The present invention provides a complex containing a nucleic acid sequence-recognizing module and a proteolysis tag, wherein the module is linked to the proteolysis tag, the module specifically binds to a target nucleotide sequence in a double stranded DNA, and the tag consists of (i) a peptide containing 3 hydrophobic amino acid residues at the C-terminal, or (ii) a peptide containing 3 amino acid residues at the C-terminal wherein at least a part of the amino acid residues is substituted by serine

The present invention provides methods of producing genetically modified human hepatocytes suitable for hepatocyte transplantation comprising: disrupting one or more major histocompatibility complex (MHC) Class I or Class II genes in isolated human hepatocytes or in a hepatocyte progenitor cell by introducing a base editor and one or more gRNAs that hybridize with a target sequence in the one or more Class I or Class II genes, thereby producing genetically modified human hepatocytes.

Provided herein are systems, compositions, kits, and methods for the suppression of pain (e.g., chronic pain). Genes encoding ion channels (e.g., SCN9A) responsible for the propagation pain signals in neurons (e.g., DRG neurons) may be edited using a genome editing agent (e.g., a nucleobase editor). In some embodiments, loss-of-function ion channel mutants are generated, leading to pain suppression. In some embodiments, the genome editing agent is administered locally to the site of pain or to the nerves responsible for propagation of the pain signal.

Compositions and methods useful for expressing an RNA are provided. Compositions comprise nucleic acid molecules comprising an RNA polymerase III promoter and expression constructs comprising the novel promoters operably linked to a coding sequence. The promoters can initiate transcription of RNA-coding nucleotide sequences through the recruitment of RNA polymerase III (pol III) or, in some embodiments, polypeptide-encoding nucleotide sequences through the recruitment of RNA polymerase II. The promoters thus find use in transcribing nucleotide sequences, including but not limited to, those encoding guide RNAs, and polypeptide-encoding sequences. Such expression constructs comprising the novel promoters operably linked to nucleotide sequences encoding guide RNAs find use in binding and/or modifying the sequence of a target nucleic acid molecule, modifying the expression of a target gene, or detecting a target nucleotide sequence in cells comprising an RNA-guided nuclease (RGN) or RNA-guided nucleotide-binding polypeptide (RGNBP).

The technology described herein is directed to compositions, kits, systems and methods related to an engineered, inducible adenosine deaminase (iAD) enzymes, including but not limited to, an engineered inducible adenosine deaminase acting on RNA (ADAR) enzyme, which can be activated in the presence of an inducer. Also described are synthetic RNA molecules, to which the iAD can be specifically recruited to edit at least one target codon, leading to decreased or increased translation of the RNA molecules depending on the specific construct. The technology described herein is also directed to systems comprising the iAD and synthetic RNA molecule, nucleic acids and vectors encoding the iAD and synthetic RNA molecule, and methods of using such systems, nucleic acids, and vectors.

The present disclosure provides methods for treating cancer in a subject (by inhibiting e.g., APOBEC3A, APOBEC3B, or REV1), and methods of diagnosing cancer in a subject. Methods of tracking mutagenesis induced by a gene of interest (e.g., APOBEC3A, APOBEC3B, or REV1) and methods of screening for inhibitors and synthetic lethalities are also described herein. Further provided by the present disclosure are cell lines and antibodies for use in the methods described herein.

The present disclosure provides for endonuclease enzymes having distinguishing domain features, as well as methods of using such enzymes or variants thereof.

The present invention refers to a plant or plant part comprising a polynucleotide encoding a mutated TriA polypeptide, the expression of said polynucleotide confers to the plant or plant part tolerance to herbicides.

Some aspects of this disclosure provide compositions, methods, systems, and kits for controlling the activity of RNA-programmable endonucleases, such as Cas9, or for controlling the activity of proteins comprising a Cas9 variant fused to a functional effector domain, such as a nuclease, nickase, recombinase, deaminase, transcriptional activator, transcriptional repressor, or epigenetic modifying domain. For example, the inventive proteins provided comprise a ligand-dependent intein, the presence of which inhibits one or more activities of the protein (e.g., gRNA binding, enzymatic activity, target DNA binding). The binding of a ligand to the intein results in self-excision of the intein, restoring the activity of the protein.

Aspects of the disclosure relate to a gene therapy approach for diseases, disorders, or conditions caused by mutation in the stop codon utilizing modified tRNA. At least 10-15% of all genetic diseases, including muscular dystrophy (e.g. Duchene muscular dystrophy), some cancers, beta thalassemia, Hurler syndrome, and cystic fibrosis, fall into this category. Not to be bound by theory, it is believed that this approach is safer than CRISPR approaches due to minimal off-target effects and the lack of genome level changes.