RNA-Guided Human Genome Engineering
First Claim
1. A method of modulating expression of a target nucleic acid in a cell comprisingproviding to the cell a guide RNA complementary to the target nucleic acid sequence,providing to the cell a Cas9 protein having inactive nuclease domains that interacts with the guide RNA and binds to the target nucleic acid sequence in a site specific manner,wherein the Cas 9 protein having inactive nuclease domains includes a transcriptional activator or repressor domain as a fusion protein for modulating target nucleic acid expression in vivo,wherein the guide RNA and the Cas9 protein including the transcriptional activator or repressor domain co-localize to the target nucleic acid sequence and wherein the transcriptional activator or repressor domain modulates expression of the target nucleic acid.
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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 modulating expression of a target nucleic acid in a cell comprising
providing to the cell a guide RNA complementary to the target nucleic acid sequence, providing to the cell a Cas9 protein having inactive nuclease domains that interacts with the guide RNA and binds to the target nucleic acid sequence in a site specific manner, wherein the Cas 9 protein having inactive nuclease domains includes a transcriptional activator or repressor domain as a fusion protein for modulating target nucleic acid expression in vivo, wherein the guide RNA and the Cas9 protein including the transcriptional activator or repressor domain co-localize to the target nucleic acid sequence and wherein the transcriptional activator or repressor domain modulates expression of the target nucleic acid.
Specification