGFP rescue is a proxy for trans-splicing efficiency. Plasmids with a CMV promoter driving expression of the trans-splicing RNA were transfected into HEK293 cells with splitGFP reporters (FIG. 2) containing the human MYH7 intron 22 / 23. For 5’ trans-splicing, each trans-splicing RNA contained the 5’ open reading frame of EGFP (SEQ ID NO:93) that was operably linked minimal hemi intron (SEQ ID NO:91) that was operably linked to an antisense binding region (SEQ ID NO: 38 or SEQ ID NO: 50). For 3’ trans-splicing, each trans-splicing RNA contained an antisense binding region (SEQ ID NO:31 or SEQ ID NO:48) that was operably linked minimal hemi intron (SEQ ID NO:90) that was operably linked the 3’ open reading frame of EGFP (SEQ ID NO: 94).

[0013] FIG. 8A - FIG. 8B shows EGFP rescue via trans-splicing at escalating MOI of self-complementary AAV2 packaging the 3’ multivalent guide. FIG. 8A and FIG. 8B show %GFP Positive cells as a function of MOI. EGFP rescue was measured by flow cytometry for each guide. EGFP rescue is a proxy for trans-splicing efficiency. AAV packaging a genome with a CMV promoter driving expression of the trans-splicing RNA were transduced into HEK293 cells that stably express a genome integrated splitGFP reporter (FIG.2) containing the human MYH7 intron 22 / 23. Each 3’ trans-splicing RNA contained an antisense binding region (SEQ ID NO:48) that was operably linked minimal hemi intron (SEQ ID NO:90) that was operably linked the 3’ open reading frame of EGFP (SEQ ID NO:94).

[0014] FIG. 9 shows a schematic of the optimized MYH7 targeting 3’ trans-splicing RNA splicing into the split green fluorescent reporter. Detailed on the schematic are primers across the splice junction to amplify the trans-spliced product (fusion of SEQ ID NO:93 followed by SEQ ID NO:96). Here, RAFT is RNA Assisted Fragment Trans-splicing.

[0015] FIG. 10 shows a DNA gel of PCR products amplifying the splice junction of the chimeric RNA formed between the split green fluorescent reporter and the MYH7 targeting 3’ trans splicing RNA. Amplicons were generated from RNA harvested from HEK293 cells that stably express the reporter and were transduced with 0 (lane 2), le4 vg / cell (lane 3), le5 vg / cell (lane 4), and 5e5 vg / cell (lane 5) of AAV2 expressing the trans-splicing RNA.

[0016] FIG. 11 shows a schematic of the optimized MYH7 targeting 3’ trans-splicing RNA splicing into the endogenous MYH7 pre-mRNA.

[0017] FIG. 12 shows a DNA gel of PCR products amplifying the splice junction of the chimeric RNA formed between the MYH7 mRNA and the MYH7 targeting 3’ trans-splicing RNA containing the second half of the green fluorescent protein open reading frame (Lanes 5-7). Amplicons were generated from RNA harvested from HEK293 cells that stably expressed MYH7 via CRISPR activation.

[0018] FIG. 13 shows Sanger sequencing of -300 bp amplicons extracted from the gel in FIG.14 confirm precise trans-splicing (SEQ ID NQ:107, SEQ ID NO: 108, SEQ ID NO: 109). Alignment is to exon 22 of MYH7 followed by the open reading frame of the second half of enhanced green fluorescent protein.

[0019] FIG. 14 shows schematic of the “blocking” U7 snRNAs in combination with 5’ and 3’ trans-splicing. U7 snRNAs positions are labeled 1-10 above the split green fluorescent reporter.

[0020] FIG. 15 shows the effect of “blocking” U7 snRNAs (SEQ ID NO:62 - SEQ ID NO:71) on trans-splicing efficiency. EGFP rescue was measured by flow cytometry for each guide. EGFP rescue is a proxy for trans-splicing efficiency. Guide numbers correspond to the numbered positions of each U7 snRNA in FIG. 14, and Aflll denotes a U7 snRNA that has an Aflll restriction site in the place of its anti-sense domain. Plasmids containing a “blocking” U7 snRNA transgene (SEQ ID NO:72 - SEQ ID NO:82) were co-transfected into HEK293 cells with a trans-splicing RNA and splitGFP reporters (FIG.2) containing the human MYH7 intron 22 / 23. For 5’ trans-splicing, each trans-splicing RNA contained the 5’ open reading frame of EGFP (SEQ ID NO:93) that was operably linked minimal hemi intron (SEQ ID NO:91) that was operably linked to an antisense binding region (SEQ ID NO:50). For 3’ trans-splicing, each trans-splicing RNA contained an antisense binding region (SEQ ID NO:48) that was operably linked minimal hemi intron (SEQ ID NO:90) that was operably linked the 3’ open reading frame of EGFP (SEQ ID NO:94).

[0021] FIG. 16A - FIG. 16B show the effect of multivalent guide on trans-splicing efficiency. EGFP rescue was measured by flow cytometry for each guide. EGFP rescue is a proxy for trans-splicing efficiency. Plasmids with a CMV promoter driving expression of the trans-splicing RNA were transfected into HEK293 cells with splitGFP reporters (FIG.2) containing the human MYH7 intron 22 / 23. Each 3’ trans-splicing RNA contained an antisense binding region that was operably linked minimal hemi intron (SEQ ID NQ:90) that was operably linked the 3’ open reading frame of EGFP (SEQ ID NO:94). Single 50 bp length anti-sense binding region guides 16 (SEQ ID NO: 16) or 25 (SEQ ID NO:25) were compared to the optimized multivalent guide architecture(SEQ ID NO:48) and alternative multivalent guide architectures that change the order of SLBP and TAR hairpins or the order of the anti-sense binding regions.

[0022] FIG. 17A- FIG. 17B show an assay of multivalent guide architectures on 5’ trans-splicing efficiency as measured by %GFP Positive (FIG. 17A) and by MFI (FIG. 17B). EGFP rescue was measured by flow cytometry for trans-splicing RNA. EGFP rescue is a proxy for trans-splicing efficiency. Plasmids with a CMV promoter driving expression of the trans-splicing RNA were transfected into HEK293 cells with splitGFP reporters (FIG. 2) containing the human MYH7 intron 22 / 23. Briefly, each trans-splicing RNA contained the 5’ open reading frame of EGFP (SEQ ID NO: 93) that was operably linked minimal hemi intron (SEQ ID NO:91) or ahemi intron with additional U1 binding sites (SEQ ID NO:92) that was operably linked to one of several multivalent guide architectures (SEQ ID NO:49 - SEQ ID NO:60).

[0023] FIG. 18A - FIG. 18B show the effect of multivalent guide on trans-splicing efficiency as measured by %GFP Positive (FIG. 18A) and by MFI (FIG. 18B). EGFP rescue was measured by flow cytometry for each guide. EGFP rescue is a proxy for trans-splicing efficiency. Plasmids with a CMV promoter driving expression of the trans-splicing RNA were transfected into HEK293 cells that stably express a genome integrated splitGFP reporter (FIG. 2) containing the human MYH7 intron 22 / 23. Each 3’ trans-splicing RNA contained an antisense binding region (SEQ ID NO:30, SEQ ID NO:31, or SEQ ID NO:48) that was operably linked minimal hemi intron (SEQ ID NO:90) that was operably linked the 3’ open reading frame of EGFP (SEQ ID NO:94).

[0024] FIG. 19 shows RNA editing was quantified in HEK293 cells that were engineered with CRISPR activation to express the endogenous MYH7 RNA. Plasmid (SEQ ID NO: 85) encoding the 3 ’ trans-splicing RNA was transfected into these cells and cells were transduced with AAV2 containing a genome expressing the optimal 3’ trans-splicing RNA (SEQ ID NO:84). 72 hours later RNA was harvested and converted to cDNA followed by targeted amplicon sequencing of the exon 22 / 23 splice junction to quantify trans-splicing efficiency.V. BRIEF SUMMARY

[0025] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be transspliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.

[0026] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5‘ hemi intron linked to an exogenousRNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA.

[0027] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA.

[0028] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.

[0029] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5' portion of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA.

[0030] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ‘ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA.

[0031] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0032] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0033] Disclosed herein is a nucleic acid molecule an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be transspliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0034] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0035] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ' portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0036] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0037] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA.

[0038] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA.

[0039] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA.

[0040] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA.

[0041] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; a guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA.

[0042] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA.

[0043] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA.

[0044] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous premRNA. Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0045] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0046] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0047] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0048] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0049] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenousMYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0050] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0051] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3 ’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3' portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0052] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0053] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0054] Disclosed herein is a method of generating a chimeric RNA molecule in a cell, the method comprising contacting an endogenous pre-mRNA in a cell with a disclosed 5’ replacement construct, wherein the resulting chimeric RNA transcript comprises the 3’ portion of the targeted endogenous pre-mRNA and the 5' portion of the exogenous RNA.

[0055] Disclosed herein is a method of generating a chimeric RNA molecule, the method comprising contacting one or more cells with a disclosed 5’ replacement construct, wherein the resulting chimeric RNA transcript comprises the 3’ portion of a targeted endogenous pre-mRNA and the 5’ portion of the exogenous RNA. Disclosed herein is a method of generating a chimeric RNA molecule, the method comprising contacting one or more cells with a disclosed nucleic acid molecule comprising a disclosed 5’ replacement construct, wherein the resulting chimeric RNA transcript comprises the 3’ portion of the targeted endogenous pre-mRNA and the 5’ portion of the exogenous RNA.

[0056] Disclosed herein is a method of generating a chimeric RNA molecule, the method comprising contacting one or more cells with a disclosed viral vector or non-viral vector comprising a disclosed 5’ replacement construct, wherein the resulting chimeric RNA transcript comprises the 3’ portion of the targeted endogenous pre-mRNA and the 5’ portion of the exogenous RNA.

[0057] Disclosed herein is a method of generating a chimeric RNA molecule, the method comprising contacting one or more cells with a disclosed pharmaceutical formulation comprising a disclosed 5’ replacement construct, wherein the resulting chimeric RNA transcript comprises the 3’ portion of the targeted endogenous pre-mRNA and the 5‘ portion of the exogenous RNA.

[0058] Disclosed herein is a method of generating a chimeric RNA molecule in a cell, the method comprising contacting an endogenous pre-mRNA in a cell with a disclosed 3’ replacement construct, wherein the resulting chimeric RNA transcript comprises the 5’ portion of the targeted endogenous pre-mRNA and the 3’ portion of the exogenous RNA.

[0059] Disclosed herein is a method of generating a chimeric RNA molecule, the method comprising contacting one or more cells with a disclosed 3’ replacement construct, wherein the resulting chimeric RNA transcript can comprise the 5’ portion of the targeted endogenous pre-mRNA and the 3’ portion of the exogenous RNA.

[0060] Disclosed herein is a method of generating a chimeric RNA molecule, the method comprising contacting one or more cells with a disclosed nucleic acid molecule comprising a disclosed 3’ replacement construct wherein the resulting chimeric RNA transcnpt comprises the 5’ portion of the targeted endogenous pre-mRNA and the 3’ portion of the exogenous RNA.

[0061] Disclosed herein is a method of generating a chimeric RNA molecule, the method comprising contacting one or more cells with a viral vector or a non-viral vector comprising adisclosed 3’ replacement construct wherein the resulting chimeric RNA transcript comprises the 5’ portion of the targeted endogenous pre-mRNA and the 3’ portion of the exogenous RNA.

[0062] Disclosed herein is a method of generating a chimeric RNA molecule, the method comprising contacting one or more cells with a disclosed pharmaceutical formulation comprising a disclosed 3 ’ replacement construct wherein the resulting chimeric RNA transcript comprises the 5’ portion of the targeted endogenous pre-mRNA and the 3’ portion of the exogenous RNA.

[0063] Disclosed herein is a method of treating a subject, the method comprising generating a chimeric RNA molecule in one or more cells by administering to a subject having hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations a therapeutically effective amount of a disclosed non-viral vector or a disclosed viral vector, wherein the resulting chimeric RNA molecule can restore one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation, wherein the subject is treated by the slowing of disease progression.

[0064] Disclosed herein is a method of treating a subject the method comprising generating a chimeric RNA molecule in one or more cells by administering to a subject having hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations a therapeutically effective amount of a disclosed non-viral vector or a disclosed viral vector, wherein the resulting chimeric RNA molecule can restore the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme, wherein the subject is treated by the slowing of disease progression.

[0065] Disclosed herein is a method of treating a subject, the method comprising generating a chimeric RNA molecule in one or more cells by administering to a subject having hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations a therapeutically effective amount of a disclosed pharmaceutic formulation, wherein the resulting chimeric RNA molecule can restore one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation, wherein the subject is treated by the slowing of disease progression.

[0066] Disclosed herein is a method of treating a subject the method comprising generating a chimeric RNA molecule in one or more cells by administering to a subject having hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations a therapeutically effective amount of a disclosed pharmaceutical formulation, wherein the resulting chimeric RNA molecule can restore the functionality’ and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme, wherein the subject is treated by the slowing of disease progression.VI. DETAILED DESCRIPTION

[0067] The present disclosure describes formulations, compounded compositions, kits, capsules, containers, and / or methods thereof. It is to be understood that the inventive aspects of which arenot limited to specific synthetic methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, example methods and materials are now described.

[0068] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention.A. Definitions

[0069] Before the present compounds, compositions, articles, systems, devices, and / or methods are disclosed and descnbed, it is to be understood that they are not limited to specific synthetic methods unless otherwise specified, or to particular reagents unless otherwise specified, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, example methods and materials are now described.

[0070] This disclosure describes inventive concepts with reference to specific examples. However, the intent is to cover all modifications, equivalents, and alternatives of the inventive concepts that are consistent with this disclosure.

[0071] As used in the specification and the appended claims, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise.

[0072] The phrase “consisting essentially of’ limits the scope of a claim to the recited components in a composition or the recited steps in a method as well as those that do not materially affect the basic and novel characteristic or characteristics of the claimed composition or claimed method. The phrase “consisting of’ excludes any component, step, or element that is not recited in the claim. The phrase “comprising” is synonymous with “including”, “containing”, or “characterized by”, and is inclusive or open-ended. “Comprising” does not exclude additional, unrecited components or steps.

[0073] In an aspect, when referring to any numerical value, the term “about” means a value falling within a range that is ± 10% of the stated value.

[0074] Ranges can be expressed herein as from “about” one particular value, and / or to “about” another particular value. When such a range is expressed, a further aspect includes from the oneparticular value and / or to the other particular value. Similarly, when values are expressed as approximations, by use of the antecedent '‘about,” it will be understood that the particular value forms a further aspect. It will be further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint and independently of the other endpoint. It is also understood that there are a number of values disclosed herein, and that each value is also herein disclosed as ‘'about” that particular value in addition to the value itself. For example, if the value “10” is disclosed, then “about 10” is also disclosed. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0075] References in the specification and concluding claims to parts by weight of a particular element or component in a composition denotes the weight relationship between the element or component and any other elements or components in the composition or article for which a part by weight is expressed. Thus, in a compound containing 2 parts by weight component X and 5 parts by weight component Y, X and Y are present at a weight ratio of 2:5, and are present in such ratio regardless of whether additional components are contained in the compound.

[0076] In an aspect, the terms “optional” or “optionally” means that the subsequently described event or circumstance can or cannot occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. In an aspect, a disclosed method can optionally comprise one or more additional steps, such as, for example, repeating an administering step or altering an administering step.

[0077] In an aspect, “isolated” refers to a nucleic acid molecule or a nucleic acid sequence that has been substantially separated, produced apart from, or purified away from other biological components in the cell or tissue of an organism in which the component occurs, such as other cells, chromosomal and extrachromosomal DNA and RNA, and proteins. Nucleic acids and proteins that have been “isolated” include nucleic acids and proteins purified by standard purification methods. The term also embraces nucleic acids and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acids and proteins.

[0078] In an aspect, the term “diagnosed” means having been subjected to an examination by a person of skill, for example, a physician, and found to have a condition that can be diagnosed or treated by one or more of the disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof, or by one or more of the disclosed methods. For example, “diagnosed with a disease or disorder” means having been subjected to an examination by a person of skill, for example, a physician, and found to have a condition (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations) that can be treated by one or more of the disclosed nucleic acid molecules, disclosed vectors, disclosedpharmaceutical formulations, or a combination thereof, or by one or more of the disclosed methods. For example, ‘'suspected of having a disease or disorder” can mean having been subjected to an examination by a person of skill, for example, a physician, and found to have a condition (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations) that can likely be treated by one or more of by one or more of the disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof, or by one or more of the disclosed methods. In an aspect, an examination can be physical, can involve various tests (e.g., blood tests, genotyping, biopsies, etc.) and assays (e.g., enzy matic assay), or a combination thereof.

