TRANSGENE CASSETTES, AAV VECTORS AND AAV VIRAL VECTORS FOR THE EXPRESSION OF HUMAN CODON-OPTIMIZED SLC6A1
The present disclosure provides methods and compositions for the treatment of diseases and genetic disorders linked to SLC6A1 loss and/or misfunction. 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 GAT1 polypeptide.
1 . A method of treating epilepsy in a subject in need thereof, comprising administering to the subject an rAAV vector comprising, in 5′ to 3′ direction:
a) a first AAV ITR sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 12;
b) a promoter sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 14 or 15;
c) a transgene comprising the nucleic acid sequence set forth in any one of SEQ ID NOs: 3-10;
d) a polyA sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 16; and
e) a second AAV ITR sequence comprising the nucleic acid sequence set forth in SEQ ID NO: 13.
2 . The method of claim 1 , wherein the rAAV vector is a self-complementary AAV9 vector.
3 . The method of claim 1 , wherein the rAAV vector comprises the nucleic acid sequence set forth in SEQ ID NO: 20 or 22.
4 . The method of claim 1 , wherein the subject has a mutation in the SLC6A1 gene.
5 . The method of claim 1 , wherein the subject has SLC6A1 haploinsufficiency.
6 . The method of claim 1 , wherein the epilepsy is epilepsy with myoclonic-atonic seizures, genetic generalized epilepsy, or non-acquired focal epilepsy.
7 . The method of claim 1 , wherein the subject is less than 4 years of age.
8 . The method of claim 1 , wherein the rAAV viral vector is administered to the subject at a dose ranging from about 10 11 to about 1018 viral vector particles.
9 . The method of claim 1 , wherein the rAAV viral vector is administered intrathecally.