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US 2019/0194693 A1Published application
Crispr enabled multiplexed genome engineering
AssigneeThe Regents of the University of Colorado, a body corporate
InventorsRyan T. Gill, Andrew Garst
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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 single vector comprising at least two synthetic oligonucleotides, wherein each synthetic oligonucleotide comprises the following covalently-linked components:
- (i) a nucleic acid encoding a gRNA sequence targeting a target region in a cell;
- (ii) a region homologous to the target region comprising a change in sequence relative to the target region;
- and (iii) a site conferring immunity to nuclease-mediated editing;
- and wherein the at least two synthetic oligonucleotides are homologous to different target regions in the cell.
1 more element
A single vector comprising at least two synthetic oligonucleotides, wherein each synthetic oligonucleotide comprises the following covalently-linked components:
- (i) a nucleic acid encoding a gRNA sequence targeting a target region in a cell;
- and (ii) a region homologous to the target region comprising a change in sequence relative to the target region, wherein the at least two synthetic oligonucleotides are homologous to different target regions in the cell.
Dependencies are inferred from the claim text.