[0079] In an aspect, the term “prevent” or “preventing” or “prevention” refers to precluding, averting, obviating, forestalling, stopping, or hindering something from happening, especially by advance action. It is understood that yvhere reduce, inhibit, or prevent are used herein, unless specifically indicated otherwise, the use of the other two words is also expressly disclosed. In an aspect, preventing a disease or disorder having chromatin deregulation and / or chromatin dysregulation is intended. The words '‘prevent”, “preventing”, and “prevention” also refer to prophylactic or preventative measures for protecting or precluding a subject (e.g., an individual) not having a given a disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations) or related complication from progressing to that complication.

[0080] In an aspect, the skilled person can determine an efficacious dose, an efficacious schedule, and an efficacious route of administration for one or more of the disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof to treat or prevent a disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations). In an aspect, the skilled person can also alter, change, or modify an aspect of an administering step to improve efficacy of one or more of the disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof.

[0081] By “determining the amount” is meant both an absolute quantification of a particular analyte (e g., an mRNA sequence containing a particular tag) or a determination of the relative abundance of a particular analyte (e.g., an amount as compared to a mRNA sequence including a different tag). The phrase includes both direct or indirect measurements of abundance (e.g., individual mRNA transcripts may be quantified or the amount of amplification of an mRNA sequence under certain conditions for a certain period may be used a surrogate for individual transcript quantification) or both.

[0082] In an aspect, the term “pharmaceutically acceptable carrier” refers to sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, as well as sterile powders forreconstitution into sterile injectable solutions or dispersions just prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents, or vehicles include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), carboxymethylcellulose and suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. In an aspect, a pharmaceutical carrier employed can be a solid, liquid, or gas. In an aspect, examples of solid carriers can include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. In an aspect, examples of liquid carriers can include sugar syrup, peanut oil, olive oil, and water. In an aspect, examples of gaseous carriers can include carbon dioxide and nitrogen. In preparing a disclosed composition for oral dosage form, any convenient pharmaceutical media can be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like can be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like can be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets can be coated by standard aqueous or nonaqueous techniques. Proper fluidity can be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants. These compositions can also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms can be ensured by the inclusion of various antibacterial and antifungal agents such as paraben, chlorobutanol, phenol, sorbic acid and the like. It can also be desirable to include isotonic agents such as sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the inclusion of agents, such as aluminum monostearate and gelatin, which delay absorption. Injectable depot forms are made by forming microencapsule matrices of the drug in biodegradable polymers such as polylactide-polyglycolide, poly(orthoesters) and poly(anhydrides). Depending upon the ratio of drug to polymer and the nature of the particular polymer employed, the rate of drug release can be controlled. Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues. The injectable formulations can be sterilized, for example, by filtration through a bacterial -retaining filter or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable media just prior to use. Suitable inert carriers can include sugars such as lactose. Desirably, at least 95% by weight of the particles of the active ingredient have an effective particle size in the range of 0.01 to 10 micrometers.

[0083] In an aspect, the term "excipient" refers to an inert substance which is commonly used as a diluent, vehicle, preservative, binder, or stabilizing agent, and includes, but is not limited to, proteins (e.g., serum albumin, etc.), amino acids (e.g., aspartic acid, glutamic acid, lysine, arginine, glycine, histidine, etc.), fatty acids and phospholipids (e.g., alkyl sulfonates, caprylate, etc.), surfactants (e.g.. SDS, polysorbate, nonionic surfactant, etc.), saccharides (e.g., sucrose, maltose, trehalose, etc.) and polyols (e.g., mannitol, sorbitol, etc.). See, also, for reference. Remington’s Pharmaceutical Sciences, (1990) Mack Publishing Co., Easton, Pa., which is hereby incorporated by reference in its entirety.

[0084] In an aspect, “concurrently” means (1) simultaneously in time, or (2) at different times during the course of a common treatment schedule.

[0085] In an aspect, small nuclear ribonucleoproteins (snRNPs) are complexes composed of small nuclear RNA (snRNA) and proteins in specific structures. Some of these snRNAs were uridine rich compared to ribosomal or messenger RNAs and thus were described as U snRNAs. These U snRNAs were further numbered (e.g., U1 snRNA) by their order of discovery, and not by size, location or abundance. The spliceosomes, comprising large complexes that catalyze splicing, are divided into major and minor spliceosomes: Ul, U2, U4, U5, U6 and Uli, U12, U4atac, U5atac and U6atac snRNP, respectively. Exceptionally, one of the U snRNPs, U7 snRNP, is not involved in splicing but is a key factor in the unique 3' end processing of replication-dependent histone (RDH) pre-mRNAs. Moreover, U7 snRNA is an important tool in therapeutic studies, with reports based on modified U7 snRNP (U7 Sm OPT) targeting splicing to induce efficient skipping or inclusion of selected exons. U7 Sm OPT is designed by changing the histone binding sequence at the 5' region of U7 snRNA to the complementary sequence of the gene to be modified. Further modifications include changing of U7 snRNP specific proteins. LsmlO and Lsmll, to the consensus protein ring of spliceosomal snRNPs. In U7 Sm OPT-based therapy, an antisense oligonucleotide is incorporated into the U7 snRNA. For this purpose, as a tool of manipulation of pre-mRNA splicing, antisense oligonucleotides are used in the U7 Sm OPT therapeutic strategy.

[0086] In an aspect, the term “contacting” can refer to bringing one or more of the disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof together with a target area or intended target area in such a manner that the one or more of the disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations, or a combination thereof exert an effect on the intended target or targeted area either directly or indirectly. A target area can comprise one or more cells, and in an aspect, one or more cells can be in a subject. A target area or intended target area can be one or more of a subject’s organs (e.g., lungs, heart, liver, kidney, brain, etc.). In an aspect, a target area or intended targetarea can be any cell or any organ infected by a disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations). In an aspect, a target area or intended target area can be any organ, tissue, or cells that are affected by a disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations).

[0087] In an aspect, “effective amount” and “amount effective” can refer to an amount that is sufficient to achieve the desired result such as, for example, the treatment and / or prevention of a disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations) or a suspected disease or disorder. In an aspect, the terms “effective amount” and “amount effective” can refer to an amount that is sufficient to achieve the desired an effect on an undesired condition (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations). For example, a “therapeutically effective amount” refers to an amount that is sufficient to achieve the desired therapeutic result or to have an effect on undesired symptoms, but is generally insufficient to cause adverse side effects. In an aspect, “therapeutically effective amount” means an amount of a disclosed nucleic acid molecule, a disclosed vector, or a disclosed pharmaceutical formulation; that (i) treats the particular disease, condition, or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations), (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations), or (iii) delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations). The specific therapeutically effective dose level for any particular patient will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the disclosed nucleic acid molecules, disclosed vectors, disclosed pharmaceutical formulations employed; the disclosed methods employed; the age, body weight, general health, sex and diet of the patient; the time of administration; the route of administration; the rate of excretion of the disclosed nucleic acid molecules, disclosed vectors, or disclosed pharmaceutical formulations employed; the duration of the treatment; drugs used in combination or coincidental with the disclosed nucleic acid molecules, disclosed vectors, or disclosed pharmaceutical formulations employed, and other like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the disclosed nucleic acid molecules, disclosed vectors, or disclosed pharmaceutical formulations at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. If desired, then the effective daily dose can be divided into multiple doses for purposes of administration. Consequently, a single dose of the disclosed nucleic acid molecules,disclosed vectors, or disclosed pharmaceutical formulations can contain such amounts or submultiples thereof to make up the daily dose. The dosage can be adjusted by the individual physician in the event of any contraindications. Dosage can var7, and can be administered in one or more dose administrations daily, for one or several days. Guidance can be found in the literature for appropriate dosages for given classes of pharmaceutical products. In further various aspects, a preparation can be administered in a '‘prophylactically effective amount”; that is, an amount effective for prevention of a disease or condition, such as, for example, a disease or disorder due to a missing, deficient, and / or mutant protein or enzyme (i.e. MYH7).

[0088] In an aspect, “RNA therapeutics” can refer to the use of oligonucleotides to target RNA. RNA therapeutics can offer the promise of uniquely targeting the precise nucleic acids involved in a particular disease with greater specificity, improved potency, and decreased toxicity. This could be particularly powerful for genetic diseases where it is most advantageous to aim for the RNA as opposed to the protein. In an aspect, a therapeutic RNA can comprise one or more expression sequences. As known to the art, expression sequences can comprise an RNAi, shRNA, mRNA, non-coding RNA (ncRNA), an antisense such as an antisense RNA, miRNA, morpholino oligonucleotide, peptide-nucleic acid (PNA) or ssDNA (with natural, and modified nucleotides, including but not limited to, LNA, BNA, 2’-0-Me-RNA, 2’-MEO-RNA, 2'-F-RNA), or analog or conjugate thereof. In an aspect, a disclosed therapeutic RNA can comprise one or more long non-coding RNA (IncRNA), such as, for example, a long intergenic non-coding RNA (lincRNA), pre-transcript, pre-miRNA, pre-mRNA, competing endogenous RNA (ceRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), pseudo-gene, rRNA, or tRNA. In an aspect, ncRNA can be piwi-interacting RNA (piRNA), primary miRNA (pri-miRNA), or premature miRNA (pre-miRNA). In an aspect, a disclosed therapeutic RNA or an RNA therapeutic can comprise antisense oligonucleotides (ASOs) that inhibit mRNA translation, oligonucleotides that function via RNA interference (RNAi) pathway, RNA molecules that behave like enzymes (ribozymes), RNA oligonucleotides that bind to proteins and other cellular molecules, and ASOs that bind to mRNA and form a structure that is recognized by RNase H resulting in cleavage of the mRNA target. In an aspect, RNA therapeutics can comprise RNAi and ASOs that inhibit mRNA translation. Generally speaking, as known to the art, RNAi operates sequence specifically and post-transcriptionally by activating ribonucleases which, along with other enzymes and complexes, coordinately degrade the RNA after the original RNA target has been cut into smaller pieces while antisense oligonucleotides bind to their target nucleic acid via Watson-Crick base pairing, and inhibit or alter gene expression via steric hindrance, splicing alterations, initiation of target degradation, or other events.

[0089] In an aspect, ‘'operably linked” means that expression of a gene or a transgene is under the control of a promoter with which it is spatially connected. A promoter can be positioned 5’ (upstream) or 3’ (downstream) of a gene under its control. The distance between the promoter and a gene can be approximately the same as the distance between that promoter and the gene it controls in the gene from which the promoter is derived. As is known in the art. variation in this distance can be accommodated without loss of promoter function.

[0090] In an aspect, “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein must contain at least two amino acids and there is no limitation on the maximum number of amino acids that can comprise a protein’s sequence. The term '‘peptide” can refer to a short chain of amino acids including, for example, natural peptides, recombinant peptides, synthetic peptides, or any combination thereof. Proteins and peptides can include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, and fusion proteins, among others.

[0091] In an aspect, “nucleic acid” or “oligonucleotide” or “polynucleotide” means at least two nucleotides covalently linked together. The depiction of a single strand can also define the sequence of the complementary strand. Thus, a nucleic acid can encompass the complementary strand of a depicted single strand. Many variants of a nucleic acid can be used for the same purpose as a given nucleic acid. Thus, a nucleic acid can encompass substantially identical nucleic acids and complements thereof. A single strand can provide a probe that can hybridize to a target sequence under stringent hybridization conditions. Thus, a nucleic acid can encompass a probe that hybridizes under stringent hybridization conditions. A nucleic acid can be single-stranded, or double-stranded, or can contain portions of both double-stranded and single-stranded sequence. The nucleic acid can be DNA, both genomic and cDNA, RNA, or a hybrid, where the nucleic acid can contain combinations of deoxy ribo- and ribo-nucleotides, and combinations of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine and isoguanine. Nucleic acids can be obtained by chemical synthesis methods or by recombinant methods. In an aspect, the terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid construct,” “nucleotide sequence”, and “polynucleotide'’ can refer to RNA or DNA that is linear or branched, single or double stranded, or a hybrid thereof. The term can encompass RNA / DNA hybrids. When dsRNA is produced synthetically, less common bases, such as inosine, 5-methylcytosine, 6-methyl adenine, hypoxanthine and others can also be used for antisense, dsRNA, and ribozyme pairing. For example, polynucleotides that contain C-5 propyne analogues of uridine and cytidine have been shown to bind RNA with high affinity and to be potent antisense inhibitors of geneexpression. Other modifications, such as modification to the phosphodiester backbone, or the 2’-hydroxy in the ribose sugar group of the RNA can also be made. In an aspect, a “synthetic” nucleic acid or polynucleotide refers to a nucleic acid or polynucleotide that is not found in nature but is constructed by the hand of man and therefore is not a product of nature.

[0092] In an aspect, “polynucleotide” is a sequence of nucleotide bases, and may be RNA. DNA, or DNA- RNA hybrid sequences (including both naturally occurring and non-naturally occurring nucleotides).

[0093] In an aspect, “fragment” or “portion” of a nucleotide sequence can be understood to mean a nucleotide sequence of reduced length relative (e.g., reduced by 1, 2. 3, 4, 5. 6, 7, 8. 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more nucleotides) to a reference nucleic acid or nucleotide sequence and comprising, consisting essentially of, or consisting of a nucleotide sequence of contiguous nucleotides identical or almost identical (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identical) to the reference nucleic acid or nucleotide sequence. Such a nucleic acid fragment or portion according to the disclosure can be, where appropriate, included in a larger polynucleotide of which it is a constituent. In an aspect, a fragment or portion of a nucleotide sequence or nucleic acid sequence can comprise the sequence encoding an exon having one or more mutations or aberrations.

[0094] In an aspect, “fragment” or “portion” of an amino acid sequence can be understood to mean an amino acid sequence of reduced length relative (e.g., reduced by 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, or more amino acids) to a reference amino acid sequence and comprising, consisting essentially of, or consisting of an amino acid sequence of contiguous ammo acids identical or almost identical (e.g., 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identical) to the reference amino acid sequence. Such an amino acid fragment or portion according to the disclosure can be, where appropriate, included in a larger amino acid sequence of which it is a constituent.

[0095] In an aspect, “heterologous” or a “recombinant” nucleotide or amino acid sequence as used interchangeably herein can refer to a nucleotide or an amino acid sequence not naturally associated with a host cell into which it is introduced, including non-naturally occurring multiple copies of a naturally occurring nucleotide or amino acid sequence.

[0096] In an aspect, the term “endogenous” can refer to a gene, protein, compound, or activity that is normally present in a host cell (e.g., a targeted pre-mRNA such as a MYH7 pre-mRNA). In an aspect, an “exogenous” nucleic acid molecule, construct, or sequence (e.g.. an RNAsequence to be trans-spliced) can refer to a nucleic acid molecule or portion of a nucleic acid molecule that is not native to a host cell, but may be homologous to a nucleic acid molecule or portion of a nucleic acid molecule from the host cell.

[0097] In an aspect, "‘promoter” or “promoters” are known to the art. Depending on the level and tissue-specific expression desired, a variety of promoter elements can be used. A promoter can be tissue-specific or ubiquitous and can be constitutive or inducible, depending on the pattern of the gene expression desired. A promoter can be native (endogenous) or foreign (exogenous) and can be a natural or a synthetic sequence. By foreign or exogenous, it is intended that the transcriptional initiation region is not found in the wild-type host into which the transcriptional initiation region is introduced.

[0098] “Tissue-specific promoters” are known to the art and include, but are not limited to, neuron-specific promoters, muscle-specific promoters, liver-specific promoters, skeletal musclespecific promoters, and heart-specific promoters. Ubiquitous / constitutive promoters” are known to the art and include, but are not limited to, a CMV major immediate-early enhancer / chicken beta-actin promoter, a cytomegalovirus (CMV) major immediate-early promoter, an Elongation Factor 1-a (EFl -a) promoter, a simian vacuolating virus 40 (SV40) promoter, an AmpR promoter, a PyK promoter, a human ubiquitin C gene (Ubc) promoter, a MFG promoter, a human beta actin promoter, a CAG promoter, a EGR1 promoter, a FerH promoter, a FerL promoter, a GRP78 promoter, a GRP94 promoter, a HSP70 promoter, a 0-kin promoter, a murine phosphoglycerate kinase (mPGK) or human PGK (hPGK) promoter, a ROSA promoter, human Ubiquitin B promoter, a Rous sarcoma virus promoter, or any other natural or synthetic ubiquitous / constitutive promoters.

[0099] In an aspect, an “inducible promoter” refers to a promoter that can be regulated by positive or negative control. Factors that can regulate an inducible promoter include, but are not limited to, chemical agents (e.g., the metallothionein promoter or a hormone inducible promoter), temperature, and light.

