Delivery and use of the CRISPR-CAS systems, vectors and compositions for hepatic targeting and therapy

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 Cas9 is S. aureus Cas9.

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 DNA target sequence is adjacent to a protospacer adjacent motif (PAM) and the PAM is 5′-NNGRR.