Disclosed herein are nucleic acid-guided nucleases, guide nucleic acids, and targetable nuclease systems, and methods of use. Disclosed herein are engineered non-naturally occurring nucleic acid-guided nucleases, guide nucleic acids, and targetable nuclease systems, and methods of use. Targetable nuclease systems can be used to edit genetic targets, including recursive genetic engineering and trackable genetic engineering methods.

The present disclosure provides adeno-associated vims (AAV) vectors and uses thereof. In certain embodiments, the AAV vectors comprise a nucleic acid that encodes an antagonist against a family with sequence similarity 19, member A5 (FAM19A5) protein, e.g., anti-FAM19A5 antibody, e.g., anti-FAM19A5 scFv.

Disclosed herein are compositions and methods for the expression of a gene of interest. The disclosed methods may employ codon-optimization and introduction of non-endogenous restriction sites for efficient expression of a gene. The methods may further employ introduction of a gene variant of interest, such that the disclosed methods, compositions, and systems may be used to determine the significance of a variant of interest. Further disclosed are compositions, systems, and methods for the characterization of gene variants, and other mutations that may impact the function of the protein of interest.

The present invention discloses a CRISPR/LpCas9 gene editing system and application thereof, the CRISPR/LpCas9 gene editing system includes a complex of LpCas9 protein and sgRNA, which can accurately locate a target DNA sequence and cleave DNA double strands. The LpCas9 protein has an amino acid sequence shown in SEQ ID NO:1; and the sgRNA has a nucleotide sequence shown in SEQ ID NO:2, or a modified sgRNA sequence based on SEQ ID NO: 2. The present invention can effectively solve the problems of heterologous codon bias and cytotoxicity in the application of SpCas9 in Lactobacillus paracasei. The gene editing is performed in cells or in vitro by introducing artificially designed CRISPR RNA and a repair template, which solves the problem of low electroporation efficiency caused by the strains' characteristics or relatively large carried plasmids, and has broad application prospects in the field of gene editing.

Codon optimized nucleic acid sequences for RDH12 are provided, as well as recombinant viral vectors, such as AAV, expression cassettes, proviral plasmids or other plasmids containing the codon optimized sequence for functional RDH12. Recombinant vectors are provided that express the codon optimized, functional RDH12. Compositions containing these codon optimized sequences are useful in methods for treating, retarding or halting certain blinding diseases resulting from the absence, deficiency or inappropriate expression of RDH12. Other compositions and methods are providing for correcting a non-functional, defective or inadequately expressed native RDH12.

The present disclosure provides codon optimized nucleotide sequences encoding human alpha-galactosidase A, vectors, and host cells comprising codon optimized alpha-galactosidase A sequences, and methods of treating disorders such as Fabry disease comprising administering to the subject a codon optimized sequence encoding human alpha-galactosidase A.

The present invention relates to a nucleic acid molecule encoding human albumin for increasing the levels and/or activity of a protein or polypeptide encoded by a transgene, comprising a sequence defined by SEQ ID NO: 14 or a sequence having at least 80% sequence identity to said sequence, its use in nucleic acid expression cassettes and vectors containing liver-specific regulatory elements and codon-optimized factor IX, factor VIII, factor VII or factor VIIa transgenes, methods employing these expression cassettes and vectors and uses thereof. The present invention is particularly useful for applications using liver-directed gene therapy, in particular for the treatment of hemophilia A, hemophilia B or factor VII deficiency.

This invention relates to polynucleotides comprising optimized FIG4 open reading frame (ORF) sequences, vectors comprising the same, and methods of using the same for delivery of the ORF to a cell or a subject and to treat disorders associated with aberrant expression of a FIG4 gene or aberrant activity of a FIG4 gene product in the subject, such as CMT4J.

Respiratory syncytial virus (RSV)-based virus-like particles are disclosed. Also disclosed are polynucleotides encoding the virus-like particles (VLPs) as well as immunogenic compositions, pharmaceutical compositions, vaccines, and kits containing the virus-like particles. In addition, methods of producing and using each of the above compositions are also disclosed. Methods of use include single or combination administration of the RSV-VLPs, as well as use of the RSV-VLPs alone or in combination with other types of vaccines.

The present disclosure provides, among other things, a recombinant adeno-associated viral (rAAV) vector comprising a codon-optimized nucleotide sequence encoding an agent that inhibits the proteolytic activity of plasma kallikrein. The disclosure also provides, a recombinant adeno-associated viral (rAAV) vector encoding an anti-plasma kallikrein antibody heavy chain and an anti-plasma kallikrein antibody light chain.