[0100] “Sequence identity” and “sequence similarity” can be determined by alignment of two peptide or two nucleotide sequences using global or local alignment algorithms. Sequences may then be referred to as “substantially identical” or “essentially similar” when they are optimally aligned. For example, sequence similarity or identity can be determined by searching against databases such as FASTA, BLAST, etc., but hits should be retrieved and aligned pairwise to compare sequence identity. Two proteins or two protein domains, or two nucleic acid sequences can have “substantial sequence identity” if the percentage sequence identity is at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99% or more, preferably 90%, 95%, 98%, 99% or more. Such sequences are also referred to as “variants” herein, e.g., other variants of a missing, deficient,and / or mutant protein or enzyme. It should be understood that sequence with substantial sequence identity do not necessarily have the same length and may differ in length. For example, sequences that have the same nucleotide sequence but of which one has additional nucleotides on the 3’-and / or 5'-side are 100% identical.

[0101] In an aspect, “RNA editing” can be a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the structure and function of a protein and may lead to the production of multiple variants of a protein from a single gene.

[0102] In an aspect, insertional and deletional RNA editing can involve the addition and deletion of specific nucleotides or sequences of nucleotides from pre-mRNA. In an aspect, substitutional RNA editing by base modifications is observed in higher eukaryotes, where the base is modified without changing the length of the pre-mRNA.

[0103] In an aspect, “immune tolerance,” “immunological tolerance,” and “immunotolerance” refers to a state of unresponsiveness or blunted response of the immune system to substances (e.g., a disclosed nucleic acid molecule, a disclosed vector, a disclosed transgene product, a disclosed pharmaceutical formulation, a disclosed therapeutic agent, etc.) that have the capacity to elicit an immune response in a subject. Immune tolerance is induced by prior exposure to a specific antigen. Immune tolerance can be determined in a subject by measuring antibodies against a particular antigen or by liver-restricted transgene expression with a viral vector (such as, for example, AAV). Low or absent antibody titers over time is an indicator of immune tolerance. For example, in some embodiments, immune tolerance can be established by having IgG antibody titers of less than or equal to about 12,000, 11,500, 11,000, 10,500, 10,000, 9,500, 9,000, 8,500, 8,000, 7,500, 7,000, 6,500. or 6,000 within following gene therapy (such as the administration of the transgene encoding, for example, a missing, deficient, and / or mutant protein or enzyme).

[0104] In an aspect, the term “package insert” is used to refer to instructions customarily included in commercial packages of therapeutic products, that contain information about the indications, usage, dosage, administration, contraindications and / or warnings concerning the use of such therapeutic products.

[0105] In an aspect, the term “in combination” in the context of the administration of other therapies (e.g., other agents) includes the use of more than one therapy (e.g., drug therapy). Administration “in combination with” one or more further therapeutic agents includes simultaneous (e.g., concurrent) and consecutive administration in any order. The use of the term■‘in combination’’ does not restrict the order in which therapies are administered to a subject. By way of non-limiting example, a first therapy (e.g., a disclosed nucleic acid molecule, a disclosed vector, a disclosed pharmaceutical formulation, or a combination thereof) may be administered prior to (e.g.. 1 minute, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours. 8 hours, 12 hours. 24 hours, 48 hours. 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks), concurrently, or after (e.g., 1 minute, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours. 7 hours, 8 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours. 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks. 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, or 12 weeks or longer) the administration of a second therapy (e.g., agent) to a subject having or diagnosed with a disease or disorder (such as a genetic disease or disorder such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations).

[0106] Disclosed are the components to be used to prepare the disclosed nucleic acid molecules, disclosed vectors, or disclosed pharmaceutical formulations as well as the disclosed nucleic acid molecules, disclosed vectors, or disclosed pharmaceutical formulations used within the methods disclosed herein. These and other materials are disclosed herein, and it is understood that when combinations, subsets, interactions, groups, etc. of these materials are disclosed that while specific reference of each various individual and collective combinations and permutation of these compounds cannot be explicitly disclosed, each is specifically contemplated and described herein. For example, if a particular compound is disclosed and discussed and a number of modifications that can be made to a number of molecules including the compounds are discussed, specifically contemplated is each and every combination and permutation of the compound and the modifications that are possible unless specifically indicated to the contrary. Thus, if a class of molecules A, B, and C are disclosed as well as a class of molecules D, E, and F and an example of a combination molecule, A-D is disclosed, then even if each is not individually recited each is individually and collectively contemplated meaning combinations, A-E, A-F, B-D, B-E, B-F, C-D, C-E, and C-F are considered disclosed. Likewise, any subset or combination of these is also disclosed. Thus, for example, the sub-group of A-E, B-F, and C-E would be considered disclosed. This concept applies to all aspects of this application including, but not limited to, steps in methods of making and using the compositions of the invention. Thus, if there are a variety of additional steps that can be performed it is understood that each of these additional steps can be performed with any specific embodiment or combination of embodiments of the methods of the invention.B. Compositions for Transcriptome Engineering1. Myosin Heavy Chain (MYH7)

[0107] MYH7 can be identified by the following accession numbers: HGNC (7577); NCBI Gene (4625); Ensembl (ENSG00000092054); OMIM (160760); and UniProtKB / Swiss-Prot (P12883). MYH7 can be identified by accession numbers: NG 007884.1 (genomic DNA - SEQ ID NO:110), NM_000257.4 (mRNA - SEQ ID NO:111), or NP_000248.2 (protein - SEQ ID NO: 112).2. Hypertrophic Cardiomyopathy

[0108] Hypertrophic cardiomyopathy is a heart condition characterized by thickening (hypertrophy) of the heart (cardiac) muscle. When multiple members of a family have the condition, it is known as familial hypertrophic cardiomyopathy. Hypertrophic cardiomyopathy also occurs in people with no family history; these cases are considered nonfamilial hypertrophic cardiomyopathy.

[0109] In familial hypertrophic cardiomyopathy, cardiac thickening usually occurs in the interventricular septum, which is the muscular wall that separates the lower left chamber of the heart (the left ventricle) from the lower right chamber (the right ventricle). In some people, thickening of the interventricular septum impedes the flow of oxygen-rich blood from the heart, which may lead to an abnormal heart sound during a heartbeat (heart murmur) and other signs and symptoms of the condition. Other affected individuals do not have physical obstruction of blood flow, but the pumping of blood is less efficient, which can also lead to symptoms of the condition. Familial hypertrophic cardiomyopathy often begins in adolescence or young adulthood, although it can develop at any time throughout life.

[0110] The symptoms of familial hypertrophic cardiomyopathy are variable, even within the same family. Many affected individuals have no symptoms. Other people with familial hypertrophic cardiomyopathy may experience chest pain; shortness of breath, especially with physical exertion; a sensation of fluttering or pounding in the chest (palpitations); lightheadedness; dizziness; and fainting. While most people with familial hypertrophic cardiomyopathy are symptom-free or have only mild symptoms, this condition can have serious consequences. It can cause abnormal heart rhythms (arrhythmias) that may be life threatening. People with familial hypertrophic cardiomyopathy have an increased risk of sudden death, even if they have no other symptoms of the condition. A small number of affected individuals develop potentially fatal heart failure, which may require heart transplantation.3. Nucleic Acid Molecules5’ Replacement Constructs

[0111] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be transspliced to a targeted endogenous pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.

[0112] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous pre-mRNA.

[0113] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA.

[0114] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.

[0115] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linkedto an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous pre-mRNA.

[0116] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous pre-mRNA.

[0117] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0118] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to betrans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0119] Disclosed herein is a nucleic acid molecule an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0120] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0121] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter an exogenous RNA to be trans-spliced to a targeted endogenous premRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be transspliced comprises one or more exons located in the 5’ portion of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0122] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0123] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.

[0124] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA.

[0125] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be transspliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA.

[0126] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.

[0127] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA.

[0128] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences,wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA.

[0129] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be transspliced to a targeted endogenous MYH7 pre-mRNA; a 5' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0130] Disclosed herein is a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0131] Disclosed herein is a nucleic acid molecule an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half ofthe open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0132] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0133] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0134] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targetedendogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0135] In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more pathogenic mutations or aberrations. In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more mutations or aberrations. In an aspect, Table 6 provides a listing of mutations or aberrations in MYH7.

[0136] an aspect, a disclosed nucleic acid molecule can lack a CRISPR-associated protein or can be CRISPR-free. In an aspect, a disclosed multivalent guide trans-splicing RNA molecule can lack a CRISPR-associated protein or can be CRISPR-free.

[0137] In an aspect, one or more disclosed RNA structures can improve and / or can enhance trans-splicing efficiency. In an aspect, one or more disclosed RNA structures can stabilize the pre-mRNA. In an aspect, one or more disclosed RNA structures can localize the RNA to the nucleus. In an aspect, one or more disclosed RNA structures can stabilize the interaction between the targeted endogenous pre-mRNA molecule and the exogenous RNA to be trans-spliced.

[0138] In an aspect of a disclosed 5' replacement construct, a disclosed RNA structure can comprise the sequence of SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, or any combination thereof. In an aspect of a disclosed 5’ replacement construct, a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106 or any combination thereof. In an aspect of a disclosed 5' replacement construct, a disclosed RNA structure can comprise the sequence of SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, or a fragment thereof, or any combination thereof. In an aspect of a disclosed 5’ replacement construct, a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106, or a fragment thereof, or any combination thereof.

[0139] In an aspect of a disclosed 5’ replacement construct, a disclosed RNA structure can comprise an RNA structure identified in Table 1. In an aspect of a disclosed 5’ replacement construct, a disclosed RNA structure can comprise a combination of one or more RNA structures identified in Table 1.Table 1 - List of RNA Structures

[0140] In an aspect, a disclosed guide RNA sequence can bind to the 5’ end of the targeted endogenous MYH7 pre-mRNA. In an aspect, a disclosed 3’ portion of the targeted endogenous MYH7 pre-mRNA can be trans-spliced with the exogenous RNA. In an aspect, a disclosed RNA guide sequence can be specific for an endogenous MYH7 pre-mRNA having one or more mutations. In an aspect, a disclosed RNA guide sequence can be specific for an endogenous MYH7 pre-mRNA having one or more exonic mutations.

[0141] In an aspect, a disclosed RNA guide sequence can be specific for an endogenous MYH7 pre-mRNA having one or more intromc mutations. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29. or fragment thereof, or any combination thereof. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can comprise a sequence having at least 80% identity, at least 85% identity, at least 90% identity’, at least 95% identity, or more than 95% identity to the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29, orfragment thereof, or any combination thereof. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof. In an aspect of a disclosed 5' replacement construct, a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:30 - SEQ ID NO:43. or a fragment thereof, or any combination thereof. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can comprise a sequence having at least 80% identity, at least 85% identity, at least 90% identity7, at least 95% identity, or more than 95% identity to the sequence of any one of SEQ ID NO: 30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0142] In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can target the intron between exon 22 and exon 23 of MYH7. In an aspect of a disclosed 5’ replacement construct, one or more disclosed guide RNA sequences can target the intron between exon 22 and exon 23 of MYH7. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can comprise a guide RNA sequence identified in Table 2. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can comprise a combination of one or more guide RNA sequences identified in Table 2.Table 2 - List of Exemplary Guide RNA Sequences

[0143] In an aspect, Guide 11, 16, and 25 can independently enable trans-splicing. In an aspect, combinations of Guide 11, 16, and 25 can enable trans-splicing. In an aspect, the combination of Guide 16 and Guide 25 can significantly enhance trans-splicing efficiency. In an aspect, the combination of Guide 16 and Guide 25 with the combination of SLBP and TAR can significantly enhance trans-splicing efficiency.

[0144] In an aspect, a disclosed guide RNA sequence can be extended by one or more bp. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can be extended by 20 bp, 30 bp, 40 bp, 50 bp, 60 bp, 70 bp, 80 bp, 90 bp, 100 bp, 110 bp, 120 bp, 130 bp, 140 bp, 150 bp. or more than 150 bp. In an aspect, a disclosed guide RNA sequence can be truncated by one or more bp. In an aspect of a disclosed 5’ replacement construct, a disclosed guide RNA sequence can be truncated by 1 bp, 2 bp, 3 bp, 4 bp, 5 bp, 6 bp, 7 bp, 8 bp, 9 bp, 10 bp, 11 bp, 12 bp, 13 bp, 14 bp, 15 bp, 16 bp, 17 bp, 18 bp, 19 bp, 20 bp, 21 bp, 22 bp, 23 bp, 24 bp, 25 bp, 26 bp, 27 bp, 28 bp, 29 bp, 30 bp, or more than 30 bp. In an aspect, a disclosed guide RNA sequence can be a guide RNA sequence listed in Table 3.Table 3 - List of Exemplary Truncated and / or Extended Guide RNA Sequences

[0145] In an aspect of a disclosed 5’ replacement construct a disclosed RNA structure can comprise the sequence of SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, or any combination thereof, and a disclosed guide RNA sequence can comprise the sequence of (i) any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0146] In an aspect of a disclosed 5’ replacement construct, a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106 or any combination thereof, and a disclosed guide RNA sequence can comprise the sequence of (i) any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0147] In an aspect of a disclosed 5’ replacement construct, a disclosed RNA structure can comprise the sequence of SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, or a fragment thereof, or any combination thereof, and a disclosed guide RNA sequence can comprise the sequence of (i) any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0148] In an aspect of a disclosed 5’ replacement construct, a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106, or a fragment thereof, or any combination thereof, and a disclosed guide RNA sequence can comprise the sequence of (i) any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0149] In an aspect of a disclosed 5’ replacement construct, one or more disclosed RNA structures can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO: 99 or any combinationthereof, and one or more disclosed guide RNA sequences can comprise the sequence of (i) one or more of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) one or more of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) one or more of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) one or more of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0150] In an aspect of a disclosed 5 ’ replacement construct, one or more disclosed RNA structures can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106 or any combination thereof, and one or more disclosed guide RNA sequences can comprise the sequence of (i) one or more of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) one or more of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) one or more of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) one or more of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0151] In an aspect of a disclosed 5’ replacement construct, one or more disclosed RNA structures can comprise the sequence of any one of SEQ ID NO: 97 - SEQ ID NO: 99 or a fragment thereof, or any combination thereof, and one or more disclosed guide RNA sequences can comprise the sequence of (i) one or more of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) one or more of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) one or more of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) one or more of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof. In an aspect of a disclosed 5’ replacement construct, one or more disclosed RNA structures can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106, or a fragment thereof, or any combination thereof, and one or more disclosed guide RNA sequences can comprise the sequence of (i) one or more of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) one or more of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) one or more of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) one or more of SEQ ID NO:30 - SEQ ID NO:43. or a fragment thereof, or any combination thereof.

[0152] In an aspect of a disclosed 5’ replacement construct, a disclosed combination of a disclosed RNA structure and a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:44 - SEQ ID NO:47. In an aspect of a disclosed 5’ replacement construct, a disclosed combination can comprise a disclosed RNA structure followed by a disclosed guide RNA sequence. In an aspect of a disclosed 5’ replacement construct, a disclosed combination can comprise a disclosed guide RNA sequence followed by a disclosed RNA structure.

[0153] In an aspect of a disclosed 5 ’ replacement construct, a disclosed combination can comprise a disclosed RNA structure followed by a disclosed guide RNA sequence, wherein a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQID NO: 100 - SEQ ID NO: 106, and wherein a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43. In an aspect of a disclosed 5' replacement construct, a disclosed combination can comprise a disclosed RNA structure followed by a disclosed guide RNA sequence, wherein a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment thereof, and wherein a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 -SEQ ID NO:43 or a fragment thereof. In an aspect of a disclosed 5‘ replacement construct, a disclosed combination can comprise a disclosed guide RNA sequence followed by a disclosed RNA structure, wherein a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43, and wherein a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106. In an aspect of a disclosed 5‘ replacement construct, a disclosed combination can comprise a disclosed guide RNA sequence followed by a disclosed RNA structure, wherein a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 or a fragment thereof, and wherein a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment.

[0154] In an aspect of a disclosed 5’ replacement construct, the combination of two disclosed RNA structures and two disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:48 - SEQ ID NO:60. In an aspect of a disclosed 5‘ replacement construct, the combination of two disclosed RNA structures and two disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:48 - SEQ ID NO: 60 or a fragment thereof. In an aspect of a disclosed 5’ replacement construct, the combination of two disclosed RNA structures and two disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:48 - SEQ ID NO:60, wherein the disclosed RNA structures can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106. and the disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43. In an aspect, a disclosed combination of two disclosed RNA structures and two disclosed RNA guide sequences, the disclosed RNA structures can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 -SEQ ID NO: 106 or a fragment, and the disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 or a fragment thereof.

