US 12,486,518 B2
Bicistronic AAV vectors encoding hexosaminidase alpha and beta-subunits and uses thereof
Miguel Sena Esteves, Westford, MA (US); and Diane Golebiowski, Billerica, MA (US)
Assigned to University of Massachusetts, Westborough, MA (US)
Appl. No. 17/046,756
Filed by University of Massachusetts, Boston, MA (US)
PCT Filed Apr. 12, 2019, PCT No. PCT/US2019/027271
§ 371(c)(1), (2) Date Oct. 9, 2020,
PCT Pub. No. WO2019/200286, PCT Pub. Date Oct. 17, 2019.
Claims priority of provisional application 62/657,243, filed on Apr. 13, 2018.
Prior Publication US 2021/0095314 A1, Apr. 1, 2021
Int. Cl. C12N 15/86 (2006.01); A61K 48/00 (2006.01); A61P 25/00 (2006.01); C12N 9/24 (2006.01)
CPC C12N 15/86 (2013.01) [A61P 25/00 (2018.01); C12N 9/2402 (2013.01); C12Y 302/01052 (2013.01); A61K 48/00 (2013.01); C12N 2750/14143 (2013.01); C12N 2830/50 (2013.01)] 10 Claims
 
1. An isolated nucleic acid construct, comprising
(i) a first expression cassette, comprising a nucleic acid encoding a hexosaminidase alpha-subunit, and
(ii) a second expression cassette, comprising a nucleic acid encoding a hexosaminidase beta-subunit,
wherein the first expression cassette and the second expression cassette are operably linked by a bidirectional promoter, wherein the bidirectional promoter comprises two chicken 3-actin (CBA) promoters, and wherein the first expression cassette and the second expression cassette are flanked by adeno-associated virus (AAV) inverted terminal repeats (ITRs), and wherein the isolated nucleic acid construct comprises a sequence set forth in any one of SEQ ID NOs: 3-5 and 7-9.
 
8. An isolated nucleic acid construct, comprising
(i) a first expression cassette, comprising a nucleic acid encoding a hexosaminidase alpha-subunit, and
(ii) a second expression cassette, comprising a nucleic acid encoding a hexosaminidase beta-subunit,
wherein the first expression cassette and the second expression cassette are flanked by adeno-associated virus (AAV) inverted terminal repeats (ITRs), and wherein the isolated nucleic acid construct comprises the sequence set forth in SEQ ID NO: 6.