RNA-guided human genome engineering
First Claim
1. A method of altering a eukaryotic cell comprisingproviding the eukaryotic cell with a guide RNA having a spacer sequence complementary to genomic DNA of the eukaryotic cell and a scaffold sequence,providing the eukaryotic cell with a codon optimized Cas9 that interacts with the guide RNA and cleaves the genomic DNA in a site specific manner,wherein the guide RNA binds to complementary genomic DNA and the codon optimized Cas9 cleaves the genomic DNA in a site specific manner, and wherein the codon optimized Cas9 is a human codon optimized Cas9 encoded by a nucleic acid comprising SEQ ID NO:
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Accused Products
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.
47 Citations
12 Claims
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1. A method of altering a eukaryotic cell comprising
providing the eukaryotic cell with a guide RNA having a spacer sequence complementary to genomic DNA of the eukaryotic cell and a scaffold sequence, providing the eukaryotic cell with a codon optimized Cas9 that interacts with the guide RNA and cleaves the genomic DNA in a site specific manner, wherein the guide RNA binds to complementary genomic DNA and the codon optimized Cas9 cleaves the genomic DNA in a site specific manner, and wherein the codon optimized Cas9 is a human codon optimized Cas9 encoded by a nucleic acid comprising SEQ ID NO:
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7. A method of altering a eukaryotic cell at a plurality of genomic DNA sites comprising
providing the eukaryotic cell with a plurality of guide RNAs, each guide RNA having a spacer sequence complementary to a different site on genomic DNA of the eukaryotic cell and a scaffold sequence, providing the eukaryotic cell with a codon optimized Cas9, wherein the guide RNAs bind to complementary genomic DNA and the codon optimized Cas9 cleaves the genomic DNA in a site specific manner, and wherein the codon optimized Cas9 is a human codon optimized Cas9 encoded by a nucleic acid comprising SEQ ID NO:
Specification