[0155] In an aspect a disclosed combination of disclosed RNA structures and disclosed RNA guide sequences can comprise the sequence set forth in Table 4.Table 4 - List of Exemplary Combinations of RNA Structures and RNA Guides

[0156] In an aspect of a disclosed 5’ replacement construct disclosed exogenous RNA to be transspliced can comprise the sequence set forth in SEQ ID NO:95. In an aspect of a disclosed 5’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO:95 and be optimized by addition of one or more disclosed RNA structures and one or more disclosed guide RNA sequences. In an aspect of a disclosed 5’ replacement construct a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO:95 and can be combined with one or more disclosed RNA structures and one or more disclosed guide RNA sequences. In an aspect of a disclosed 5’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO: 95 and can be combined with one or more disclosed RNA structures and one or more disclosed guide RNA sequences, wherein the disclosed RNA structures can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment, andwherein the disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 or a fragment thereof. In an aspect of a disclosed 5’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:95. In an aspect of a disclosed 5’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:95 and can be combined with one or more disclosed RNA structures and one or more disclosed guide RNA sequences.

[0157] In an aspect of a disclosed 5’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:95 and can be combined with one or more disclosed RNA structures and one or more disclosed guide RNA sequences, wherein the disclosed RNA structures can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment, and wherein the disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:01 -SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 or a fragment thereof.

[0158] In an aspect, Guide 5, 13, and 19 can independently enable trans-splicing. In an aspect, combinations of Guide 5, 13, and 19 can enable trans-splicing. In an aspect, the combination of Guide 5 and Guide 19 can significantly enhance trans-splicing efficiency. In an aspect, the combination of Guide 5 and Guide 13 can significantly enhance trans-splicing efficiency. In an aspect, the combination of Guide 13 and Guide 19 can significantly enhance trans-splicing efficiency. In an aspect, the combination of Guides 5 and 19 with the combination of SLBP, MALAT1, TAR, or a combination thereof can significantly enhance trans-splicing efficiency. In an aspect, the combination of Guides 5 and 13 with the combination of SLBP, MALAT1, TAR, or a combination thereof can significantly enhance trans-splicing efficiency. In an aspect, the combination of Guides 13 and 19 with the combination of SLBP, MALAT1, TAR, or a combination thereof can significantly enhance trans-splicing efficiency.

[0159] In an aspect, a first disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 can be combined with a second disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43. In an aspect, a first disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 can be combined with a second disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43. and can further be combined with a disclosed RNA structure of any one of SEQ IDNO:97- SEQ ID NO: 106. In an aspect, a first disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 can be combined with a second disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO: 43, and can further be combined with a first disclosed RNA structure of any one of SEQ ID NO:97- SEQ ID NO: 106 and a second disclosed RNA structure of any one of SEQ ID NO:97 - SEQ ID NO: 106.

[0160] In an aspect of a disclosed 5’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO: 89. In an aspect of a disclosed 5' replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:89.

[0011] In an aspect, Table 5 provides a listing of exemplary 5’ replacement constructs.Table 5 - List of Exemplary 5’ Replacement Constructs

[0162] In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more mutations or aberrations (such as, for example, those listed in Table 6). In an aspect of a disclosed targeted endogenous MYH7 pre-mRNA, one or more disclosed mutations or aberrations can be in the 5’ portion of the pre-mRNA. In an aspect, one or more disclosed mutations or aberrations in one or more exons can contribute to pathogenesis of one or more cells. In an aspect, one or more disclosed mutations or aberrations can inhibit translation of the encoded protein. In an aspect, one or more disclosed mutations or aberrations can modify translation of the encoded protein. In an aspect, one or more disclosed mutations or aberrations can generate an encoded protein having a non-sense mutation or a missense mutation.

[0163] In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more mutations in one or more exons. In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more mutations in one or more introns. In an aspect, one or more disclosed exonic mutations can contribute to pathogenesis in one or more cells. In an aspect, one or more disclosed intronic mutations can contribute to pathogenesis in one or more cells. In an aspect, a disclosed resulting chimeric RNA transcript can comprise the 3’ portion of the targeted endogenous pre-mRNA and the 5’ portion of the exogenous RNA. In an aspect, a disclosedtargeted endogenous pre-mRNA and a disclosed exogenous RNA can encode the same protein coding gene (i.e., MYH7). In an aspect, a disclosed targeted endogenous pre-mRNA and a disclosed exogenous RNA can comprise one or more exons of the same protein coding gene (i.e., MYH7).

[0164] In an aspect, a disclosed 5’ replacement construct can redress one or more of the 165 mutations identified below in Table 6.Table 6 - List of Mutations in MYH7> > > > > > > > > > > > > > > > > > > > > > > >> > > > > > > > > > > > > > > > > > > > > > > > > > > > >> > > > > > > > > > > > > > > > > > > > > > > > > > > > > >> > > > > > > > > > > > > > > > > > > > > > > > > > > > > >> > > > > > > > > > > > > > > > > > > > > > > > > > > >>> > > >> >> > > > > > >

[0165] In an aspect a disclosed 5’ hemi intron can comprise the sequence set forth in SEQ ID NO:91. In an aspect, a disclosed 5’ hemi intron can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO 91.

[0166] In an aspect, a disclosed 5’ hemi intron can be linked to a disclosed exogenous RNA to be trans-spliced. In an aspect, a disclosed 5’ hemi intron can comprise a splice donor sequence that can recruit a U1 snRNP. In an aspect, a disclosed 5’ hemi intron can comprise one or more U1 binding sites. In an aspect, a disclosed 5’ hemi intron can comprise at least 1, at least 2, at least 3, at least 4, or at least 5 U1 binding sites. In an aspect, a disclosed 5’ hemi intron comprising one or more U1 binding sites can comprise the sequence set forth in SEQ ID NO:92. In an aspect, a disclosed 5’ hemi intron comprising one or more U1 binding sites can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or more than 95% identity to the sequence set forth in SEQ ID NO:92. In an aspect, a disclosed U1 snRNP can enable formation of a spliceosome with a disclosed exogenous RNA to be trans-spliced and a disclosed targeted endogenous MYH7 pre-mRNA.

[0167] In an aspect, a disclosed 5’ hemi intron can be recognized by the spliceosome in a host cell. In an aspect, a disclosed 5’ hemi intron can be recognized by nuclear splicing components in a host cell. In an aspect, a disclosed exogenous RNA can induce a trans-splicing event. In an aspect, a disclosed 5 ' hemi intron can be recognized by nuclear splicing components within a host cell. In an aspect, proper assembly of a spliceosome between a disclosed exogenous RNA to be trans-spliced and a disclosed targeted endogenous MYH7 pre-mRNA can facilitate the joining of the 5’ end of the disclosed targeted endogenous MYH7 pre-mRNA with a disclosed exogenous RNA to be trans-spliced. In an aspect, proper assembly of a spliceosome between a disclosed exogenous RNA to be trans-spliced and a disclosed targeted endogenous MYH7 pre-mRNA can facilitate the joining of exons 23-40 of the disclosed targeted endogenous MYH7 pre-mRNA with exons 1-22 of the disclosed exogenous RNA to be trans-spliced.

[0168] In an aspect, proper assembly of a spliceosome between a disclosed exogenous RNA to be trans-spliced and a disclosed targeted endogenous MYH7 pre-mRNA can facilitate the joining of exons 23-40 of the disclosed targeted endogenous MYH7 pre-mRNA with exons 1-22 of the disclosed exogenous RNA to be trans-spliced, thereby correcting and / or replacing one or more pathogenic mutations or aberrations in exons 1-22 of the targeted endogenous MYH7 pre-mRNA.

[0169] In an aspect, a disclosed 5’ hemi intron can facilitate the trans-splicing of the exogenous RNA to the exon immediately 5 ’ to the targeted intron in the endogenous pre-mRNA. In an aspect, a disclosed 5’ replacement construct can enable the 5’ replacement of the targeted endogenous pre-MYH7 mRNA having one or more mutations or aberrations. In an aspect, a disclosed 5’ replacement construct can enable the replacement of exons 1-22 of the targeted endogenous MYH7 pre-mRNA.

[0170] In an aspect, a disclosed 5’ replacement construct can be used in a trans-splicing event. In an aspect, a disclosed 5’ replacement construct can be used in conjunction with one or more blocking U7 snRNAs. In an aspect, a disclosed 5’ replacement construct can further comprise the sequence of one or more blocking U7 snRNAs. In an aspect, a disclosed blocking U7 snRNA can be used to inhibit or decrease cis-splicing during a trans-splicing event comprising a disclosed 5’ replacement construct. In aspect, when used with one or more disclosed blocking U7 snRNAs, a disclosed 5’ replacement construct can more efficiently drive trans-splicing. In an aspect, a disclosed blocking U7 snRNA can comprise the sequence set forth in any one of SEQ ID NO:62 - SEQ ID NO:71. In an aspect, a disclosed blocking U7 snRNA can comprise a sequence having at least 90% identity to the sequence set forth in any one of SEQ ID NO:62 - SEQ ID NO:71. In an aspect, a disclosed U7 snRNA of SEQ ID NO:62 can be used with a disclosed guide RNA sequence of SEQ ID NO:01. In an aspect, a disclosed U7 snRNA of SEQ ID NO:63 can be used with a disclosed guide RNA sequence of SEQ ID NO:02. In an aspect, a disclosed U7 snRNA ofSEQ ID NO: 64 can be used with a disclosed guide RNA sequence of SEQ ID NO: 03. In an aspect, a disclosed U7 snRNA of SEQ ID NO:65 can be used with a disclosed guide RNA sequence of SEQ ID NO:04. In an aspect, a disclosed U7 snRNA of SEQ ID NO:66 can be used with a disclosed guide RNA sequence of SEQ ID NO:05. In an aspect, a disclosed U7 snRNA of SEQ ID NO:67 can be used with a disclosed guide RNA sequence of SEQ ID NO:06. In an aspect, a disclosed U7 snRNA of SEQ IDNO:68 can be used with a disclosed guide RNA sequence of SEQ ID NO:07. In an aspect, a disclosed U7 snRNA of SEQ ID NO:69 can be used with a disclosed guide RNA sequence of SEQ ID NO:08. In an aspect, a disclosed U7 snRNA of SEQ ID NO:70 can be used with a disclosed guide RNA sequence of SEQ ID NO:09. In an aspect, a disclosed U7 snRNA of SEQ ID NO:71 can be used with a disclosed guide RNA sequence of SEQ ID NO:10.

[0171] In an aspect, expression of a disclosed U7 snRNA can be driven by a U7 promoter. In an aspect, a U7 snRNA expression construct can comprise a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator.

[0172] In an aspect, a disclosed U7 snRNA expression construct comprising a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator can comprise the sequence set forth in any one of SEQ ID NO: 73 - SEQ ID NO: 82. In an aspect, a disclosed U7 snRNA expression construct comprising a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator can comprise a sequence having at least 90% to the sequence set forth in any one of SEQ ID NO: 73 - SEQ ID NO: 82.

[0173] In an aspect, a disclosed U7 snRNA expression construct comprising a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator can comprise the sequence set forth in any one of SEQ ID NO:73 - SEQ ID NO:82, and can be used in conjunction with one or more 5’ replacement constructs.

[0174] In an aspect, a disclosed U7 snRNA expression construct comprising a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator can comprise a sequence having at least 90% to the sequence set forth in any one of SEQ ID NO: 73 - SEQ ID NO:82, and can be used in conjunction with one or more 5’ replacement constructs.

[0175] In an aspect, a disclosed U7 snRNA expression construct can be incorporated into a viral vector or a non-viral vector. In an aspect, a disclosed U7 snRNA expression construct can be incorporated into a viral vector or a non-viral vector comprising a disclosed nucleic acid molecule.

[0176] In an aspect, a disclosed promoter for the 5’ replacement construct can be tissue-specific or ubiquitous and can be constitutive or inducible, depending on the pattern of the expression desired. In an aspect, a disclosed promoter can be native or foreign and can be a natural or a synthetic sequence. By foreign, it is intended that the transcriptional initiation region is not foundin the w ild-type host into which the transcriptional initiation region is introduced. In an aspect, a disclosed promoter can be a promoter / enhancer. In an aspect, a disclosed promoter for the disclosed nucleic acid molecule can be an endogenous promoter. In an aspect, a disclosed endogenous promoter can be an endogenous promoter / enhancer. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can generally be obtained from a non-coding region upstream of a transcription initiation site of a gene of interest. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can be used for constitutive and efficient expression of a disclosed gene. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be a CMV promoter or a CMV promoter / enhancer. In an aspect, a disclosed CMV protomer can comprise the sequence set forth in SEQ ID NO:83. CMV promoters and CMV promoters / enhancers are well known to the art. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be any eukaryotic RNA polymerase II promoter.

[0177] In an aspect, a disclosed nucleic acid molecule can further comprise can further comprise a UTR. In an aspect, a disclosed nucleic acid molecule can further comprise a polyadenylation sequence. In an aspect, a disclosed nucleic acid molecule can further comprise a sequence for a promoter. In an aspect, a disclosed nucleic acid molecule can further comprise a spacer region. In an aspect, a disclosed spacer region can separate the 5' splice region from the one or more RNA structures. In an aspect, a disclosed spacer region can comprise any known spacer. In an aspect, a disclosed spacer region can comprise a consensus splicing motif (e.g., such as U1 or U2). In an aspect, a disclosed spacer region can comprise a limited number of consensus splicing motifs (e.g., such as U1 or U2).

[0178] In an aspect, a disclosed nucleic acid molecule can further comprise one or more nuclear localization signals (NLS). NLS are known to the skilled person in the art. In an aspect, a disclosed NLS can comprise any NLS known to the art. As known to the art (see, e.g., Lu J, et al. (2021) Cell Commun Signal. 19:60, which is incorporated herein by reference for its teachings of NLS), nuclear localization signals (NLS) are generally short peptides that act as a signal fragment that mediates the transport of proteins from the cytoplasm into the nucleus.

[0179] In an aspect, a disclosed nucleic acid molecule can further comprise one or more nuclear retention elements (NRE). NRE are known to the skilled person in the art. In an aspect, a disclosed NRE can comprise SIRLOIN (SEQ ID NO: 113) or BORG (SEQ ID NO: 114).

[0180] In an aspect, a disclosed nucleic acid molecule can further comprise one or more Flavivirus genetic elements. In an aspect, Flavivirus genetic elements can comprise one or more Flavivirus 3’ untranslated region (3' UTR), one or more subgenomic Flavivirus RNA (siRNA) elements, one or more Flavivirus XRN1 -resistant RNA (xrRNA) elements, one or more Flavivirus dumbbell(DB) RNA elements, one or more Flavivirus 3’ stem loop (3’ SL) elements, or any combination thereof. (See WO 2022 / 182835 for a description of Flavivirus gene elements).

[0181] In an aspect, a disclosed cell or a disclosed host cell can be in a subject. In an aspect, a subject can be a human patient and can be male or female. In an aspect, a subject can have a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations). In an aspect, a subject can be treatment-naive. In an aspect, a patient can refer to a subject afflicted with a disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations). In an aspect, a subject or a patient can be seeking treatment or receiving treatment for a disease or disorder (i.e., hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations).

[0182] In an aspect, a disclosed nucleic acid molecule comprising a 5’ replacement construct can be packaged into lipid nanoparticles. In an aspect, a disclosed nucleic acid molecule comprising a 5’ replacement construct can be packaged into a viral vector. In an aspect, a disclosed viral vector can comprise an AAV vector. In an aspect, a disclosed nucleic acid molecule can be packaged into a non-viral carrier. In an aspect, a disclosed nucleic acid molecule can be incorporated into a plasmid. In an aspect, a disclosed nucleic acid molecule can be incorporated into lipid nanoparticles.

[0183] Disclosed herein is an expression cassette comprising disclosed 5' replacement construct. Disclosed herein is an expression cassette comprising a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is an expression cassette comprising a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.

[0184] Disclosed herein is an expression cassette comprising (1) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (2) a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (3) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’portion of the targeted endogenous MYH7 pre-mRNA; (4) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (5) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (6) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (7) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (8) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (9) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (10) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ‘ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (11) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (12) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be transspliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (13) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (14) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (15) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (16) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (17) anucleic acid molecule an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (18) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targetedendogenous MYH7 pre-mRNA; (19) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (20) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (21) a multivalent guide trans-splicing RNA molecule comprising a promoter an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (22) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; or (23) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0185] In an aspect, use of a disclosed 5’ replacement construct or a disclosed expression cassette comprising a disclosed 5’ replacement construct can be used to restore and / or return MYH7 expression to a wild-type, normal, or control expression level. In an aspect, a disclosed nucleic acid molecule can restore one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation. In an aspect, a disclosed nucleic acid molecule can restore the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme (i.e., MYH7). In an aspect, restoring one or more aspects of cellular homeostasis and / or cellular functionality can comprise one or more of the following: (i) correcting cell starvation in one or more cell types; (ii) normalizing aspects of the autophagy pathway (such as, for example, correcting, preventing, reducing, and / or ameliorating autophagy); (iii) improving, enhancing, restoring, and / or preserving mitochondrial functionality and / or structural integrity; (iv) improving, enhancing, restoring, and / or preserving organelle functionality and / or structural integrity; (v) correcting enzyme dysregulation; (vi) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of the multi-systemic manifestations of a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations); (vii) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations); or (viii) any combination thereof. In an aspect, restoring one or more aspects of cellular homeostasis can comprise improving, enhancing, restoring, and / or preserving one or more aspects of cellular structural and / or functional integrity.

