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.
87 Citations
32 Claims
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1-14. -14. (canceled)
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15. A method of making an HLA homozygous cell line comprising
providing to a cell line a guide RNA sequence complementary to a target nucleic acid sequence, providing to the cell a Cas9 protein, providing to the cell an HLA allele, wherein the guide RNA sequence binds to the target nucleic acid sequence and the Cas9 cleaves the target nucleic acid sequence, and wherein the HLA allele is included in the target nucleic acid.
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24. A method of making a humanized animal breed comprising
providing to a cell of an animal a guide RNA sequence complementary to a target nucleic acid sequence, providing to the cell of the animal a Cas9 protein, providing to the cell of the animal an HLA allele, wherein the guide RNA sequence binds to the target nucleic acid sequence and the Cas9 cleaves the target nucleic acid sequence, and wherein the HLA allele is included in the target nucleic acid.
Specification