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US 2019/0376087 A1Published application
Crispr enabled multiplexed genome engineering
AssigneeThe Regents of the University of Colorado, a body corporate
InventorsAndrew GARST, Ryan T. GILL
Life of this patent
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Abstract
Described herein are methods and vectors for rational, multiplexed manipulation of chromosomes within open reading frames (e.g., in protein libraries) or any segment of a chromosome in a cell or population of cells, in which various CRISPR systems are used.
Claims20
A library of at least two vectors, each vector comprising:
- an inducible promoter driving transcription of the following covalently-linked elements:
- (i) a nucleic acid encoding a guide RNA (gRNA) comprising a region complementary to a target region of a nucleic acid in one or more cells;
- and (ii) an editing cassette comprising (a) a region homologous to the target region in the one or more cells with a mutation of at least one nucleotide relative to the target region in the one or more cells, and (b) a site conferring immunity to nuclease-mediated editing;
- wherein the at least two vectors have different mutations relative to the target region in the one or more cells.
1 more element
A library comprising a plurality vectors, wherein each vector comprises:
- an inducible promoter driving transcription of the following covalently-linked elements:
- (i) an editing cassette comprising:
- (a) a region homologous to a target region in one or more cells with a mutation of at least one nucleotide relative to the target region in the one or more cells, and (b) a site conferring immunity to nuclease-mediated editing;
- and (ii) a nucleic acid encoding a guide RNA (gRNA) complementary to the target region of the one or more cells, wherein the plurality of vectors have different mutations relative to the target region in the one or more cells.
1 more element
Dependencies are inferred from the claim text.