The present invention relates to nucleic acid regulatory elements that are able to enhance muscle-specific expression of genes, in particular expression in cardiac muscle and/or skeletal muscle, methods employing these regulatory elements and uses of these elements. Expression cassettes and vectors containing these nucleic acid regulatory elements are also disclosed. The present invention is particularly useful for applications using gene therapy, more particularly muscle-directed gene therapy, and for vaccination purposes.

The present disclosure provides methods and compositions useful for the treatment or prevention of heart disease. In particular, the present disclosure provides a vector comprising a modified troponin promoter operatively linked to a therapeutic gene product for the treatment or prevention of heart disease, e.g., cardiomyopathy. The gene product may be MYBPC3. The disclosure also provides recombinant adeno-associated virus (rAAV) virions, rAAV viral genomes, and expression cassettes and pharmaceutical compositions thereof. The disclosure further provides methods for treating a disease or disorder, such as heart disease.

The present invention relates to methods for shifting the splicing profile of the DUX4 gene, a double homeobox gene on human chromosome 4q35. Recombinant adeno-associated viruses of the invention deliver DNAs encoding U7-based small nucle-ar RNAs to induce DUX4 exon- skipping and the expression of shortened forms of DUX4. The methods have application in the treatment of muscular dystrophies such as facioscapulohumeral muscular dystrophy.

Provided herein, in some embodiments, are methods and compositions for gene delivery. Provided herein is a technology for co-delivering to a cell (e.g., in vivo or ex vivo) enzymes capable of rearranging nucleic acid, such as site-specific recombinases, to directly assemble (e.g., covalently join) nucleic acid segments of, for example, a gene of interest.

A pharmaceutical composition can be provide for treating or preventing heart failure. Additionally, siRNA and a vector expressing said siRNA can be provided that can be used in the pharmaceutical composition for treating or preventing heart failure. For example, a pharmaceutical composition can be provided for treating or preventing heart failure that contains a DNA sequence encoding RNA containing a sense strand sequence of consecutive 18 to 29 nucleotides from angiopoietin-like protein 2 (ANGPTL2) mRNA or the alternative splicing type RNA thereof and an antisense strand sequence complementary to the sense strand sequence under control of a promoter, and a pharmaceutically acceptable carrier.

The present invention relates to a Crumbs homologue (CRB) therapeutic for use as a medicament or in a method of treatment or prophylaxis, for example in the treatment or prophylaxis of a retinal disorder due to mutations in the Crumbs homologue-1 (CRB1) gene, such as Leber's congenital amaurosis 8 (LCA8) or retinitis pigmentosa 12 (RP12). In particular, the present invention relates to a recombinant viral vector comprising CRB2 or modified non-toxic forms of either CRB1 or CRB3 that resemble CRB2.

The present invention relates to nucleic acid regulatory elements that are able to enhance muscle-specific expression of genes, in particular expression in cardiac muscle and/or skeletal muscle, methods employing these regulatory elements and uses of these elements. Expression cassettes and vectors containing these nucleic acid regulatory elements are also disclosed. The present invention is particularly useful for applications using gene therapy, more particularly muscle-directed gene therapy, and for vaccination purposes.

Disclosed are compositions and methods for cellular genome engineering that permit simple, efficient, and versatile permutations of combinatorial or simultaneous knockout and knock-in genomic modifications. An exemplary method includes modifying the genome of a cell by introducing to the cell a Cpf1 endonuclease and one or more AAV vectors encoding one or more crRNAs that direct the endonuclease to one or more target genes. The AAV vectors further contain one or more HDR templates that provide a sequence that encodes a reporter gene, a chimeric antigen receptor (CAR), or combinations thereof, and sequences homologous to one or more target sites. Also disclosed are pharmaceutical compositions containing genetically modified cells and methods of use thereof in treating a subject having a disease or disorder, such as cancer. The disclosed compositions and methods are especially applicable to development of enhanced chimeric antigen receptor engineered T cell therapy (CAR-T).

This disclosure provides gene therapy vectors comprising a polynucleotide encoding one or more isoforms of lysosome-associated membrane protein 2 (LAMP-2), and methods of using such gene therapy vectors for the treatment of Danon disease and other autophagy disorders.

Provided herein are variant adeno-associated virus (AAV) capsid proteins having one or more modifications in amino acid sequence relative to a parental AAV capsid protein, which, when present in an AAV virion, confer increased infectivity of one or more types of muscle cells as compared to the infectivity of the muscle cells by an AAV virion comprising the unmodified parental AAV capsid protein. Also provided are recombinant AAV virions and pharmaceutical compositions thereof comprising a variant AAV capsid protein as described herein, methods of making these rAAV capsid proteins and virions, and methods for using these rAAV capsid proteins and virions in research and in clinical practice, for example in, e.g., the delivery of nucleic acid sequences to one or more muscle cells for the treatment of muscle disorders and diseases.