Adeno-associated virus vectors encoding modified G6PC and uses thereof
View Patent ↗Modified G6PC (glucose-6-phosphatase, catalytic subunit) nucleic acids and glucose-6-phosphatase-α (G6Pase-α) enzymes with increased phosphohydrolase activity are described. Also described are vectors, such as adeno-associated virus (AAV) vectors, and recombinant AAV expressing modified G6Pase-α. The disclosed AAV vectors and rAAV can be used for gene therapy applications in the treatment of glycogen storage disease, particularly glycogen storage disease type Ia (GSD-Ia), and complications thereof.
1. An isolated nucleic acid molecule encoding a modified glucose-6-phosphatase-α (G6Pase-α), wherein the modified G6Pase-α comprises a serine to cysteine substitution at amino acid 298 of human G6Pase-α (SEQ ID NO: 2).
2. The isolated nucleic acid molecule of claim 1 , wherein the amino acid sequence of the modified G6Pase-α comprises or consists of SEQ ID NO: 8.
3. The isolated nucleic acid molecule of claim 1 , comprising the nucleotide sequence of SEQ ID NO: 6 or SEQ ID NO: 7.
4. A vector comprising the nucleic acid molecule of claim 1 .
5. The vector of claim 4 , wherein the nucleic acid molecule encoding the modified G6Pase-α is operably linked to a promoter.
6. The vector of claim 5 , wherein the promoter comprises a G6PC promoter.
7. The vector of claim 6 , wherein the G6PC promoter comprises nucleotides 182-3045 of SEQ ID NO: 4 or nucleotides 182-3045 of SEQ ID NO: 5.
8. The vector of claim 4 , comprising nucleotides 182-4441 of SEQ ID NO: 4 or nucleotides 182-4441 of SEQ ID NO: 5.
9. The vector of claim 4 , wherein the vector is an adeno-associated virus (AAV) vector.
10. The vector of claim 9 , wherein the AAV vector is an AAV serotype 8 (AAV8) vector.
11. The vector of claim 9 , comprising nucleotides 17-4819 of SEQ ID NO: 4 or nucleotides 17-4819 of SEQ ID NO: 5.
12. An isolated host cell comprising the nucleic acid molecule of claim 1 .