RNA-guided human genome engineering
AssigneePresident and Fellows of Harvard College
InventorsGeorge M. Church, Prashant G. Mali, Luhan Yang
Life of this patent
Estimating the term…
Abstract
A method of altering a eukaryotic cell is provided including transfecting the eukaryotic cell with a nucleic acid encoding RNA complementary to genomic DNA of the eukaryotic cell, transfecting the eukaryotic cell with a nucleic acid encoding an enzyme that interacts with the RNA and cleaves the genomic DNA in a site specific manner, wherein the cell expresses the RNA and the enzyme, the RNA binds to complementary genomic DNA and the enzyme cleaves the genomic DNA in a site specific manner.
Claims18
A method of targeting a nuclease null Cas9 protein to a target nucleic acid sequence in a eukaryotic cell comprising
- providing the eukaryotic cell with a guide RNA sequence or a first nucleic acid sequence encoding the guide RNA sequence, wherein the guide RNA sequence comprises a spacer sequence and a scaffold sequence, wherein the spacer sequence is complementary to the target nucleic acid sequence,
- providing the eukaryotic cell with the nuclease null Cas9 protein, or a second nucleic acid sequence encoding the nuclease null Cas9 protein,
- wherein the guide RNA sequence and the nuclease null Cas9 protein form a colocalization complex at the target nucleic acid sequence, and
- wherein the guide RNA sequence is a crRNA-tracrRNA fusion transcript comprising GUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGA AAAAGUGGCACCGAGUCGGUGC (SEQ ID NO:46).
Dependencies are inferred from the claim text.