[0186] In an aspect, restoring the activity and / or functionality' of a missing, deficient, and / or mutant protein or enzyme (i.e., MYH7) can comprise a 10%, 20%. 30%. 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount of restoration when compared to a pre-existing level such as, for example, a pre-treatment level. In an aspect, the amount of restoration can be 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% more than a pre-existing level such as, for example, a pre-treatment level. In an aspect, restoration can be measured against a control level or a reference level (e.g., determined, for example, using one or more subjects not having a missing, deficient, and / or mutant protein or enzyme like MYH7). In an aspect, restoration can be a partial or incomplete restoration. In an aspect, restoration can be complete or near complete restoration such that the level of expression, activity', and / or functionality' is similar to that of a wild-type or control level.

[0187] In an aspect, a disclosed nucleic acid sequence can further comprising the sequence of one or more disclosed blocking U7 snRNAs. In an aspect, a disclosed 5’ replacement construct can further comprise the sequence of one or more disclosed blocking U7 snRNAs. In an aspect, adisclosed nucleic acid sequence comprising a 5’ replacement construct can further comprise the sequence of one or more disclosed blocking U7 snRNAs.3’ Replacement Constructs

[0188] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA.

[0189] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA.

[0190] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3‘ portion of the targeted endogenous pre-mRNA.

[0191] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3' portion of the targeted endogenous pre-mRNA.

[0192] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA.

[0193] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA.

[0194] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; a guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA.

[0195] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linkedto an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA.

[0196] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous pre-mRNA.

[0197] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3 ’ portion of the targeted endogenous pre-mRNA.

[0198] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA.

[0199] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA.

[0200] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0201] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0202] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein theexogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0203] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3 ’ portion of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0204] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0205] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0206] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0207] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0208] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located inthe 3’ portion of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be transspliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0209] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0210] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0211] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA.

[0212] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA.

[0213] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3 ’ portion of the targeted endogenous MYH7 pre-mRNA.

[0214] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA.

[0215] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; a guide RNA sequences; a 3' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA.

[0216] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA.

[0217] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 premRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA.

[0218] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA.

[0219] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0220] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0221] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0222] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0223] Disclosed herein is a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame ofthe targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0224] Disclosed herein is a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0225] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0226] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0227] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3 ’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0228] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3' portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0229] Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ' hemi intron linked to an exogenousRNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0230] In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more pathogenic mutations or aberrations. In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more mutations or aberrations. In an aspect, Table 6 provides a listing of mutations or aberrations in MYH7.

[0231] an aspect, a disclosed nucleic acid molecule can lack a CRISPR-associated protein or can be CRISPR-free. In an aspect, a disclosed multivalent guide trans-splicing RNA molecule can lack a CRISPR-associated protein or can be CRISPR-free.

[0232] In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise the sequence of SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, or any combination thereof. In an aspect of a disclosed 3‘ replacement construct, a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106 or any combination thereof. In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise the sequence of SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, or a fragment thereof, or any combination thereof. In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106, or a fragment thereof, or any combination thereof.

[0233] In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise an RNA structure identified in Table 1. In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise a combination of one or more RNA structures identified in Table 1.

[0234] In an aspect, a disclosed guide RNA sequence can bind to the 3’ end of the targeted endogenous MYH7 pre-mRNA. In an aspect, a disclosed 5’ portion of the targeted endogenous MYH7 pre-mRNA can be trans-spliced with the exogenous RNA.

[0235] In an aspect, a disclosed RNA guide sequence can be specific for an endogenous MYH7 pre-mRNA having one or more mutations or aberrations. In an aspect, a disclosed RNA guide sequence can be specific for an endogenous MYH7 pre-mRNA having one or more exonic mutations or aberrations. In an aspect, a disclosed RNA guide sequence can be specific for an endogenous MYH7 pre-mRNA having one or more intronic mutations or aberrations.

[0236] In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can comprise the sequence of any one of SEQ IDNO:01 - SEQ ID NO:29, or any combination thereof. In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof. In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can comprise a sequence having at least 80% identity, at least 85% identity, at least 90% identity7, at least 95% identity, or more than 95% identity to the sequence of any one of SEQ ID NO: 01 - SEQ ID NO:29, or fragment thereof, or any combination thereof. In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof. In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof. In an aspect of a disclosed 3' replacement construct, a disclosed guide RNA sequence can comprise a sequence having at least 80% identity, at least 85% identity, at least 90% identity7, at least 95% identity, or more than 95% identity7to the sequence of any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0237] In an aspect, Guide 11. 16. and 25 can independently enable trans-splicing. In an aspect, combinations of Guide 11, 16, and 25 can enable trans-splicing. In an aspect, the combination of Guide 16 and Guide 25 can significantly enhance trans-splicing efficiency. In an aspect, the combination of Guide 16 and Guide 25 with the combination of SLBP and TAR can significantly enhance trans-splicing efficiency.

[0238] In an aspect, a first disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 can be combined with a second disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43.

[0239] In an aspect, a first disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 can be combined with a second disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43, and can further be combined with a disclosed RNA structure of any one of SEQ ID NO:97- SEQ ID NO: 106. In an aspect, a first disclosed guide RNA sequence of any one of SEQID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 can be combined with a second disclosed guide RNA sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO: 43, and can further be combined with a first disclosed RNA structure of any one of SEQ ID NO:97- SEQ ID NO: 106 and a second disclosed RNA structure of any one of SEQ ID NO:97 - SEQ ID NO: 106.

[0240] In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can be extended by 20 bp, 30 bp, 40 bp, 50 bp, 60 bp, 70 bp, 80 bp, 90 bp, 100 bp, 110 bp, 120 bp, 130 bp, 140 bp, 150 bp, or more than 150 bp. In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can be truncated by 1 bp, 2 bp. 3 bp. 4 bp, 5 bp, 6 bp, 7 bp, 8 bp, 9 bp, 10 bp, 11 bp, 12 bp, 13 bp, 14 bp, 15 bp, 16 bp, 17 bp, 18 bp, 19 bp, 20 bp, 21 bp, 22 bp, 23 bp, 24 bp, 25 bp, 26 bp, 27 bp, 28 bp, 29 bp, 30 bp, or more than 30 bp.

[0241] In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can target the intron between exon 22 and exon 23 of MYH7. In an aspect of a disclosed 3’ replacement construct, one or more disclosed guide RNA sequences can target the intron between exon 22 and exon 23 of MYEI7.

[0242] In an aspect of a disclosed 3’ replacement construct, a disclosed guide RNA sequence can comprise a guide RNA sequence identified in Table 2. In an aspect of a disclosed 3 ’ replacement construct, a disclosed guide RNA sequence can comprise a combination of one or more guide RNA sequences identified in Table 2.

[0243] In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise the sequence of SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, or any combination thereof, and a disclosed guide RNA sequence can comprise the sequence of (i) any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0244] In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106 or any combination thereof, and a disclosed guide RNA sequence can comprise the sequence of (i) any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0245] In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise the sequence of SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, or a fragment thereof,or any combination thereof, and a disclosed guide RNA sequence can comprise the sequence of (i) any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0246] In an aspect of a disclosed 3’ replacement construct, a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106, or a fragment thereof, or any combination thereof, and a disclosed guide RNA sequence can comprise the sequence of (i) any one of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) any one of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) any one of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) any one of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0247] In an aspect of a disclosed 3’ replacement construct, one or more disclosed RNA structures can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO: 99 or any combination thereof, and one or more disclosed guide RNA sequences can comprise the sequence of (i) one or more of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) one or more of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) one or more of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) one or more of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0248] In an aspect of a disclosed 3’ replacement construct, one or more disclosed RNA structures can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106 or any combination thereof, and one or more disclosed guide RNA sequences can comprise the sequence of (i) one or more of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) one or more of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) one or more of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) one or more of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0249] In an aspect of a disclosed 3’ replacement construct, one or more disclosed RNA structures can comprise the sequence of any one of SEQ ID NO: 97 - SEQ ID NO: 99 or a fragment thereof, or any combination thereof, and one or more disclosed guide RNA sequences can comprise the sequence of (i) one or more of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) one or more of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) one or more of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) one or more of SEQ ID NO:30 - SEQ ID NO:43, or a fragment thereof, or any combination thereof.

[0250] In an aspect of a disclosed 3’ replacement construct, one or more disclosed RNA structures can comprise the sequence of any one of SEQ ID NO: 100 - SEQ ID NO: 106, or a fragment thereof,or any combination thereof, and one or more disclosed guide RNA sequences can comprise the sequence of (i) one or more of SEQ ID NO:01 - SEQ ID NO:29, or any combination thereof, (ii) one or more of SEQ ID NO:01 - SEQ ID NO:29, or fragment thereof, or any combination thereof, (iii) one or more of SEQ ID NO:30 - SEQ ID NO:43, or any combination thereof, or (iv) one or more of SEQ ID NO:30 - SEQ ID NO:43. or a fragment thereof, or any combination thereof.

[0251] In an aspect of a disclosed 3’ replacement construct, a disclosed combination of a disclosed RNA structure and a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:44 - SEQ ID NO:47.

[0252] In an aspect of a disclosed 3’ replacement construct, a disclosed combination can comprise a disclosed RNA structure followed by a disclosed guide RNA sequence. In an aspect of a disclosed 3’ replacement construct, a disclosed combination can comprise a disclosed guide RNA sequence followed by a disclosed RNA structure.

[0253] In an aspect of a disclosed 3’ replacement construct, a disclosed combination can comprise a disclosed RNA structure followed by a disclosed guide RNA sequence, wherein a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106, and wherein a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43. In an aspect of a disclosed 3’ replacement construct, a disclosed combination can comprise a disclosed RNA structure followed by a disclosed guide RNA sequence, wherein a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment thereof, and wherein a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 -SEQ ID NO:43 or a fragment thereof.

[0254] In an aspect of a disclosed 3’ replacement construct, a disclosed combination can comprise a disclosed guide RNA sequence followed by a disclosed RNA structure, wherein a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43, and wherein a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106. In an aspect of a disclosed 3’ replacement construct, a disclosed combination can comprise a disclosed guide RNA sequence followed by a disclosed RNA structure, wherein a disclosed guide RNA sequence can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 or a fragment thereof, and wherein a disclosed RNA structure can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment.

[0255] In an aspect of a disclosed 3’ replacement construct, the combination of two disclosed RNA structures and two disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:48 - SEQ ID NO:60. In an aspect of a disclosed 3‘ replacement construct, the combination of two disclosed RNA structures and two disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:48 - SEQ ID NO:60 or a fragment thereof.

[0256] In an aspect of a disclosed 3’ replacement construct, the combination of two disclosed RNA structures and two disclosed guide RNAs can comprise the sequence of any one of SEQ ID NO:48 - SEQ ID NO: 60, wherein the disclosed RNA structures can comprise the sequence of any one of SEQ ID NO: 97 - SEQ ID NO: 99 or SEQ ID NO: 100 - SEQ ID NO: 106, and the disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NQ:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43. In an aspect, a disclosed combination of two disclosed RNA structures and two disclosed RNA guides, the disclosed RNA structures can comprise the sequence of any one of SEQ ID NO: 97 - SEQ ID NO: 99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment, and the disclosed guide RNA sequences can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 or a fragment thereof.

[0257] In an aspect of a disclosed 3’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO:96. In an aspect of a disclosed 3' replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO:96 and be optimized by addition of one or more disclosed RNA structures and one or more disclosed guide RNAs. In an aspect of a disclosed 3’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO:96 and can be combined with one or more disclosed RNA structures and one or more disclosed guide RNAs. In an aspect of a disclosed 3’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO: 96 and can be combined with one or more disclosed RNA structures and one or more disclosed guide RNAs, wherein the disclosed RNA structures can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment, and wherein the disclosed guide RNAs can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 or a fragment thereof.

[0258] In an aspect of a disclosed 3’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise a sequence having at least 80%, at least 85%, at least 90%. at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:96. In an aspect of a disclosed 3’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:96 and can be combined with one ormore disclosed RNA structures and one or more disclosed guide RNAs. In an aspect of a disclosed 3’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:96 and can be combined with one or more disclosed RNA structures and one or more disclosed guide RNAs. wherein the disclosed RNA structures can comprise the sequence of any one of SEQ ID NO:97 - SEQ ID NO:99 or SEQ ID NO: 100 - SEQ ID NO: 106 or a fragment, and wherein the disclosed guide RNAs can comprise the sequence of any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43 or a fragment thereof.

[0259] In an aspect of a disclosed 3’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise the sequence set forth in SEQ ID NO: 86. In an aspect of a disclosed 3’ replacement construct, a disclosed exogenous RNA to be trans-spliced can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:86. In an aspect, Table 7 provides a listing of exemplary 3’ replacement constructs.Table 7 - List of Exemplary 3’ Replacement Constructs

[0260] In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more mutations or aberrations (such as, for example, those listed in Table 6). In an aspect of a disclosed targeted endogenous MYH7 pre-mRNA, one or more disclosed mutations or aberrations can be in the 5’ portion of the pre-mRNA. In an aspect, one or more disclosed mutations or aberrations in one or more exons can contribute to pathogenesis of one or more cells. In an aspect, one or more disclosed mutations or aberrations can inhibit translation of the encoded protein. In an aspect, one or more disclosed mutations or aberrations can modify translation of the encoded protein. In an aspect, one or more disclosed mutations or aberrations can generate an encoded protein having a non-sense mutation or a missense mutation.

[0261] In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more mutations or aberrations in one or more exons. In an aspect, a disclosed targeted endogenous MYH7 pre-mRNA can comprise one or more mutations or aberrations in one or more introns. In an aspect, one or more disclosed exonic mutations or aberrations can contribute to pathogenesisin one or more cells. In an aspect, one or more disclosed intronic mutations or aberrations can contribute to pathogenesis in one or more cells. In an aspect, a disclosed resulting chimeric RNA transcript can comprise the 5’ portion of the targeted endogenous pre-mRNA and the 3‘ portion of the exogenous RNA. In an aspect, a disclosed targeted endogenous pre-mRNA and a disclosed exogenous RNA can encode the same protein coding gene (i.e., MYH7). In an aspect, a disclosed targeted endogenous pre-mRNA and a disclosed exogenous RNA can comprise one or more exons of the same protein coding gene (i.e., MYH7). In an aspect, a disclosed 3’ replacement construct can redress one or more of the 165 mutations or aberrations identified below in Table 6.

[0262] In an aspect, a disclosed 3' hemi intron can comprise the sequence set forth in SEQ ID NO:90. In an aspect, a disclosed 3’ hemi intron can comprise a sequence having at least 80%, at least 85%, at least 90%, at least 95% identity, or more than 95% identity to the sequence set forth in SEQ ID NO:90. In an aspect, a disclosed 3’ hemi intron can be linked to a disclosed exogenous RNA to be trans-spliced. In an aspect, a disclosed 3’ hemi intron can comprise a branch point sequence that can recruit a U2 snRNP. In an aspect, a disclosed U2 snRNP can enable formation of a spliceosome with a disclosed exogenous RNA to be trans-spliced and a disclosed targeted endogenous MYH7 pre-mRNA. In an aspect, a disclosed 3’ hemi intron can be recognized by the spliceosome in a host cell. In an aspect, a disclosed 3’ hemi intron can be recognized by nuclear splicing components in a host cell.

[0263] In an aspect, proper assembly of a spliceosome between a disclosed exogenous RNA to be trans-spliced and a disclosed targeted endogenous MYH7 pre-mRNA can facilitate the joining of the 3’ end of the disclosed targeted endogenous MYH7 pre-mRNA with a disclosed exogenous RNA to be trans-spliced. In an aspect, proper assembly of a spliceosome between a disclosed exogenous RNA to be trans-spliced and a disclosed targeted endogenous MYH7 pre-mRNA can facilitate the joining of exons 1-22 of the disclosed targeted endogenous MYH7 pre-mRNA with exons 23-40 of the disclosed exogenous RNA to be trans-spliced. In an aspect, proper assembly of a spliceosome between a disclosed exogenous RNA to be trans-spliced and a disclosed targeted endogenous MYH7 pre-mRNA can facilitate the joining of exons 1-22 of the disclosed targeted endogenous MYH7 pre-mRNA with exons 23-40 of the disclosed exogenous RNA to be trans-spliced, thereby correcting and / or replacing one or more pathogenic mutations or aberrations in exons 23-40 of the targeted endogenous MYH7 pre-mRNA.

