The invention provides engineered RNA precursors that when expressed in a cell are processed by the cell to produce targeted small interfering RNAs (siRNAs) that selectively silence targeted genes (by cleaving specific mRNAs) using the cell's own RNA interference (RNAi) pathway. By introducing nucleic acid molecules that encode these engineered RNA precursors into cells in vivo with appropriate regulatory sequences, expression of the engineered RNA precursors can be selectively controlled both temporally and spatially, i.e., at particular times and/or in particular tissues, organs, or cells.

Pharmaceutical compositions including a muscle-specific nuclease cassette, one or more guide RNA cassettes, and a delivery system for delivery of the muscle-specific nuclease cassette and the one or more gRNA cassettes are provided. The pharmaceutical composition may also include a mutation-corrected DNA template including a modification to be made in a target nucleic acid sequence. Methods for treating a subject having a muscular or neuromuscular disorder are also provided. The methods may include administering to the subject a therapeutically effective amount of the pharmaceutical composition. Methods of modifying or editing the sequence of a target nucleic acid sequence in a muscle cell are also provided. The methods may include contacting or transducing the muscle cell with a muscle-specific nuclease cassette and one or more gRNA cassettes.

A multi-targeted siRNA for treating cancers is disclosed. Specifically, an siRNA composition is provided, comprising: a first siRNA molecule that reduces the expression of the first target gene; optionally, a coding sequence targeting a peptide element; and optionally, a second siRNA molecule that reduces the expression of the second target gene, wherein the first target gene is selected from the group consisting of EGFR, KRAS, or a combination thereof, and the siRNA composition reduces the expression of two or more genes. The siRNA or vector provided can be directly injected into the body to treat cancers.

A multi-targeted siRNA for treating cancers is disclosed. Specifically, an siRNA composition is provided, comprising: a first siRNA molecule that reduces the expression of the first target gene; optionally, a coding sequence targeting a peptide element; and optionally, a second siRNA molecule that reduces the expression of the second target gene, wherein the first target gene is selected from the group consisting of EGFR, KRAS, or a combination thereof, and the siRNA composition reduces the expression of two or more genes. The siRNA or vector provided can be directly injected into the body to treat cancers.

A method of treating an inherited retinal disease (IRD) associated with a pathogenic point mutation in a mutant allele of an IRD-related gene in the retina or the retinal pigment epithelium (RPE) of a subject in need thereof includes base editing the pathogenic point mutation in the retinal cell or retinal pigment epithelium cell to correct the pathogenic mutation, generate a non-pathogenic point mutation, or modulate expression of an IRD-related gene and restore visual function of subject.

The present invention provides for a double stranded RNA comprising a first RNA sequence and a second RNA sequence wherein the first and second RNA sequence are substantially complementary, wherein the first RNA sequence has a sequence length of at least 19 nucleotides and is substantially complementary to SEQ ID NO. 1. Said double stranded RNA is for use in inducing RNAi against Huntingtin exon 1 sequences. The double stranded RNA of to the invention was capable of reducing neuronal cell death and huntingtin aggregates in an animal model.

An object of the present invention is to provide a method of conveniently producing a genetically modified non-human mammal with high efficiency using a CRISPR-Cas9 system and particularly a production method whereby gene knock-in can be achieved with high efficiency regardless of the gene size. The method of producing a genetically modified non-human mammal comprises introducing a Cas9 protein, a crRNA fragment comprising a nucleotide sequence complementary to a target DNA region, and a tracrRNA fragment into a non-human mammalian oocyte to genetically modify the target DNA.

The present disclosure is broadly concerned with the field of cancer immunotherapy. For example, the present invention generally relates to an immune cell comprising a genetically engineered antigen receptor that specifically binds to a target antigen and a genetic disruption agent that reduces or is capable of reducing the expression in the immune cell of a gene that weakens the function of the immune cell.

The invention described herein provides gene therapy vectors, such as adeno-associated virus (AAV) vectors, that co-express two or more GOIs. The vectors of the invention can be broadly used to treat a number of genetic disorders such as trinucleotide repeat expansion disorders.

Viral vectors, lentiviral particles, and modified cells are disclosed. They encode or express a small RNA capable of targeting the KIF11 gene. In embodiments, the viral vectors and lentiviral particles further comprise and a KIF11 gene whose non-coding region has been modified such that it is resistant to activity by the small RNA.

A nucleic acid molecule comprising a stem-loop structure, a nucleic acid complex comprising the nucleic acid molecule, a composition for delivering a target recognition sequence, and the composition comprising the nucleic acid complex. An artificial small ribonucleic acid of the stem-loop structure stably maintains single-strandedness of a target recognition sequence which interacts with a nucleic acid of interest for a gene of interest of a prokaryote, thereby providing a nucleic acid complex for effective silencing of the gene of interest.