Optimized RPE65 promoter and coding sequences
The present invention relates to the prevention and/or treatment of retinal dystrophy in a patient, including Leber congenital amaurosis (LCA).
1. A method of treating retinal dystrophy caused by Retinal pigment epithelium-specific 65 kDa protein (RPE65) deficiency in a patient in need thereof, the method comprising administering to the patient via direct retinal, subretinal, or intravitreal injection, a therapeutically effective amount of an AAV vector comprising an expression construct comprising a promoter and an operably linked polynucleotide sequence, wherein the promoter consists of:
(a) nucleotides 12-761 of SEQ ID NO:2, or
(b) SEQ ID NO:2;
wherein the operably linked polynucleotide sequence comprises the sequence of SEQ ID NO: 4; and wherein expression of the operably linked polynucleotide sequence results in treatment of retinal dystrophy in the patient.
2. The method of claim 1 , wherein the promoter consists of nucleotides 12-761 of SEQ ID NO:2.
3. The method of claim 1 , wherein the promoter consists of SEQ ID NO: 2.
4. The method of claim 1 wherein the AAV vector comprises an AAV genome or a derivative thereof.
5. The method of claim 4 , wherein said derivative is a chimeric, shuffled or capsid modified derivative.
6. The method of claim 4 , wherein said AAV genome is from a naturally derived serotype or isolate or clade of AAV.
7. The method of claim 6 , wherein said AAV genome is from AAV serotype 2 (AAV2), AAV serotype 4 (AAV4), AAV serotype 5 (AAV5) or AAV serotype 8 (AAV8).
8. The method of claim 6 , wherein the vector comprises an AAV capsid wherein said capsid is derived from AAV5 or AAV8.
9. The method of claim 6 , wherein said AAV genome is from AAV serotype 2 (AAV2), AAV serotype 4 (AAV4), AAV serotype 5 (AAV5) or AAV serotype 8 (AAV8) and wherein the vector comprises an AAV capsid wherein said capsid is derived from AAV5 or AAV8.
10. The method of claim 9 , wherein the genome is derived from AAV2 and the capsid is derived from AAV5 or AAV8.
11. The method of claim 1 , wherein the retinal dystrophy is Leber congenital amaurosis (LCA).
12. The method of claim 1 , wherein administering the vector to the patient is performed by direct subretinal injection.