[0264] In an aspect, a disclosed 3’ hemi intron can facilitate the trans-splicing of the exogenous RNA to the exon immediately 3 ’ to the targeted intron in the endogenous pre-mRNA. In an aspect, a disclosed 3’ replacement construct can enable the 3’ replacement of the targeted endogenous pre-MYH7 mRNA having one or more mutations or aberrations. In an aspect, a disclosed 3’replacement construct can enable the replacement of exons 23-40 of the targeted endogenous MYH7 pre-mRNA.

[0265] In an aspect, a disclosed 3’ replacement construct can be used in atrans-splicing event. In an aspect, a disclosed 3’ replacement construct can be used in conjunction with one or more blocking U7 snRNAs. In an aspect, a disclosed 3’ replacement construct can further comprise the sequence of one or more blocking U7 snRNAs. In an aspect, a disclosed blocking U7 snRNA can be used to inhibit or decrease cis-splicing during a trans-splicing event comprising a disclosed 3’ replacement construct. In aspect, when used with one or more disclosed blocking U7 snRNAs, a disclosed 5‘ replacement construct can more efficiently drive trans-splicing. In an aspect, a disclosed blocking U7 snRNA can comprise the sequence set forth in any one of SEQ ID NO:62 - SEQ ID NO:71. In an aspect, a disclosed blocking U7 snRNA can comprise a sequence having at least 90% identity to the sequence set forth in any one of SEQ ID NO: 62 - SEQ ID NO:71. In an aspect, a disclosed U7 snRNA of SEQ ID NO:62 can be used with a disclosed guide RNA sequence of SEQ ID NO:01. In an aspect, a disclosed U7 snRNA of SEQ ID NO:63 can be used with a disclosed guide RNA sequence of SEQ ID NO:02. In an aspect, a disclosed U7 snRNA of SEQ ID NO: 64 can be used with a disclosed guide RNA sequence of SEQ ID NO: 03. In an aspect, a disclosed U7 snRNA of SEQ ID NO:65 can be used with a disclosed guide RNA sequence of SEQ ID NO:04. In an aspect, a disclosed U7 snRNA of SEQ ID NO:66 can be used with a disclosed guide RNA sequence of SEQ ID NO:05. In an aspect, a disclosed U7 snRNA of SEQ ID NO: 67 can be used with a disclosed guide RNA sequence of SEQ ID NO: 06. In an aspect, a disclosed U7 snRNA of SEQ ID NO:68 can be used with a disclosed guide RNA sequence of SEQ ID NO:07. In an aspect, a disclosed U7 snRNA of SEQ ID NO:69 can be used with a disclosed guide RNA sequence of SEQ ID NO:08. In an aspect, a disclosed U7 snRNA of SEQ ID NO:70 can be used with a disclosed guide RNA sequence of SEQ ID NO:09. In an aspect, a disclosed U7 snRNA of SEQ ID NO: 71 can be used with a disclosed guide RNA sequence of SEQ ID NO: 10.

[0266] In an aspect, a disclosed U7 snRNA expression construct comprising a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator can comprise the sequence set forth in any one of SEQ ID NO: 73 - SEQ ID NO: 82. In an aspect, a disclosed U7 snRNA expression construct comprising a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator can comprise a sequence having at least 90% to the sequence set forth in any one of SEQ ID NO: 73 - SEQ ID NO: 82.

[0267] In an aspect, a disclosed U7 snRNA expression construct comprising a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator can comprise thesequence set forth in any one of SEQ ID NO:73 - SEQ ID NO:82, and can be used in conjunction with one or more 3’ replacement constructs.

[0268] In an aspect, a disclosed U7 snRNA expression construct comprising a disclosed U7 snRNA, a U7 promoter, a U7 smOPT stem loop, and a U7 RNA terminator can comprise a sequence having at least 90% to the sequence set forth in any one of SEQ ID NO: 73 - SEQ ID NO:82, and can be used in conjunction with one or more 3’ replacement constructs.

[0269] In an aspect, a disclosed U7 snRNA expression construct can be incorporated into a viral vector or a non-viral vector. In an aspect, a disclosed U7 snRNA expression construct can be incorporated into a viral vector or a non-viral vector comprising a disclosed nucleic acid molecule.

[0270] In an aspect, a disclosed promoter for the 3’ replacement construct can be tissue-specific or ubiquitous and can be constitutive or inducible, depending on the pattern of the expression desired. In an aspect, a disclosed promoter can be native or foreign and can be a natural or a synthetic sequence. By foreign, it is intended that the transcriptional initiation region is not found in the wild-type host into which the transcriptional initiation region is introduced. In an aspect, a disclosed promoter can be a promoter / enhancer. In an aspect, a disclosed promoter for the disclosed nucleic acid molecule can be an endogenous promoter. In an aspect, a disclosed endogenous promoter can be an endogenous promoter / enhancer. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can generally be obtained from a non-coding region upstream of a transcription initiation site of a gene of interest. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can be used for constitutive and efficient expression of a disclosed gene. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be a CMV promoter or a CMV promoter / enhancer. In an aspect, a disclosed CMV protomer can compnse the sequence set forth in SEQ ID NO:83. CMV promoters and CMV promoters / enhancers are well known to the art. In an aspect, a disclosed promoter for the one or more disclosed guide RNA sequences can be any eukaryotic RNA polymerase II promoter.

[0271] In an aspect, a disclosed nucleic acid molecule can further comprise can further comprise a UTR. In an aspect, a disclosed nucleic acid molecule can further comprise a polyadenylation sequence. In an aspect, a disclosed nucleic acid molecule can further comprise a sequence for a promoter. In an aspect, a disclosed nucleic acid molecule can further comprise a spacer region. In an aspect, a disclosed spacer region can separate the 5' splice region from the one or more RNA structures. In an aspect, a disclosed spacer region can comprise any known spacer. In an aspect, a disclosed spacer region can comprise a consensus splicing motif (e.g., such as U1 or U2). In an aspect, a disclosed spacer region can comprise a limited number of consensus splicing motifs (e.g., such as U1 or U2).

[0272] In an aspect, a disclosed nucleic acid molecule can further comprise one or more nuclear localization signals (NLS). NLS are known to the skilled person in the art. In an aspect, a disclosed NLS can comprise any NLS known to the art. As known to the art (see, e.g., Lu J, et al. (2021) Cell Commun Signal. 19:60, which is incorporated herein by reference for its teachings of NLS), nuclear localization signals (NLS) are generally short peptides that act as a signal fragment that mediates the transport of proteins from the cytoplasm into the nucleus.

[0273] In an aspect, a disclosed nucleic acid molecule can further comprise one or more nuclear retention elements (NRE). NRE are known to the skilled person in the art. In an aspect, a disclosed NRE can comprise SIRLOIN (SEQ ID NO: 113) or BORG (SEQ ID NO: 114).

[0274] In an aspect, a disclosed nucleic acid molecule can further comprise one or more Flavivirus genetic elements. In an aspect, Flavivirus genetic elements can comprise one or more Flavivirus 3’ untranslated region (3' UTR), one or more subgenomic Flavivirus RNA (siRNA) elements, one or more Flavivirus XRN1 -resistant RNA (xrRNA) elements, one or more Flavivirus dumbbell (DB) RNA elements, one or more Flavivirus 3’ stem loop (3’ SL) elements, or any combination thereof. (See WO 2022 / 182835 for a description of Flavivirus gene elements).

[0275] In an aspect, a disclosed cell or a disclosed host cell can be in a subject. In an aspect, a subject can be a human patient and can be male or female. In an aspect, a subject can have a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations). In an aspect, a subject can be treatment-naive. In an aspect, a patient can refer to a subject afflicted with a disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations). In an aspect, a subject or a patient can be seeking treatment or receiving treatment for a disease or disorder (i.e., hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations).

[0276] In an aspect, a disclosed nucleic acid molecule comprising a 3’ replacement construct can be packaged into lipid nanoparticles. In an aspect, a disclosed nucleic acid molecule comprising a 3’ replacement construct can be packaged into a viral vector. In an aspect, a disclosed viral vector can comprise an AAV vector. In an aspect, a disclosed nucleic acid molecule can be packaged into a non-viral carrier. In an aspect, a disclosed nucleic acid molecule can be incorporated into a plasmid. In an aspect, a disclosed nucleic acid molecule can be incorporated into lipid nanoparticles.

[0277] Disclosed herein is an expression cassette comprising disclosed 3 ’ replacement construct. Disclosed herein is an expression cassette comprising a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3 ’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA. Disclosed herein is an expression cassette comprising a nucleic acid molecule comprisinga promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA.

[0278] Disclosed herein is an expression cassette comprising (1) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; or (2) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA.

[0279] Disclosed herein is an expression cassette comprising (1) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (2) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (3) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA; (4) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (5) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; a guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; (6) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; (7) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one ormore guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (8) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3‘ portion of the targeted endogenous MYH7 pre-mRNA; (9) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (10) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (11) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA; (12) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (13) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (14) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (15) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (16) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (17) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (18) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (19) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3 ’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (20) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be transspliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (21) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; or (22) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0280] In an aspect, use of a disclosed 3’ replacement construct or a disclosed expression cassette comprising a disclosed 3’ replacement construct can be used to restore and / or return MYH7 expression to a wild-type, normal, or control expression level. In an aspect, a disclosed nucleic acid molecule can restore one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation. In an aspect, a disclosed nucleic acid molecule can restore the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme (i.e., MYH7). In an aspect, restoring one or more aspects of cellular homeostasis and / or cellular functionality can comprise one or more of the following: (i) correcting cell starvation in one or more cell ty pes; (ii) normalizing aspects of the autophagy7pathway (such as, for example, correcting, preventing, reducing, and / or ameliorating autophagy); (iii) improving, enhancing, restoring, and / or preserving mitochondrial functionality and / or structural integrity; (iv) improving, enhancing, restoring, and / or presenring organelle functionality and / or structural integrity7; (v) correcting enzyme dysregulation; (vi) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of the multi-systemic manifestations of a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations); (vii) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations); or (viii) any combination thereof. In an aspect, restoringone or more aspects of cellular homeostasis can comprise improving, enhancing, restoring, and / or preserving one or more aspects of cellular structural and / or functional integrity.

[0281] In an aspect, restoring the activity and / or functionality of a missing, deficient, and / or mutant protein or enzyme (i.e., MYH7) can comprise a 10%, 20%. 30%. 40%, 50%, 60%, 70%, 80%. 90%. 100%, or any amount of restoration when compared to a pre-existing level such as, for example, a pre-treatment level. In an aspect, the amount of restoration can be 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% more than a pre-existing level such as, for example, a pre-treatment level. In an aspect, restoration can be measured against a control level or a reference level (e.g., determined, for example, using one or more subjects not having a missing, deficient, and / or mutant protein or enzyme like MYH7). In an aspect, restoration can be a partial or incomplete restoration. In an aspect, restoration can be complete or near complete restoration such that the level of expression, activity, and / or functionality is similar to that of a wild-type or control level.

[0282] In an aspect, a disclosed nucleic acid sequence can further comprising the sequence of one or more disclosed blocking U7 snRNAs. In an aspect, a disclosed 3’ replacement construct can further comprise the sequence of one or more disclosed blocking U7 snRNAs. In an aspect, a disclosed nucleic acid sequence comprising a 3’ replacement construct can further comprise the sequence of one or more disclosed blocking U7 snRNAs.4. Vectors

[0283] Disclosed herein is a vector comprising a disclosed nucleic acid molecule. Disclosed herein is a non-viral vector comprising a disclosed nucleic acid molecule. Disclosed herein is a viral vector comprising a disclosed nucleic acid molecule. Disclosed herein is a non-viral vector comprising one or more disclosed nucleic acid molecules. Disclosed herein is a viral vector comprising one or more disclosed nucleic acid molecules. Disclosed herein is a non-viral or viral vector comprising a disclosed 5’ replacement construct. Disclosed herein is anon-viral or viral vector comprising a disclosed 3' replacement construct. Disclosed herein is a non-viral or viral vector comprising one or more disclosed 5’ replacement constructs. Disclosed herein is a non-viral or viral vector comprising one or more disclosed 3’ replacement constructs.

[0284] Disclosed herein is a viral vector or non-viral vector comprising a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a viral vector or non-viral vector comprising a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5‘ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.Disclosed herein is a viral vector or non-viral vector comprising (1) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (2) a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (3) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (4) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (5) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (6) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (7) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (8) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (9) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (10) a multivalent guide trans-splicing RNA molecule comprising a promoter; anexogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5' portion of the targeted endogenous MYH7 pre-mRNA; (11) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (12) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (13) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (14) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (15) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (16) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (17) a nucleic acid molecule anexogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (18) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (19) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (20) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (21) a multivalent guide trans-splicing RNA molecule comprising a promoter an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (22) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-splicedto a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; or (23) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures: and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA.

[0285] Disclosed herein is a viral vector or non-viral vector comprising (1) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; or (2) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA.

[0286] Disclosed herein is a viral vector or non-viral vector comprising (1) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (2) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (3) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA; (4) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targetedendogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (5) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; a guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; (6) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; (7) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (8) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3‘ portion of the targeted endogenous MYH7 pre-mRNA; (9) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (10) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (11) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous pre-mRNA; (12) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targetedendogenous MYH7 pre-mRNA; (13) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3 ’ portion of the targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (14) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3‘ hemi intron linked to an exogenous RNA to be trans-spliced: and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (15) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (16) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (17) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (18) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (19) a multivalent guide trans-splicing RNA molecule comprising one or more RNAstructures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3 ’ portion of the targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (20) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (21) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; or (22) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA.

[0287] Disclosed herein is a vector comprising a nucleic acid sequence comprising the sequence of one or more blocking U7 snRNAs. Disclosed herein is an AAV vector comprising a nucleic acid sequence comprising the sequence of one or more blocking U7 snRNAs.

[0288] In an aspect, a disclosed vector can be formulated for administration via one or more routes. Such methods are well known to those skilled in the art and include, but are not limited to, the following routes: oral administration, transdermal administration, administration by inhalation, nasal administration, topical administration, in utero administration, intrahepatic administration, intravaginal administration, ophthalmic administration, intraaural administration, otic administration, intracerebral administration, rectal administration, sublingual administration, buccal administration, and parenteral administration, including injectable such as intravenousadministration, intra-CSF administration, intra-arterial administration, intramuscular administration, and subcutaneous administration. Administration can also include hepatic intraarterial administration or administration through the hepatic portal vein (HPV). Administration of a disclosed therapeutic agent, a disclosed pharmaceutical composition, or a combination thereof can comprise administration directly into the CNS (e.g., intraparenchymal, intracerebroventricular, intrathecal cisternal, intrathecal (lumbar), deep gray matter delivery, convection-enhanced delivery to deep gray matter) or the PNS. Administration can be continuous or intermittent. Administration can be intracardiac. Administration can comprise administering a viral vector and / or generated optimized viral vector. Administration of a disclosed vector can be continuous or intermittent.

[0289] In an aspect, a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV vector can comprise a range of about 1 x IO10vg / kg to about 2 x 1014.vg / kg. In an aspect, for example, a disclosed vector can be administered at a dose of about 1 x 1011to about 8 x 1013vg / kg or about 1 x 1012to about 8 x 1013vg / kg. In an aspect, a disclosed vector can be administered at a dose of about 1 x 1013to about 6 x 1013vg / kg. In an aspect, a disclosed vector can be administered at a dose of at least about 1 x IO10, at least about 5 x IO10, at least about 1 x 1011, at least about 5 x 1011, at least about 1 x IO12, at least about 5 x 1012, at least about 1 x 1013, at least about 5 x 1013, or at least about 1 x 1014vg / kg. In an aspect, a disclosed vector can be administered at a dose of no more than about 1 x IO10, no more than about 5 x IO10, no more than about 1 x 1011, no more than about 5 x 1011, no more than about 1 x 1012, no more than about 5 x 1012, no more than about 1 x 1013, no more than about 5 x 1013, or no more than about 1 x 1014vg / kg. In an aspect, a disclosed vector can be administered at a dose of about 1 x 1012vg / kg. In an aspect, a disclosed vector can be administered at a dose of about 1 x 1011vg / kg. In an aspect, a disclosed vector can be administered in a single dose, or in multiple doses (such as 2, 3, 4, 5, 6, 7, 8, 9 or 10 doses) as needed for the desired therapeutic results.

