Compositions and methods for treating cep290 associated disease

A nucleic acid encoding a Cas9, a first gRNA and a second gRNA, each gRNA targeted to a CEP290 gene for use in treating a CEP290-associated disease.

The nucleic acid for the use according to any one of the preceding claims wherein the nucleic acid encodes S. aureus Cas9.

The nucleic acid for the use according to any one of the preceding claims, wherein the Cas9 comprises a nuclear localization signal.

The nucleic acid for the use according to any one of the preceding claims, wherein the Cas9 comprises a polyadenylation signal.

The nucleic acid for the use according to any one of the preceding claims, wherein the nucleic acid comprises a promoter sequence for driving expression of the Cas9.

The nucleic acid for the use according to any one of the preceding claims, wherein the nucleic acid is to be administered as an adeno-associated virus (AAV) vector comprising the nucleic acid.

The nucleic acid for the use according to any one of the preceding claims, wherein the subject is a human.

A nucleic acid encoding a Cas9, a first gRNA and a second gRNA, wherein the first gRNA comprises a targeting domain selected from the group consisting of SEQ ID NOS:

• 1-3 and the second gRNA comprises a targeting domain selected from the group consisting of SEQ ID NOS:
• 4-6.

A nucleic acid encoding a Cas9, a first gRNA and a second gRNA, wherein the first gRNA comprises a targeting domain selected from the group consisting of SEQ ID NOS:

• 1-3 and the second gRNA comprises a targeting domain selected from the group consisting of SEQ ID NOS:
• 4-6 for use in a method of altering a retinal cell, wherein the method comprises administering the nucleic acid to a retinal cell in a subject.

One or more recombinant viral vectors comprising one or more nucleic acids encoding a Cas9, a first gRNA and a second gRNA for use in a method of altering a nucleotide sequence of the CEP 290 target position,

• wherein the method comprises contacting one or more recombinant viral vectors to the retina of a subject,
• wherein (a) the first and second gRNAs are adapted to form first and second ribonucleoprotein complexes with the Cas9, and (b) the first and second ribonucleoprotein complexes are adapted to cleave first and second cellular nucleic acid sequences on first and second sides of a CEP290 target position, thereby altering a nucleotide sequence of the CEP290 target position.

A nucleic acid encoding a Cas9, a first gRNA and a second gRNA, wherein each of the first and second gRNAs comprise a sequence having at least 90% sequence identity to one of SEQ ID NOS:

• 7-8.

A nucleic acid encoding a Cas9, a first gRNA and a second gRNA, wherein each of the first and second gRNAs comprise a sequence selected from the group consisting of SEQ ID NOS:

• 7-8, or a sequence having at least 90% sequence identity thereto.

A method of treating a subject, comprising:

• administering to the subject a nucleic acid encoding a Cas9, a first gRNA and a second gRNA, each gRNA targeted to a CEP290 gene of the subject.

A nucleic acid encoding a Cas9, a first gRNA and a second gRNA, wherein the first gRNA comprises a targeting domain selected from the group consisting of SEQ ID NOS:

• 1-3 and the second gRNA comprises a targeting domain selected from the group consisting of SEQ ID NOS:
• 4-6.

A method of treating a subject, comprising:

• contacting a retina of the subject with one or more recombinant viral vectors comprising one or more nucleic acids encoding a Cas9, a first gRNA and a second gRNA,
• wherein (a) the first and second gRNAs are adapted to form first and second ribonucleoprotein complexes with the Cas9, and (b) the first and second ribonucleoprotein complexes are adapted to cleave first and second cellular nucleic acid sequences on first and second sides of a CEP290 target position, thereby altering a nucleotide sequence of the CEP290 target position.

A method of treating a subject having an inherited retinal dystrophy, comprising:

• contacting a retina of the subject with one or more recombinant viral vectors comprising one or more nucleic acids encoding a Cas9 and a first gRNA comprising a sequence having at least 90% sequence identity to a sequence selected from SEQ ID NOS:
• 7 and 8;
• wherein (a) the first gRNA is adapted to form a first ribonucleoprotein complex with the Cas9, and (b) the first ribonucleoprotein complex is adapted to cleave a first cellular nucleic acid sequence associated with the inherited retinal dystrophy, thereby altering the first cellular nucleic acid sequence.