RNA-Guided Human Genome Engineering
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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
29 Claims
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1-14. -14. (canceled)
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15. A method of deleting a target nucleic acid sequence from a locus in a eukaryotic cell comprising
providing to the eukaryotic cell guide RNA sequences that are complementary to different target nucleic acid sequences, providing to the eukaryotic cell a Cas9 enzyme that interacts with the guide RNA sequences and cleaves the different target nucleic acid sequences, wherein the guide RNA sequences bind to the different target nucleic acid sequences and the Cas9 enzyme cleaves the different target nucleic acid sequences and the target nucleic acid sequence is deleted from the locus.
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22. A method of multiplexing genomic editing of target nucleic acids in a eukaryotic cell comprising
providing to the eukaryotic cell guide RNA sequences that are complementary to different target nucleic acid sequences, providing to the eukaryotic cell a Cas9 enzyme that interacts with the guide RNA sequences and cleaves the different target nucleic acid sequences, wherein the guide RNA sequences bind to the different target nucleic acid sequences and the Cas9 enzyme cleaves the different target nucleic acid sequences.
Specification