[0290] In an aspect, a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV vector can comprise a range of about I x 1012vg per subject total to about 1 x 1017vg per subject total. In an aspect, a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV vector can comprise a range of about 1 x 1012vgper subject total, about 1 x 1013vg per subject total, about 1 x 1014vg per subject total, about 1 x 1015vg per subject total, about 1 x 1016vg per subject total, or about 1 x 1017vg per subject total. In an aspect, a therapeutically effective amount of a disclosed AAV particle or a disclosed AAV vector can be delivered retrograde ureteral infusion and / or renal arterial administration and can comprise a range of about 1 x 1012vg per subject total to about 1 x 1017vg per subject total.

[0291] In an aspect, a therapeutically effective amount of a disclosed AAV particle can comprise about 1 x 106DRP / mL to about 1 x 1014DRP / mL. In an aspect, a disclosed pharmaceutical formulation can comprise about 1 x 106DRP / mL, 1 x 107DRP / mL, 1 x 108DRP / mL, 1 x 109DRP / mL, 1 x IO10DRP / mL, 1 x 1011DRP / mL, 1 x 1012DRP / mL, 1 x 1013DRP / mL, or 1 x 1014DRP / mL. In an aspect, a therapeutically effective amount of a disclosed AAV particle or a disclosed vector can comprise a range determined by a skilled person.

[0292] In an aspect, a disclosed non-viral vector can be a polymer-based vector, a peptide-based vector, a lipid nanoparticle, a solid lipid nanoparticle, or a cationic lipid-based vector. In an aspect, a disclosed vector can comprise exosomes, extracellular vesicles, and virus like particles.

[0293] In an aspect, a disclosed viral vector can be an adenovirus vector, an AAV vector, a herpes simplex virus vector, a retrovirus vector, a lentivirus vector, and alphavirus vector, a Flavivirus vector, a rhabdovirus vector, a measles virus vector, a Newcastle disease viral vector, a poxvirus vector, or a picomavirus vector. In an aspect, a disclosed nucleic acid sequence can have a coding sequence that is less than about 4.5 kilobases.

[0294] In an aspect, a disclosed AAV vector can include naturally isolated serotypes including, but not limited to, AAV1, AAV2, AAV3 (including 3a and 3b), AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh8, AAV9, AAV10, AAVrhlO. AAV11, AAV12, AAV13, AAVrh39, AAVrh43, AAVcy.7 as well as bovine AAV. caprine AAV, canine AAV. equine AAV, ovine AAV. avian AAV, primate AAV, non-primate AAV, and any other virus classified by the International Committee on Taxonomy of Viruses (ICTV) as an AAV. In an aspect, an AAV capsid can be a chimera either created by capsid evolution or by rational capsid engineering from a naturally isolated AAV variants to capture desirable serotype features such as enhanced or specific tissue tropism and / or a host immune response escape. Naturally isolated AAV variants include, but not limited to, AAV-DJ, AAV-HAE1, AAV-HAE2, AAVM41, AAV- 1829, AAV2 Y / F, AAV2 T / V, AAV2i8, AAV2.5, AAV9.45, AAV9.61, AAV-B1, AAV-AS, AAV9.45A-String (e.g., AAV9.45-AS). AAV9.45Angiopep, AAV9.47-Angiopep, and AAV9.47-AS, AAV-PHP.B, AAV-PHP.eB. AAV-PHP.S, AAV-F, AAVcc.47. and AAVcc.81. In an aspect, a disclosed AAV vector can be AAV-Rh74 or a related variant (e.g., capsid variants like RHM4-1). In an aspect, a disclosed AAV vector can be AAV.cc47. In an aspect, adisclosed AAV vector can be AAV.cc81. In an aspect, a disclosed AAV vector can be a self-complementary' AAV.

[0295] In an aspect, a disclosed vector can comprise one or more ITRs (such as, for example, ITRs from AAV2).

[0296] In an aspect, a disclosed vector can further comprise one or more nuclear localization signals (NLS). NLS are known to the skilled person in the art. In an aspect, a disclosed NLS can comprise any NLS known to the art. As known to the art (see, e.g., Lu J, et al. (2021) CellCommun Signal. 19:60, which is incorporated herein by reference for its teachings of NLS), nuclear localization signals (NLS) are generally short peptides that act as a signal fragment that mediates the transport of proteins from the cytoplasm into the nucleus.

[0297] In an aspect, a disclosed vector can further comprise one or more nuclear retention elements (NRE). NRE are known to the skilled person in the art. In an aspect, a disclosed NRE can comprise SIRLOIN (SEQ ID NO: 113) or BORG (SEQ ID NO: 114).

[0298] In an aspect, a disclosed vector can further comprise one or more Flavivirus genetic elements. In an aspect, Flavivirus genetic elements can comprise one or more Flavivirus 3’ untranslated region (3’ UTR), one or more subgenomic Flavivirus RNA (sfRNA) elements, one or more Flavivirus XRN1 -resistant RNA (xrRNA) elements, one or more Flavivirus dumbbell (DB) RNA elements, one or more Flavivirus 3’ stem loop (3’ SL) elements, or any combination thereof. (See WO 2022 / 182835 for a description of Flavivirus gene elements).

[0299] In an aspect, a disclosed vector can further comprise a nucleic acid sequence encoding a therapeutic protein, a therapeutic agent, and / or a therapeutic RNA. In an aspect, a disclosed therapeutic protein can comprise a polypeptide and / or a glycopeptide. In an aspect, a disclosed therapeutic agent can comprise an oligonucleotide therapeutic agent. In an aspect, a disclosed oligonucleotide therapeutic agent can be a single-stranded or double-stranded DNA, iRNA, shRNA, siRNA, mRNA. non-coding RNA (ncRNA), an antisense molecule, miRNA, a morpholino, a peptide-nucleic acid (PNA), or an analog or conjugate thereof. In an aspect, a disclosed therapeutic agent can be an ASO or an RNAi. In an aspect, a disclosed therapeutic agent can comprise a CRISPR-based endonuclease (e.g., Cas9). In an aspect, a disclosed CRISPR-based endonuclease can be derived from a CRISPR / Cas type I, type II, or type III system.

[0300] In an aspect, a disclosed therapeutic RNA can comprise ribosomal RNA (rRNA), transfer RNA (tRNA), heterogeneous nuclear RNA (hnRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), micro RNA (miRNA), Piwi-interacting RNA (piRNA), small interfering RNA (siRNA), short hairpin RNA (shRNA), singe guide RNA (sgRNA), non-coding RNA (ncRNA), long non-coding RNA (IncRNA), 7SL, Xist, short enhancer RNA (eRNA), circular RNA, intergenic RNA, or any combination thereof. In an aspect, a disclosed RNA can comprise IncRNA, siRNA, shRNA, sgRNA, circular RNA, snoRNA, miRNA, or any combination thereof. In an aspect, a disclosed encoded RNA can comprise a functional non-coding RNA element.

[0301] In an aspect, a disclosed vector can comprise one or more promoters operably linked to a disclosed nucleic acid molecule (e.g., a 5’ replacement construct and / or 3’ replacement construct), a disclosed transgene, a disclosed sequence to be trans-spliced, and / or a disclosed nucleic acid sequence. In an aspect of a disclosed vector, a disclosed nucleic acid molecule can be operablylinked to one or more transcription regulatory elements. In an aspect, the one or more transcription regulatory elements (e.g., Woodchuck Hepatitis Virus (WHV) Posttranscriptional Regulator Element (WPRE), triplex from MALAT1, the PRE of Hepatitis B virus (HPRE), and an iron response element) can increase the transcription and / or expression of a disclosed nucleic acid molecule (e.g.. a 5’ replacement construct and / or 3’ replacement construct), a disclosed transgene, a disclosed sequence to be trans-spliced, and / or a disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA).

[0302] In an aspect, a disclosed promoter can be positioned 5’ (upstream) or 3’ (downstream) of a disclosed nucleic acid molecule (e.g., a 5’ replacement construct and / or 3' replacement construct), a disclosed transgene, a disclosed sequence to be trans-spliced, and / or a disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA) under its control. The distance between a disclosed promoter and a disclosed nucleic acid molecule (e.g., a 5' replacement construct and / or 3’ replacement construct), a disclosed transgene, a disclosed sequence to be trans-spliced, and / or a disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA) can be approximately the same as the distance between that promoter and to the disclosed nucleic acid molecule (e.g., a 5’ replacement construct and / or 3‘ replacement construct), the disclosed transgene, the disclosed sequence to be trans-spliced, and / or the disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA) under its control. As is known in the art, variation in this distance can be accommodated w ithout loss of promoter function.

[0303] In an aspect of a disclosed vector, a disclosed promoter can be tissue-specific or ubiquitous and can be constitutive or inducible, depending on the pattern of the expression desired. A disclosed promoter can be native or foreign and can be a natural or a synthetic sequence. By foreign, it is intended that the transcriptional initiation region is not found in the wild-type host into which the transcriptional initiation region is introduced. In an aspect, a disclosed promoter can be a promoter / enhancer. In an aspect, a disclosed promoter for a disclosed nucleic acid molecule (e.g., a 5’ replacement construct and / or 3’ replacement construct), a disclosed transgene, a disclosed sequence to be trans-spliced, and / or a disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA) can be an endogenous promoter. In an aspect, a disclosed endogenous promoter can be an endogenous promoter / enhancer. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can generally be obtained from a non-coding region upstream of a transcription initiation site of a gene of interest. In an aspect, a disclosed endogenous promoter or a disclosed endogenous promoter / enhancer can be used for constitutive and efficient expression of a disclosed protein coding gene. In an aspect, a disclosed promoter for a disclosed nucleic acid molecule(e.g., a 5’ replacement construct and / or 3’ replacement construct), a disclosed transgene, a disclosed sequence to be trans-spliced, and / or a disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA) can be a CMV promoter or a CMV promoter / enhancer. CMV promoters and CMV promoters / enhancers are well known to the art. In an aspect, a disclosed promoter for a disclosed nucleic acid molecule (e.g., a 5’ replacement construct and / or 3’ replacement construct), a disclosed transgene, a disclosed sequence to be transspliced, and / or a disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA) can be any eukaryotic RNA polymerase II promoter.

[0304] In an aspect, a disclosed AAV vector can be used to generate AAV particles. In an aspect, a disclosed AAV vector can be used to generate AAV particles comprising a disclosed nucleic acid molecule (e.g., a 5’ replacement construct and / or 3’ replacement construct), a disclosed transgene, a disclosed sequence to be trans-spliced, and / or a disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA) under its control.

[0305] Disclosed herein is an AAV particle comprising a disclosed nucleic acid molecule (e.g., a 5’ replacement construct and / or 3’ replacement construct), a disclosed transgene, a disclosed sequence to be trans-spliced, and / or a disclosed nucleic acid sequence (e.g., encoding a disclosed therapeutic protein and / or a disclosed therapeutic RNA) under its control.

[0306] In an aspect, a disclosed vector or a disclosed AAV particle can be particularly useful in a method of treating a subject having one or more mutations or aberrations in the MYH7 gene.

[0307] Disclosed herein is a vector comprising a disclosed nucleic acid sequence comprising the sequence of one or more disclosed blocking U7 snRNAs. Disclosed herein is a vector comprising a disclosed 5’ replacement construct that further comprises the sequence of one or more disclosed blocking U7 snRNAs. Disclosed herein is a vector comprising a disclosed nucleic acid sequence comprising a 5’ replacement construct and further comprising the sequence of one or more disclosed blocking U7 snRNAs.

[0308] Disclosed herein is a vector comprising a disclosed nucleic acid sequence comprising the sequence of one or more disclosed blocking U7 snRNAs. Disclosed herein is a vector comprising a disclosed 3’ replacement construct that further comprises the sequence of one or more disclosed blocking U7 snRNAs. Disclosed herein is a vector comprising a disclosed nucleic acid sequence comprising a 3’ replacement construct and further comprising the sequence of one or more disclosed blocking U7 snRNAs.5. Pharmaceutical Formulations

[0309] Disclosed herein is a pharmaceutical formulation comprising a disclosed nucleic acid molecule. Disclosed herein is a pharmaceutical formulation comprising a disclosed nucleic acid molecule and one or more pharmaceutically acceptable carriers, diluents, and / or excipients.Disclosed herein is a pharmaceutical formulation comprising a disclosed vector. Disclosed herein is a pharmaceutical formulation comprising a disclosed vector and one or more pharmaceutically acceptable carriers, diluents, and / or excipients. Disclosed herein is a pharmaceutical formulation comprising a disclosed AAV particle. Disclosed herein is a pharmaceutical formulation comprising a disclosed AAV particle and one or more pharmaceutically acceptable carriers, diluents, and / or excipients. Disclosed herein is a pharmaceutical formulation comprising a non-viral vector or a viral vector comprising a disclosed nucleic acid molecule and one or more pharmaceutically acceptable carriers, diluents, and / or excipients.

[0310] Disclosed herein is a pharmaceutical formulation comprising a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a pharmaceutical formulation comprising a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a pharmaceutical formulation comprising a viral vector or non-viral vector comprising a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences. Disclosed herein is a pharmaceutical formulation comprising a viral vector or non-viral vector comprising a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA; a 5' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences.

[0311] Disclosed herein is a pharmaceutical formulation comprising (1) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (2) a nucleic acid molecule comprising a promoter, an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (3) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous MYH7 pre-mRNA; (4) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (5) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (6) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (7) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (8) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences; (9) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (10) a multivalent guide trans-splicing RNA molecule composing a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA; (11) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (12) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be transspliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (13) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be transspliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (14) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (15) a nucleic acid molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (16) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (17) a nucleic acid molecule an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (18) a nucleic acid molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ' hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (19) a multivalent guide trans-splicing RNA moleculecomprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (20) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5 ’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations or aberrations in the targeted endogenous MYH7 pre-mRNA; (21) a multivalent guide trans-splicing RNA molecule comprising a promoter an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 5’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (22) a multivalent guide trans-splicing RNA molecule comprising an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; or (23) a multivalent guide trans-splicing RNA molecule comprising a promoter; an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5’ hemi intron linked to an exogenous RNA to be trans-spliced; one or more RNA structures; and one or more guide RNA sequences, wherein the exogenous RNA to be trans-spliced comprises the first half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA.

[0312] Disclosed herein is a pharmaceutical formulation comprising (1) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; or (2) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA.

[0313] Disclosed herein is a pharmaceutical formulation comprising (1) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced: and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (2) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (3) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous pre-mRNA; (4) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (5) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; a guide RNA sequences: a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; (6) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; (7) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion ofthe targeted endogenous MYH7 pre-mRNA; (8) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be transspliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be transspliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA; (9) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (10) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA; (11) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous pre-mRNA; (12) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (13) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (14) a nucleic acid molecule comprising a promoter, one or more RNA structures: one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA;(15) a nucleic acid molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA. wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (16) a nucleic acid molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA;(17) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (18) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (19) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (20) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3' hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises one or more exons located in the 3’ portion of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; (21) a multivalent guide trans-splicing RNA molecule comprising one or more RNA structures; one ormore guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA; or (22) a multivalent guide trans-splicing RNA molecule comprising a promoter, one or more RNA structures; one or more guide RNA sequences; a 3’ hemi intron linked to an exogenous RNA to be trans-spliced; and an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced comprises the second half of the open reading frame of the targeted endogenous MYH7 pre-mRNA, wherein the exogenous RNA to be trans-spliced is designed to correct multiple pathogenic mutations in the targeted endogenous MYH7 pre-mRNA.

[0314] In an aspect, a disclosed pharmaceutical formulation can comprise (i) one or more active agents, (ii) biologically active agents, (iii) one or more pharmaceutically active agents, (iv) one or more immune-based therapeutic agents, (v) one or more clinically approved agents, or (vi) a combination thereof. In an aspect, a disclosed composition can comprise one or more immune modulators. In an aspect, a disclosed composition can comprise one or more proteasome inhibitors. In an aspect, a disclosed composition can comprise one or more immunosuppressives or immunosuppressive agents. In an aspect, an immunosuppressive agent can be anti-thymocyte globulin (ATG), cyclosporine (CSP), mycophenolate mofetil (MMF), or a combination thereof. In an aspect, a disclosed formulation can comprise an anaplerotic agent (such as, for example, C7 compounds like triheptanoin or MCT).

[0315] In an aspect, a disclosed formulation can comprise an RNA therapeutic. An RNA therapeutic can comprise RNA-mediated interference (RNAi) and / or antisense oligonucleotides (ASO). In an aspect, a disclosed RNA therapeutic can be directed at any protein or enz me that is overexpressed or is overactive due to a missing, deficient, and / or mutant protein or enzyme. In an aspect, a disclosed RNA therapeutic can comprise therapy delivered via LNPs. In an aspect, a disclosed formulation can comprise an enzyme or enzyme precursor for enzyme replacement therapy (ERT).

