RNA-Guided Human Genome Engineering
First Claim
1. A method of altering a eukaryotic cell comprisingproviding 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 protein 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.
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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.
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Citations
8 Claims
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1. 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 protein 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.
Specification