Compositions and methods for treating cep290-associated disease

Abstract

Compositions and methods for treatment of CEP290 related diseases are disclosed.

1 - 538 . (canceled) 
     
     
         539 . A dose for administration to a subject suffering from Leber’s Congenital Amaurosis 10 (LCA10) comprising
 an adeno-associated virus 5 (AAV5) vector capable of delivery to a non-dividing cell comprising in the same vector: 
 (i) a nucleotide sequence encoding a first gRNA molecule comprising a first targeting domain complementary with a first target domain from a CEP290 gene; 
 (ii) a nucleotide sequence encoding a second gRNA molecule comprising a second targeting domain complementary with a second target domain from the CEP290 gene; and 
 (iii) a nucleotide sequence encoding a Cas9 molecule; 
 wherein a AAV5 vector concentration in the dose is from 1x10 11  viral genomes (vg)/mL to 9x10 13  vg/mL. 
 
     
     
         540 . The dose of  claim 539 , wherein the concentration is selected from the group consisting of 1×10 11  vg/mL, 2×10 11  vg/mL, 3×10 11  vg/mL, 4×10 11  vg/mL, 5×10 11  vg/mL, 6×10 11  vg/mL, 7×10 11  vg/mL, 8×10 11  vg/mL, 9×10 11  vg/mL, 1×10 12  vg/mL, 2×10 12  vg/mL, 3×10 12  vg/mL, 4×10 12  vg/mL, 5×10 12  vg/mL, 6×10 12  vg/mL, 7×10 12  vg/mL, 8×10 12  vg/mL, 9×10 12  vg/mL, 1×10 13  vg/mL, 2×10 13  vg/mL, 3×10 13  vg/mL, 4×10 13  vg/mL, 5×10 13  vg/mL, 6×10 13  vg/mL, 7×10 13  vg/mL, 8×10 13  vg/mL, and 9×10 13  vg/mL. 
     
     
         541 . The dose of  claim 539 , wherein the first targeting domain comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:530 (CEP290-323), SEQ ID NO:555 (CEP290-485), SEQ ID NO:468 (CEP290-490), and SEQ ID NO:538 (CEP290-492) and the second targeting domain comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:558 (CEP290-64), SEQ ID NO:2321 (CEP290-11), SEQ ID NO:2312 (CEP290-230), SEQ ID NO:460 (CEP290-496), SEQ ID NO:586 (CEP290-502), and SEQ ID NO:568 (CEP290-504). 
     
     
         542 . The dose of  claim 540 , wherein the Cas9 molecule is an  S.   aureus  Cas9 molecule. 
     
     
         543 . The dose of  claim 541 , wherein in the first gRNA comprises a sequence having at least 90% sequence identity to a sequence selected from SEQ ID NOs: 2785 and 2787. 
     
     
         544 . The dose of  claim 541 , wherein the AAV5 vector encodes left and right ITRs having at least 90% sequence identity to SEQ ID NOs: 408 and 437. 
     
     
         545 . The dose of  claim 541 , wherein the dose is administered in a volume selected from the group consisting of at least 10 microliters, at least 50 microliters, at least 100 microliters, at least 150 microliters, at least 200 microliters, at least 250 microliters, and at least 300 microliters. 
     
     
         546 . A dose for administration to a subject suffering from Leber’s Congenital Amaurosis 10 (LCA10) comprising 
 an adeno-associated virus 5 (AAV5) recombinant viral particle capable of delivery to a non-dividing cell comprising: 
 (i) a nucleotide sequence encoding a first gRNA molecule comprising a first targeting domain complementary with a first target domain from a CEP290 gene; 
 (ii) a nucleotide sequence encoding a second gRNA molecule comprising a second targeting domain complementary with a second target domain from the CEP290 gene; and 
 (iii) a nucleotide sequence encoding a Cas9 molecule; 
 wherein a AAV5 recombinant viral particle concentration in the dose is from 1x10 11  viral genomes (vg)/mL to 9x10 13  vg/mL. 
 
     
     
         547 . The dose of  claim 546 , wherein the concentration is selected from the group consisting of 1×10 11  vg/mL, 2×10 11  vg/mL, 3×10 11  vg/mL, 4×10 11  vg/mL, 5×10 11  vg/mL, 6×10 11  vg/mL, 7×10 11  vg/mL, 8×10 11  vg/mL, 9×10 11  vg/mL, 1×10 12  vg/mL, 2×10 12  vg/mL, 3×10 12  vg/mL, 4×10 12  vg/mL, 5×10 12  vg/mL, 6×10 12  vg/mL, 7×10 12  vg/mL, 8×10 12  vg/mL, 9×10 12  vg/mL, 1×10 13  vg/mL, 2×10 13  vg/mL, 3×10 13  vg/mL, 4×10 13  vg/mL, 5×10 13  vg/mL, 6×10 13  vg/mL, 7×10 13  vg/mL, 8×10 13  vg/mL, and 9×10 13  vg/mL. 
     