[0316] In an aspect, a disclosed formulation can comprise a disclosed small molecule. In an aspect, a disclosed small molecule can assist in restoring the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme.

[0317] In an aspect, any disclosed pharmaceutical formulation can comprise one or more excipients and / or pharmaceutically acceptable carriers. Excipients and / or pharmaceutically acceptable carriers are known to the art and are discussed supra.

[0318] In an aspect, a therapeutically effective amount of a disclosed pharmaceutical formulation can comprise a range of about 1 x IO10vg / kg to about 2 x 1014vg / kg of a disclosed vector and / or a disclosed AAV particle. In an aspect, for example, a dose of a disclosed pharmaceutical formulation can comprise about 1 x 1011to about 8 x 1013vg / kg or about 1 x 1012to about 8 x 1013vg / kg. In an aspect, a dose of a disclosed pharmaceutical formulation can comprise about 1 x 1013to about 6 x 1013vg / kg. In an aspect, a dose of a disclosed pharmaceutical formulation can comprise at least about 1 x IO10, at least about 5 x IO10, at least about 1 x 1011, at least about 5 x 1011, at least about 1 x 1012, at least about 5 x 1012, at least about 1 x 101?, at least about 5 x 101?, or at least about 1 x 1014vg / kg. In an aspect, a dose of a disclosed pharmaceutical formulation can comprise no more than about 1 x IO10, no more than about 5 x IO10, no more than about 1 x 1011, no more than about 5 x 1011, no more than about 1 x 1012, no more than about 5 x 1012, no more than about 1 x 1013, no more than about 5 x 1013, or no more than about 1 x 1014vg / kg. In an aspect, a dose of a disclosed pharmaceutical formulation can comprise about 1 x 1012vg / kg. In an aspect, a dose of a disclosed pharmaceutical formulation can comprise about 1 x 1011vg / kg. In an aspect, a dose of a disclosed pharmaceutical formulation can comprise a single dose, or in multiple doses (such as 2, 3, 4, 5, 6, 7, 8, 9 or 10 doses) as needed for the desired therapeutic results.

[0319] In an aspect, a therapeutically effective amount of a disclosed pharmaceutical formulation can comprise a range of about 1 x 1012vg per subject total to about 1 x 1017vg per subject total. In an aspect, a therapeutically effective amount of a disclosed pharmaceutical formulation can comprise a range of about 1 x 1012vg per subject total, about 1 x 1013vg per subject total, about 1 x 1014vg per subject total, about 1 x 1015vg per subject total, about 1 x 1016vg per subject total, or about 1 x 1017vg per subject total. In an aspect, a therapeutically effective amount of a disclosed pharmaceutical formulation can comprise about 1 x 106DRP / mL to about 1 x 1014DRP / mL. In an aspect, a therapeutically effective amount of a disclosed pharmaceutical formulation can comprise about 1 x 106DRP / mL, 1 x 107DRP / mL, 1 x 108DRP / mL, 1 x 109DRP / mL, 1 x IO10DRP / mL, 1 x 1011DRP / mL. 1 x 1012DRP / mL, 1 x 1013DRP / mL, or 1 x 1014DRP / mL.

[0320] In an aspect, a therapeutically effective amount of a disclosed pharmaceutical formulation can comprise a range determined by a skilled person.

[0321] In an aspect, a disclosed pharmaceutical formulation can be used to restore and / or return expression of a disclosed protein coding gene to a wild-type, normal, or control expression level. In an aspect, a disclosed pharmaceutical formulation can restore one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation. In an aspect, a disclosed nucleic acid molecule can restore the functionality and / or structural integrity of a missing, deficient, and / or mutant protein or enzyme (such as those, for example, encoded by one of thegenes provided supra). In an aspect, restoring one or more aspects of cellular homeostasis and / or cellular functionality can comprise one or more of the following: (i) correcting cell starvation in one or more cell types; (ii) normalizing aspects of the autophagy pathway (such as, for example, correcting, preventing, reducing, and / or ameliorating autophagy); (iii) improving, enhancing, restoring, and / or preserving mitochondrial functionality and / or structural integrity; (iv) improving, enhancing, restoring, and / or preserving organelle functionality and / or structural integrity; (v) correcting enzyme dysregulation; (vi) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of the multi-systemic manifestations of a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations); (vii) reversing, inhibiting, preventing, stabilizing, and / or slowing the rate of progression of a genetic disease or disorder (such as hypertrophic cardiomyopathy caused by one or more MYH7 mutations or aberrations); or (viii) any combination thereof. In an aspect, restoring one or more aspects of cellular homeostasis can comprise improving, enhancing, restoring, and / or preserving one or more aspects of cellular structural and / or functional integrity. In an aspect, restoring the activity and / or functionality of a missing, deficient, and / or mutant protein or enzyme can comprise a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or any amount of restoration when compared to a pre-existing level such as, for example, a pre-treatment level. In an aspect, the amount of restoration can be 10-20%, 20-30%, 30-40%, 40-50%, 50-60%, 60-70%, 70-80%, 80-90%, or 90-100% more than a pre-existing level such as, for example, a pre-treatment level. In an aspect, restoration can be measured against a control level or a reference level (e.g., determined, for example, using one or more subj ects not having a missing, deficient, and / or mutant protein or enzy me). In an aspect, restoration can be a partial or incomplete restoration. In an aspect, restoration can be complete or near complete restoration such that the level of expression, activity7, and / or functionality is similar to that of a wild-type or control level.

[0322] In an aspect, a disclosed pharmaceutical formulation can be particularly useful in a method of treating a subj ect having one or more mutations or variations in the MYH7 gene.

[0323] Disclosed herein is a pharmaceutical formulation comprising a vector comprising a disclosed nucleic acid sequence comprising the sequence of one or more disclosed blocking U7 snRNAs. Disclosed herein is a pharmaceutical formulation comprising a vector comprising a disclosed 5’ replacement construct that further comprises the sequence of one or more disclosed blocking U7 snRNAs. Disclosed herein is a pharmaceutical formulation comprising a vector comprising a disclosed nucleic acid sequence comprising a 5’ replacement construct and further comprising the sequence of one or more disclosed blocking U7 snRNAs.

[0324] Disclosed herein is a pharmaceutical formulation comprising a vector comprising a disclosed nucleic acid sequence comprising the sequence of one or more disclosed blocking U7snRNAs. Disclosed herein is a pharmaceutical formulation comprising a vector comprising a disclosed 3’ replacement construct that further comprises the sequence of one or more disclosed blocking U7 snRNAs. Disclosed herein is a pharmaceutical formulation comprising a vector comprising a disclosed nucleic acid sequence comprising a 3’ replacement construct and further comprising the sequence of one or more disclosed blocking U7 snRNAs.6. Transcriptome Engineering Systems

[0325] Disclosed herein is a transcriptome engineering system comprising one or more disclosed 3’ replacement constructs, one or more disclosed 5' replacement constructs, or any combination thereof. Disclosed herein is a transcriptome engineering system comprising one or more of 5" replacement constructs that enables the 5’ replacement of the targeted endogenous pre-mRNA. Disclosed herein is a transcriptome engineering system comprising one or more of 3’ replacement constructs that enables the 3’ replacement of the targeted endogenous pre-mRNA. Disclosed herein is a transcriptome engineering system comprising one or more of 5 ' replacement constructs that enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system comprising one or more of 3’ replacement constructs that enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more disclosed 3’ replacement constructs, wherein the nucleic acid molecule enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more disclosed 5’ replacement constructs, wherein the nucleic acid molecule enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA.

[0326] Disclosed herein is a transcriptome engineering system comprising one or more disclosed 3’ replacement constructs, one or more disclosed 5’ replacement constructs, or any combination thereof, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system comprising one or more of 5’ replacement constructs that enables the 5’ replacement of the targeted endogenous pre-mRNA, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system comprising one or more of 3’ replacement constructs that enable the 3‘ replacement of the targeted endogenous pre-mRNA, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system comprising one or more of 5’ replacement constructs that enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA, and one or more disclosed blocking U7 snRNAs.

[0327] Disclosed herein is a transcriptome engineering system comprising one or more of 3’ replacement constructs that enables the 3’ replacement of the targeted endogenous MYH7 premRNA, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more disclosed 3’ replacement constructs, wherein the nucleic acid molecule enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more disclosed 5’ replacement constructs, wherein the nucleic acid molecule enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA, and one or more disclosed blocking U7 snRNAs.

[0328] Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more disclosed 3’ replacement constructs, one or more disclosed 5’ replacement constructs, or any combination thereof. Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more 5’ replacement constructs that enables the 5’ replacement of the targeted endogenous pre-mRNA. Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more of 3’ replacement constructs that enables the 3’ replacement of the targeted endogenous pre-mRNA.

[0329] Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more of 5’ replacement constructs that enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more of 3’ replacement constructs that enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more vectors comprising one or more disclosed 3’ replacement constructs, wherein the nucleic acid molecule enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more vectors comprising one or more disclosed 5' replacement constructs, wherein the nucleic acid molecule enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA.

[0330] Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more disclosed 3’ replacement constructs, one or more disclosed 5’ replacement constructs, or any combination thereof, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more of 5’ replacement constructs that enables the 5’ replacement of the targeted endogenous pre-mRNA, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one ormore of 3’ replacement constructs that enables the 3’ replacement of the targeted endogenous pre-mRNA, and one or more disclosed blocking U7 snRNAs.

[0331] Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more of 5’ replacement constructs that enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system comprising one or more vectors comprising one or more of 3’ replacement constructs that enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more vectors comprising one or more disclosed 3’ replacement constructs, wherein the nucleic acid molecule enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more vectors comprising one or more disclosed 5’ replacement constructs, wherein the nucleic acid molecule enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA, and one or more disclosed blocking U7 snRNAs.

[0332] Disclosed herein is a transcriptome engineering system comprising one or more pharmaceutical formulations one or more disclosed 3’ replacement constructs, one or more disclosed 5’ replacement constructs, or any combination thereof. Disclosed herein is a transcriptome engineering system comprising one or more pharmaceutical formulations comprising one or more of 5 ' replacement constructs that enable the 5 ' replacement of the targeted endogenous pre-mRNA. Disclosed herein is a transcriptome engineering system comprising one or more pharmaceutical formulations compnsing one or more of 3’ replacement constructs that enable the 3’ replacement of the targeted endogenous pre-mRNA.

[0333] Disclosed herein is a transcriptome engineering system comprising one or more pharmaceutical formulations comprising one or more of 5 ’ replacement constructs that enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system comprising one or more pharmaceutical formulations comprising one or more 3’ replacement constructs that enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more pharmaceutical formulations comprising one or more disclosed 3’ replacement constructs, wherein the nucleic acid molecule enables the 3’ replacement of the targeted endogenous MYH7 pre-mRNA. Disclosed herein is a transcriptome engineering system for a targeted MYH7 pre-mRNA having one or more mutations or aberrations comprising one or more pharmaceuticalformulations comprising one or more disclosed 5’ replacement constructs, wherein the nucleic acid molecule enables the 5’ replacement of the targeted endogenous MYH7 pre-mRNA.

[0334] Disclosed herein is a transcriptome engineering system comprising one or more pharmaceutical formulations comprising one or more disclosed 3' replacement constructs, one or more disclosed 5’ replacement constructs, or any combination thereof, and one or more disclosed blocking U7 snRNAs. Disclosed herein is a ...

Claims

VIII. CLAIMSWhat is claimed is:

1. A nucleic acid molecule, comprising:one or more RNA structures comprising the sequence of any one of SEQ ID NO: 97 - SEQ ID NO: 106;one or more guide RNA sequences;a 3 ’ hemi intron; andan exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA.

2. A nucleic acid molecule, comprising:an exogenous RNA to be trans-spliced to a targeted endogenous MYH7 pre-mRNA; a 5 ’ hemi intron;one or more guide RNA sequences;one or more RNA structures comprising the sequence of any one of SEQ ID NO: 97 - SEQ ID NO: 106.

3. The nucleic acid molecule of Claim 1 or Claim 2, wherein the one or more guide RNA sequences comprise the sequence set forth in any one of SEQ ID NO:01 - SEQ ID NO:29 or SEQ ID NO:30 - SEQ ID NO:43.

4. The nucleic acid molecule of Claim 1, wherein the 3’ portion of the targeted endogenous MYH7 pre-mRNA comprises one or more mutations or aberrations, and wherein the one or more mutations in one or more exons contribute to pathogenesis in one or more cells.

5. The nucleic acid molecule of or Claim 2, wherein the 5’ portion of the targeted endogenous MYH7 pre-mRNA comprises one or more mutations or aberrations, and wherein the one or more mutations in one or more exons contribute to pathogenesis in one or more cells.

6. The nucleic acid molecule of Claim 4 or Claim 5, wherein the one or more cells are in a subject.

7. The nucleic acid molecule of Claim 1, wherein the one or more guide RNA sequences are directed to the intron immediately 3 ’ to the exon of the targeted endogenous MYH7 pre- mRNA with which it is to be spliced.

8. The nucleic acid molecule of Claim 2, wherein one or more guide RNA sequences are directed to the intron immediately 5’ to the exon of the targeted endogenous MYH7 pre-mRNA with which it is to be spliced.

9. The nucleic acid molecule of Claim 1, wherein the 3' hemi intron comprises the sequence set forth in SEQ ID NO: 90.

10. The nucleic acid molecule of Claim 1, wherein the 3’ hemi intron recruits a U2 snRNP, and wherein the U2 snRNP enables the formation of the spliceosome between the exogenous RNA to be trans-spliced and the targeted endogenous MYH7 pre-mRNA.

11. The nucleic acid molecule of Claim 2, wherein the 5’ hemi intron comprises the sequence set forth in SEQ ID NO: 91.

12. The nucleic acid molecule of Claim 2, wherein the 5' hemi intron recruits a U1 snRNP, and wherein the U 1 snRNP enables the formation of the spliceosome between the exogenous RNA to be trans-spliced and the targeted endogenous MYH7 pre-mRNA.

13. The nucleic acid molecule of Claim 1, comprising a sequence having at least 90% identity to the sequence set forth in SEQ ID NO: 86.

14. The nucleic acid molecule of Claim 2, comprising a sequence having at least 90% identity to the sequence set forth in SEQ ID NO: 89.

15. The nucleic acid molecule of any one of Claims 1 - 14, further comprising a promoter.

16. A viral vector, comprising: the nucleic acid molecule of any one of Claims 1 - 15.

17. A pharmaceutical formulation, comprising:the vector of Claim 16; andone or more pharmaceutically acceptable carriers, excipients, and / or binders.

18. A method of generating a chimeric RNA molecule, the method comprising:contacting a targeted endogenous pre-mRNA in one or more cells with the nucleic acid molecule of any one of Claim 1 - 15 or the vector of Claim 16, wherein the resulting chimeric RNA molecule comprises a trans-spliced nucleic acid sequence.

19. The method of Claim 18, wherein the one or more cells are in a subject.

20. The method of Claim 19, w herein the subj ect has one or more MYH7 mutations or aberrations.

21. A method of treating a subject, the method comprising:generating a chimeric RNA molecule in one or more cells by administering to a subject having one or more MYH7 mutations or aberrations a therapeutically effective amount of the vector of Claim 16 or the pharmaceutical formulation of Claim 17; wherein the resulting chimeric RNA molecule comprises a trans-spliced nucleic acid sequence; andwherein the resulting chimeric RNA molecule restores one or more aspects of cellular homeostasis and / or cellular functionality and / or metabolic dysregulation.

22. The method of Claim 21, wherein the therapeutically effective amount of the vector comprises about 1 x IO10vg to about 2 x 1014vg.

23. The method of Claim 21, wherein the 5’ portion of the targeted endogenous MYH7 pre-mRNA is trans-spliced with the exogenous RNA, or wherein a 3 ’ portion of the targeted MYH7 endogenous pre-mRNA is trans-spliced with the exogenous RNA.

24. The method of any one of Claims 21 - 23, further comprising administering to the subject one or more blocking U7 snRNAs, wherein the one or more blocking U7 snRNAs comprise the sequence set forth in any one of SEQ ID NO:62 - SEQ ID NO:71.

25. The method of any one of Claims 21 - 24, wherein, following the administering step, the health and / or wellness and / or functionality of the subject’s heart is improved or enhanced; and wherein the health and / or wellness and / or functionality of the subject’s heart is assessed by (i) measuring ejection fraction (EF), cardiac output (CO), stroke volume (SV), cardiac index (CI), wall motion / strain, or any combination thereof; and / or (ii) performing an echocardiogram, a cardiac MRI. an electrocardiogram, a nuclear medicine scan, or any combination thereof.

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