Transgene cassettes designed to express a human MECP2 gene
View Patent ↗The present disclosure provides methods and compositions for the treatment of diseases and genetic disorders linked to MeCP2 loss and/or misfunction, including RETT syndrome. The methods and compositions of the present disclosure comprise rAAV vectors and rAAV viral vectors comprising transgene nucleic acid molecules comprising nucleic acid sequences encoding for a MeCP2 polypeptide.
1. A method for treating Rett Syndrome in a subject in need thereof, comprising administering to the subject an rAAV viral vector, comprising (a) an AAV capsid protein and (b) an rAAV vector comprising, in 5′ to 3′ direction:
(i) a first AAV ITR sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 19;
(ii) a promoter sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 22;
(iii) a transgene comprising the nucleic acid sequence set forth in SEQ ID NO: 3;
(iv) a regulatory sequence comprising the nucleic acid sequence set forth in SEO ID NO: 13; and
(v) a second AAV ITR sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 21.
2. The method of claim 1 , wherein the rAAV vector comprises the nucleic sequence set forth in SEQ ID NO: 29.
3. The method of claim 1 , wherein the AAV capsid protein is an AAV9 capsid protein.
4. The method of claim 1 , wherein the rAAV viral vector is administered intrathecally.
5. The method of claim 1 , wherein at least about 10 14 viral particles are administered.
6. The method of claim 1 , wherein at least about 10 15 viral particles are administered.
7. The method of claim 1 , where in 10×10 14 viral particles are administered.
8. The method of claim 1 , wherein about 10 14 to about 10 15 viral particles are administered.
9. The method of claim 1 , wherein about 10 14 to about 10 15 viral particles are administered intrathecally.
10. The method of claim 1 , wherein the regulatory sequence further comprises an RDH1polyA element comprising the nucleic acid sequence set forth in SEQ ID NO: 14.
11. The method of claim 1 , wherein the regulatory sequence further comprises a MeCP2 downstream polyA element comprising the nucleic acid sequence set forth in SEQ ID NO: 16.
12. The method of claim 1 , wherein the regulatory sequence comprises the nucleic acid sequence set forth in SEQ ID NO: 17.
13. An rAAV vector, comprising, in 5′ to 3′ direction
(i) a first AAV ITR sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 19;
(ii) a promoter sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 22;
(iii) a transgene encoding a MECP2 polypeptide, the transgene comprising the nucleic acid sequence set forth in SEQ ID NO: 3;
(iv) a regulatory sequence comprising the nucleic acid sequence set forth in SEO ID NO: 13; and
(v) a second AAV ITR sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 21.
14. The rAAV vector of claim 13 , wherein the regulatory sequence further comprises an RDH1 polyA element comprising the nucleic acid sequence set forth in SEQ ID NO: 14.
15. The rAAV vector of claim 13 wherein the regulatory sequence further comprises a MeCP2 downstream polyA element comprising the nucleic acid sequence set forth in SEQ ID NO: 16.
16. The rAAV vector of claim 13 , wherein the regulatory sequence comprises the nucleic acid sequence set forth in SEQ ID NO: 17.
17. The rAAV vector of claim 13 , wherein the rAAV vector comprises the nucleic sequence set forth in SEQ ID NO: 29.
18. An rAAV viral vector comprising the rAAV vector of claim 13 and an AAV capsid protein.
19. The rAAV viral vector of claim 18 , wherein the AAV capsid protein is an AAV9 capsid protein.
20. A pharmaceutical composition comprising the rAAV viral vector of claim 19 .