Compositions and methods for treating CEP290-associated disease

A method of treating LCA10 in a subject in need thereof comprising administering to the subject:

• a recombinant viral particle comprising a nucleotide sequence encoding a first guide RNA (gRNA) molecule comprising a first targeting domain complementary with a first target domain from the CEP290 gene, a nucleotide sequence encoding a second gRNA molecule comprising a second targeting domain complementary with a second target domain from the CEP290 gene;
• and a nucleotide sequence encoding a Cas9 molecule;
• wherein the recombinant viral particle is administered to the eye of the subject, and
• wherein the recombinant viral particle is capable of delivery to a non-dividing cell, and wherein the administration results in NHEJ-mediated alteration of an LCA10 target position in one or more cells of the subject thereby treating the subject;
• and
• a corticosteroid administered daily at a first dose for at least 4 weeks beginning no more than fourteen days prior to administering the recombinant viral particle.

A method of treating LCA10 in a subject in need thereof comprising administering to the subject:

• an AAV vector comprising a nucleotide sequence encoding a first guide RNA (gRNA) molecule, a nucleotide sequence encoding a second gRNA molecule and a nucleotide sequence encoding a Cas9 molecule,
• wherein the AAV vector is administered to the eye of the subject,
• wherein the first and the second gRNA molecules are configured to form a first and a second complex with the Cas9 molecule, respectively, and
• wherein the first complex forms a first DNA double strand break between the 5′ end of the Alu repeat at c.2991+1162 to c.2991.1638 of intron 26 of the CEP290 gene and 500 nucleotides upstream of said 5′ end of the Alu repeat, and the second complex forms a second DNA double strand break between an LCA10 target position and 500 nucleotides downstream of the LCA10 target position, wherein the first and second DNA double strand breaks are repaired by NHEJ in a manner that results in alteration of the LCA10 target position thereby treating the subject;
• and
• a corticosteroid administered daily at a first dose for at least 4 weeks beginning no more than fourteen days prior to administering the AAV vector.