The invention described herein provide methods and systems for deleting or inserting long (e.g., >100-500 bp) DNA sequences into a target DNA sequence using a single prime editing guide RNA (pegRNA) in conjunction with a CRISPR/Cas DNA nuclease.a

The present application relates to isolated novel CRISPR/Cas13 proteins, a gene editing systems based thereon, and a method for using said proteins for RNA level gene editing. Provided are non-naturally occurring or engineered RNA targeting systems, and said systems each have a novel Cas13 effector protein that targets RNA and at least one type of guide molecule.

Methods for editing the genome of cells such as T cells and hematopoietic stem cells are disclosed. The methods include inserting a nucleic acid sequence of an exogenous partial open reading frame (ORF) of an autosomal dominant gene (e.g., CTLA4) into an intronic target region of an endogenous autosomal dominant gene in the cell, wherein the endogenous autosomal dominant gene comprises one or more disease-causing mutations; the exogenous partial ORF of the autosomal dominant gene is free of disease-causing mutations; and insertion of the exogenous partial ORF of the autosomal dominant gene into the intronic target region results in a modified autosomal dominant gene that encodes a protein which is free of disease-causing mutations. Methods for treating haploinsufficiency and methods for increasing gene editing efficiency are also described.

Disclosed herein are gene editing systems comprising chemically modified RNAs (modRNAs) encoding Cas endonucleases or base editors. Wherein a gene editing system comprising a first chemically modified RNA (modRNA) comprising a sequence encoding a CRISPRassociated (Cas) endonuclease or a base editor; and a second modRNA comprising a sequence encoding a mutated p53 protein, wherein the mutated p53 protein inhibits a wild type p53 protein from binding to a target domain thereof.

Provided herein are natural killer (NK) cells with increased, modified or deleted extracellular matrix (ECM) receptors, and methods of making and using the same for cancer immunotherapy, and treatment of chronic infections, inflammation, autoimmune diseases, or transplant rejection.

Provided herein are compositions and methods related to modified prime editing guide RNAs.

An isolated immune cell, a method for preparing such modified immune cell, a method of treating a living being suffering or at risk of suffering from cancer or non-malignant diseases, an oligonucleotide and a use thereof.

Provided herein are compositions and methods for genetically engineering a cell (e.g., a hematopoietic cell) to modify a gene encoding a lineage-specific cell-surface antigen to modify an epitope of the lineage-specific cell-surface antigen recognized by an agent. Also provided are methods involving administering such genetically engineered cells to a subject, such as a subject having a hematopoietic malignancy, as well as the genetically engineered cells themselves.

The present invention includes systems, methods and compositions to modulate the IFN signaling pathway and its downstream effects on the innate immune response by regulating the expression of one or more IFNAR2 isoforms. In a preferred aspect, the modulation of the IFN signaling pathway may be accomplished by modulating the relative ration between IFNAR2 isoforms, IFNAR2-L and IFNAR2-S.

The present disclosure provides viral vectors which decrease undesired or off-target gene expression. Particularly, the present disclosure provides recombinant viral vectors comprising a transcription termination sequence adjacent to an inverted terminal repeat sequence (e.g., downstream of a first inverted terminal repeat sequence and upstream of an expression cassette), and virus or virus-like particles, compositions, and methods of using thereof.