Multiplex RNA-guided genome engineering
AssigneePresident and Fellows of Harvard College
InventorsGeorge M. Church, James Dicarlo
Life of this patent
Estimating the term…
Abstract
Methods of multiplex genome engineering in cells using Cas9 is provided which includes a cycle of steps of introducing into the cell a first foreign nucleic acid encoding one or more RNAs complementary to the target DNA and which guide the enzyme to the target DNA, wherein the one or more RNAs and the enzyme are members of a co-localization complex for the target DNA, and introducing into the cell a second foreign nucleic acid encoding one or more donor nucleic acid sequences, and wherein the cycle is repeated a desired number of times to multiplex DNA engineering in cells.
Claims13
An isolated cell expressing an RNA guided DNA binding protein of a Type II CRISPR system that forms a co-localization complex with guide RNA complementary to target DNA and that cleaves the target DNA in a site specific manner comprising
- (a) a plurality of guide RNAs complementary to different sites of DNA within the cell, and
- (b) a plurality of exogenous double-stranded donor nucleic acid sequences, wherein each exogenous double-stranded donor nucleic acid sequence of the plurality of exogenous double-stranded nucleic acid sequences includes homology sequences or arms flanking a cut site.
Dependencies are inferred from the claim text.