     
         548 . The dose of  claim 546 , wherein the first targeting domain comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:530 (CEP290-323), SEQ ID NO:555 (CEP290-485), SEQ ID NO:468 (CEP290-490), and SEQ ID NO:538 (CEP290-492) and the second targeting domain comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:558 (CEP290-64), SEQ ID NO:2321 (CEP290-11), SEQ ID NO:2312 (CEP290-230), SEQ ID NO:460 (CEP290-496), SEQ ID NO:586 (CEP290-502), and SEQ ID NO:568 (CEP290-504). 
     
     
         549 . The dose of  claim 548 , wherein the Cas9 molecule is an  S.   aureus  Cas9 molecule. 
     
     
         550 . The dose of  claim 548 , wherein in the first gRNA comprises a sequence having at least 90% sequence identity to a sequence selected from SEQ ID NOs: 2785 and 2787. 
     
     
         551 . The dose of  claim 548 , wherein the AAV5 recombinant viral particle encodes left and right ITRs having at least 90% sequence identity to SEQ ID NOs: 408 and 437. 
     
     
         552 . The dose of  claim 548 , wherein the dose is administered in a volume selected from the group consisting of at least 10 microliters, at least 50 microliters, at least 100 microliters, at least 150 microliters, at least 200 microliters, at least 250 microliters, and at least 300 microliters. 
     
     
         553 . A dose or administration to a subject suffering from Leber’s Congenital Amaurosis 10 (LCA10) comprising 
 an adeno-associated virus 5 (AAV5) vector capable of delivery to a non-dividing cell comprising: 
 (i) a nucleotide sequence encoding a first gRNA molecule comprising a first targeting domain complementary with a first target domain from a CEP290 gene; and 
 (ii) 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 Cas9 molecule; 
 wherein a AAV5 vector concentration in the dose is from 1x10 11  viral genomes (vg)/mL to 9x10 13  vg/mL. 
 
     
     
         554 . The dose of  claim 553 , wherein the concentration is selected from the group consisting of 1×10 11  vg/mL, 2×10 11  vg/mL, 3×10 11  vg/mL, 4×10 11  vg/mL, 5×10 11  vg/mL, 6×10 11  vg/mL, 7×10 11  vg/mL, 8×10 11  vg/mL, 9×10 11  vg/mL, 1×10 12  vg/mL, 2×10 12  vg/mL, 3×10 12  vg/mL, 4×10 12  vg/mL, 5×10 12  vg/mL, 6×10 12  vg/mL, 7×10 12  vg/mL, 8×10 12  vg/mL, 9×10 12  vg/mL, 1×10 13  vg/mL, 2×10 13  vg/mL, 3×10 13  vg/mL, 4×10 13  vg/mL, 5×10 13  vg/mL, 6×10 13  vg/mL, 7×10 13  vg/mL, 8×10 13  vg/mL, and 9×10 13  vg/mL. 
     
     
         555 . The dose of  claim 553 , wherein the first targeting domain comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:530 (CEP290-323), SEQ ID NO:555 (CEP290-485), SEQ ID NO:468 (CEP290-490), and SEQ ID NO:538 (CEP290-492) and the second targeting domain comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:558 (CEP290-64), SEQ ID NO:2321 (CEP290-11), SEQ ID NO:2312 (CEP290-230), SEQ ID NO:460 (CEP290-496), SEQ ID NO:586 (CEP290-502), and SEQ ID NO:568 (CEP290-504). 
     
     
         556 . The dose of  claim 555 , wherein the Cas9 molecule is an  S.   aureus  Cas9 molecule. 
     
     
         557 . The dose of  claim 555 , wherein in the first gRNA comprises a sequence having at least 90% sequence identity to a sequence selected from SEQ ID NOs: 2785 and 2787. 
     
     
         558 . The dose of  claim 555 , wherein the dose is administered in a volume selected from the group consisting of at least 10 microliters, at least 50 microliters, at least 100 microliters, at least 150 microliters, at least 200 microliters, at least 250 microliters, and at least 300 microliters.