CPC C12N 15/907 (2013.01) [A01K 67/0275 (2013.01); A61K 48/0058 (2013.01); A61K 48/0091 (2013.01); C12N 7/00 (2013.01); C12N 9/22 (2013.01); C12N 15/1082 (2013.01); C12N 15/63 (2013.01); C12N 15/86 (2013.01); A01K 2217/052 (2013.01); A01K 2227/105 (2013.01); A01K 2267/0312 (2013.01); A01K 2267/0331 (2013.01); A01K 2267/0362 (2013.01); A61K 48/00 (2013.01); C12N 2710/24144 (2013.01); C12N 2740/15043 (2013.01); C12N 2750/14132 (2013.01); C12N 2750/14143 (2013.01); C12N 2750/14152 (2013.01); C12Y 301/00 (2013.01)] | 16 Claims |
1. A method of modifying a mammalian subject by editing in vivo a DNA target sequence in a genomic locus of interest of a hepatic cell in the mammalian subject, said method comprising in vivo delivering to the hepatic cell a single dose of a composition comprising a stable nucleic acid-lipid particle (SNALP) comprising a CRISPR-Cas system, wherein the CRISPR-Cas system comprises:
I. a CRISPR-Cas system RNA comprising a guide sequence that hybridizes to the DNA target sequence, and
II. a Cas9 comprising at least one nuclear localization sequence (NLS) or an mRNA encoding the Cas9,
wherein the guide sequence directs sequence-specific binding of a CRISPR complex to the DNA target sequence, the CRISPR complex comprises the Cas9 complexed with the CRISPR-Cas system RNA, and wherein the CRISPR complex introduces a double-stranded break in vivo in the DNA target sequence in the genomic locus of interest of the hepatic cell which forms a targeted indel in the genomic locus of interest and produces a phenotypic change in said mammalian subject, and wherein the delivering results in greater than 20% indel formation in the genomic locus of interest of the hepatic cell.
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