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US 11,078,498 B2Granted
CRISPR enabled multiplexed genome engineering
AssigneeThe Regents of the University of Colorado, a body corporate
InventorsAndrew Garst, Ryan T Gill
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Abstract
Described herein are method for generating a vector for editing a cell. The method comprises ligating into a vector that encodes a portion of a gRNA a cassette comprising at least one editing cassette, a promoter, and a gene encoding another portion of the gRNA. Upon ligation, the portion of the gRNA from the editing cassette and the other portion of the gRNA are ligated and form a functional gRNA.
Claims20
A method for generating a vector for editing a cell comprising:
- obtaining a vector that comprises a nucleotide sequence encoding a first region of a gRNA;
- and
- ligating into the vector that encodes the first region of the gRNA a second nucleotide sequence encoding:
- a region homologous to a target region in a cell comprising a change in sequence relative to the target region;
- a site conferring immunity to nuclease-mediated editing;
- a promoter;
- and
- a gene encoding a second region of the gRNA, whereupon the ligation forms a functional gRNA;
- thus generating the vector for editing the cell.
6 more elements
Dependencies are inferred from the claim text.