Universal donor cells
AssigneeCrispr Therapeutics AG
InventorsAlireza Rezania, Rebeca Ramos-Zayas
Life of this patent
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Abstract
Genetically modified cells that are compatible with multiple subjects, e.g., universal donor cells, and methods of generating the genetically modified cells are provided herein. The universal donor cells comprise at least one genetic modification within or near at least one gene that encodes one or more MHC-I or MHC-II human leukocyte antigens or component or transcriptional regulator of the MHC-I or MHC-II complex, at least one genetic modification that increases the expression of at least one polynucleotide that encodes a tolerogenic factor, and optionally at least one genetic modification that increases or decreases the expression of at least one gene that encodes a survival factor.
Claims23
A method for generating a genetically modified cell, the method comprising delivering to a cell:
- (a) an RNA-guided nuclease;
- (b) a guide RNA targeting a target site in a beta-2-microglobulin (B2M) gene locus;
- and
- (c) a nucleic acid comprising (i) a nucleotide sequence homologous with a region located left of the target site in the B2M gene locus, (ii) a nucleotide sequence encoding a tolerogenic factor, and (iii) a nucleotide sequence homologous with a region located right of the target site in the B2M gene locus;
- wherein the B2M gene locus is cleaved at the target site and, through a process of homologous recombination, the nucleic acid of (c) is utilized as a template for inserting the nucleotide sequence encoding the tolerogenic factor into the B2M gene locus, thereby disrupting the B2M gene and generating a genetically modified cell, wherein the genetically modified cell expresses the tolerogenic factor and has disrupted expression of B2M.
Dependencies are inferred from the claim text.