US 12,122,997 B2
Excision of retroviral nucleic acid sequences
Kamel Khalili, Bala Cynwyd, PA (US); and Wenhui Hu, Cherry Hill, NJ (US)
Assigned to Temple University—of the Commonwealth System of Higher Education, Philadelphia, PA (US)
Appl. No. 15/998,558
Filed by Temple University—Of The Commonwealth System of Higher Education, Philadelphia, PA (US)
PCT Filed Feb. 13, 2017, PCT No. PCT/US2017/017652
§ 371(c)(1), (2) Date Aug. 15, 2018,
PCT Pub. No. WO2017/142835, PCT Pub. Date Aug. 24, 2017.
Claims priority of provisional application 62/410,496, filed on Oct. 20, 2016.
Claims priority of provisional application 62/363,625, filed on Jul. 18, 2016.
Claims priority of provisional application 62/345,520, filed on Jun. 3, 2016.
Claims priority of provisional application 62/337,994, filed on May 18, 2016.
Claims priority of provisional application 62/298,722, filed on Feb. 23, 2016.
Claims priority of provisional application 62/295,390, filed on Feb. 15, 2016.
Prior Publication US 2019/0225963 A1, Jul. 25, 2019
Int. Cl. C12N 15/11 (2006.01); A61K 48/00 (2006.01); C12N 9/22 (2006.01); C12N 15/113 (2010.01)
CPC C12N 15/11 (2013.01) [A61K 48/00 (2013.01); C12N 9/22 (2013.01); C12N 15/1132 (2013.01); A01K 2227/105 (2013.01); A01K 2267/0337 (2013.01); C07K 2319/09 (2013.01); C12N 2310/20 (2017.05); C12N 2750/14143 (2013.01)] 11 Claims
 
1. An adeno-associated viral vector serotype 9 (AAV9) expression vector, comprising a nucleic acid sequence encoding:
(a) a Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated endonuclease;
(b) a first guide RNA (gRNA), wherein the first gRNA targets a long terminal repeat (LTR) of a human immunodeficiency virus (HIV) sequence and the nucleic acid sequence encoding the first gRNA comprises any one of the rRNA encoding sequences of SEQ ID NOs: 1-6; and
(c) a second gRNA, wherein the nucleic acid sequence encoding the second gRNA comprises any one of the gRNA encoding sequences of SEQ ID NOs: 11 and 12,
wherein the CRISPR-associated endonuclease, the first gRNA, and the second gRNA are capable of excising intervening sequences between the first target sequence and the second target sequence.