Modified AAV constructs and uses thereof
In some aspects the disclosure relates to recombinant adeno-associated virus (rAAV) vectors and rAAVs (e.g., viral particles) engineered to express a transgene comprising an inhibitory nucleic acid (e.g., an artificial miRNA, amiRNA) having a pri-miRNA scaffold and a guide strand that targets a human target gene.
1. An rAAV vector comprising a transgene engineered to express an inhibitory nucleic acid comprising:
(i) a pri-miRNA scaffold; and
(ii) a guide strand that targets SOD1;
wherein the guide strand that targets SOD1 is encoded by an isolated nucleic acid comprising the sequence set forth in SEQ ID NO: 1.
2. The rAAV vector of claim 1 , wherein the pri-miRNA scaffold is selected from pri-miR-122, pri-miR-33, pri-miR-26a, pri-miR-126, pri-miR-22, pri-miR-199, pri-miR-99, pri-miR-21, pri-miR-375, pri-miR-101, pri-miR-451, pri-miR-194, pri-miR-30a, and pri-miR-155.
3. The rAAV vector of claim 1 , wherein the transgene comprises the sequence set forth in SEQ ID NO: 3.
4. The rAAV vector of claim 1 , wherein the rAAV vector is a self-complementary AAV (scAAV) vector.
5. An rAAV comprising the rAAV vector of claim 1 and an AAV capsid protein.
6. A method of reducing expression of SOD1 in a cell, the method comprising administering the rAAV of claim 5 to the cell.
7. A method for treating amyotrophic lateral sclerosis (ALS) in a subject in need thereof, the method comprising administering to the subject the rAAV of claim 5 .
8. The rAAV of claim 5 , wherein the AAV capsid protein is an AAV9 capsid protein.