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US 9,023,649 B2Granted
RNA-guided human genome engineering
AssigneePresident and Fellows of Harvard College
InventorsPrashant G. Mali, George M. Church, Luhan Yang
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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.
Claims7
A method of altering a eukaryotic cell comprising
- providing to the eukaryotic cell guide RNA sequences complementary to target nucleic acid sequences,
- providing to the cell a Cas9 protein having inactive nuclease domains that interacts with the guide RNA sequence and binds to the target nucleic acid sequences in a site specific manner,
- wherein the Cas 9 has a FokI nuclease domain attached thereto,
- wherein a target nucleic acid sequence is cleaved upon FokI nuclease domain dimerization of adjacent guide RNA sequence and Cas9 protein co-localization complexes.
1 more element
Dependencies are inferred from the claim text.