Compositions comprising nucleic acid molecules and methods of treating ATPase-mediated diseases
The present disclosure provides nucleic acid expression cassettes, vectors, compositions and methods for the treatment of ATPase-mediated diseases in a subject.
1. An isolated nucleic acid molecule, comprising from 5′ to 3′:
a first inverted terminal repeat sequence,
a human synapsin promoter sequence operably linked to a nucleic acid sequence encoding ATP1A3, wherein the nucleic acid sequence encoding ATP1A3 has at least 90% identity to the sequence set forth in SEQ ID NO:07, and
a second inverted terminal repeat sequence.
2. The isolated nucleic acid molecule of claim 1 , wherein the ATP1A3 comprises an amino acid sequence having at least 90% identity to SEQ ID NO:06.
3. The isolated nucleic acid molecule of claim 2 , wherein the ATP1A3 comprises the amino acid sequence of SEQ ID NO:06.
4. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid sequence encoding the ATP1A3 comprises the sequence set forth in SEQ ID NO:07.
5. The isolated nucleic acid molecule of claim 1 , wherein the nucleic acid sequence encoding the ATP1A3 is codon-optimized to reduce CpG methylation sites and for mammalian expression.
6. The isolated nucleic acid molecule of claim 1 , wherein the human synapsin promoter sequence comprises the nucleic acid sequence set forth in SEQ ID NO:05.
7. The isolated nucleic acid molecule of claim 1 , wherein the human synapsin promoter sequence comprises the nucleic acid sequence set forth in SEQ ID NO:09.
8. The isolated nucleic acid molecule of claim 1 , further comprising a bovine growth hormone polyadenylation signal (BGHpA).
9. The isolated nucleic acid molecule of claim 1 , comprising a nucleic acid sequence having at least 90% identity to the sequence set forth in SEQ ID NO:20.
10. The isolated nucleic acid molecule of claim 9 , comprising the nucleic acid sequence set forth in SEQ ID NO:20.
11. An adeno-associated virus (AAV) vector comprising the isolated nucleic acid molecule of claim 1 .
12. The AAV vector of claim 11 , wherein the AAV vector is AAV8.
13. The AAV vector of claim 11 , comprising the nucleic acid sequence set forth in SEQ ID NO:20.
14. A pharmaceutical composition comprising the AAV vector of claim 11 , and a pharmaceutically acceptable carrier and/or excipient.
15. The isolated nucleic acid molecule of claim 1 , further comprising one or more linker sequences.
16. The isolated nucleic acid molecule of claim 15 , wherein the one or more linker sequences comprise SEQ ID NO:11, SEQ ID NO:13, or SEQ ID NO:15.
17. The isolated nucleic acid molecule of claim 1 , further comprising a Simian virus 40 polyadenylation signal (SV40 pA) and/or a synthetic polyadenylation signal.