Provided herein are methods of delivering “split” Cas9 protein or nucleobase editors into a cell, e.g., via a recombinant adeno-associated virus (rAAV), to form a complete and functional Cas9 protein or nucleobase editor. The Cas9 protein or the nucleobase editor is split into two sections, each fused with one part of an intein system (e.g., intein-N and intein-C encoded by dnaEn and dnaEc, respectively). Upon co-expression, the two sections of the Cas9 protein or nucleobase editor are ligated together via intein-mediated protein splicing. Recombinant AAV vectors and particles for the delivery of the split Cas9 protein or nucleobase editor, and methods of using such AAV vectors and particles are also provided.

The disclosure relates, in some aspects, to compositions and methods for delivery of transgenes to a subject. In some embodiments, the disclosure provides expression constructs (e.g., vectors containing an expression construct) comprising a transgene encoding human Parkin or a portion thereof. In some embodiments, the disclosure provides methods of treating a neurodegenerative disease (e.g., Parkinson’s disease) by administering such expression constructs to a subject in need thereof.

The technology described herein is directed to adeno-associated vims (AAV) vectors comprising at least one gene regulatory element (GRE) and cells comprising said vectors. In another aspect, described herein are methods of screening for said gene regulatory elements. In another aspect, described herein are nucleic acid compositions comprising a GRE as described herein.

IPEX (Immune dysregulation Polyendocrinopathy X linked) syndrome is a primary immunodeficiency caused by mutations in the gene encoding the transcription factor forkhead box P3 (FOXP3), which leads to the loss of function of thymus-derived CD4+CD25+ regulatory T (tTreg) cells. Preclinical and clinical studies suggest that T cell gene therapy approaches designed to selectively restore the repertoire of Treg cells by transfer of wild type FOXP3 gene is a promising potential cure for IPEX. However, there is still a need for a vector that can be used efficiently for the preparation of said Treg cells. The inventors thus compared 6 different lentiviral constructs according to 4 criteria (vector titers, level of transduction of human CD4+ T cells, level of expression of FOXP3 and ΔLNGFR genes, degree of correlation between both expression) and selected one construct comprising a bidirectional PGK-EF1a promoter that showed remarkable efficiency.

Novel intron fragments are provided. The intron fragments can increase gene expression to levels equal to or higher than those achieved by the full-length intron while maintaining their ability to increase gene expression even when combined with various types of promoters and splicing donors. Particularly, the intron fragments enable loading of larger transgenes when used in genetic information delivery systems whose size is limited, for example, adeno-associated viruses (AAVs) and rhabdoviruses. Therefore, the use of the intron fragments is expected to extend the range of therapeutic genes.

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles.

Disclosed are a SARS-COV-2 antigen polypeptide, a recombinant adeno-associated virus (rAAV) expressing the polypeptide, and a vaccine containing the virus. A sequence of the antigen polypeptide is shown in SEQ NO. 1. and SEQ ID NO. 2. A method for preparing the recombinant adeno-associated virus comprises co-incubating pHelper, pRep2Cap5, and an expression vector, transfecting a cell in the presence of polyethyleneimine as a transfection reagent; culturing the cell, then collecting the cell by centrifugation, performing lysis and purification to obtain a purified liquid comprising the recombinant adeno-associated virus. The rAAV is delivered and expressed in vivo to produce a fusion antigen polypeptide, induces the production of serum neutralizing antibodies, which have a neutralizing titer to the novel SARS-COV-2 coronavirus and are expressed continuously; the rAAV composition can be used to immunize humans against the novel coronavirus pneumonia COVID-19.

Compositions and methods for the treatment of neurological disorders whereby vectors contain codon-optimized nucleic acids for expression in humans, encoding a human dopamine receptor D1 protein, a human 5-Hydroxytryptamine receptor 4 protein, and a human G-protein coupled receptor 139).

The present disclosure relates to one or more agents, therapies, treatments, and methods of use of the agents and/or therapies and/or treatments for increasing production of a belatacept-similar protein and interfering RNA of tumor necrosis factor alpha. Embodiments of the present disclosure can be used as a therapy or a treatment for a subject that has a condition whereby the subject's immune system is, or is likely to become, dysregulated and where the production of the belatacept-similar protein and decreased production of tumor necrosis factor alpha may be of therapeutic benefit.

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles.