Patent ID: 11896651
Assignee: GENZYME CORPORATION
Field: Pharmaceuticals (Chemistry)
Classification: CPC A  C | IPC A  C

Claim 4:
5. A composition for treating Leber congenital amaurosis associated with a deep intronic mutation in a target DNA acid of an individual comprising:
a) a nucleic acid encoding an engineered, non-naturally occurring Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associated (Cas) (CRISPR-Cas) system comprising a first guide RNA and a second guide RNA, wherein the target DNA comprises a deep intronic mutation in a CEP290 gene and sequences flanking the mutation, wherein the first guide RNA and second guide RNA are capable of hybridizing to opposite strands of the flanking sequences; and
b) a recombinant adeno-associated virus (rAAV) vector comprising a Cas expression cassette, wherein the Cas expression cassette comprises:
i) a nucleotide sequence encoding a Cas9 protein operably linked to a promoter that is a single minimal CMV promoter consisting of nucleotides 34-216 of SEQ ID NO:56, and
ii) a target site for a single guide RNA, wherein the first guide RNA or the second guide RNA is capable of hybridizing to the target site;

wherein the Cas9 protein is capable of being expressed from the Cas expression cassette;
wherein the Cas9 protein is capable of cleaving the target DNA sequences flanking the mutation to thereby excise a portion of target DNA comprising the mutation; and
wherein the Cas9 protein is capable of cleaving the Cas expression cassette at the single target site in the Cas expression cassette to thereby reduce expression of the Cas9 protein, as compared to expression of the Cas9 protein prior to cleavage of the Cas expression cassette.