US 11,913,017 B2
Efficient genetic screening method
Kun Zhang, La Jolla, CA (US); Prashant Mali, La Jolla, CA (US); Yan Wu, La Jolla, CA (US); and Dongxin Zhao, La Jolla, CA (US)
Assigned to The Regents of the University of California, Oakland, CA (US)
Appl. No. 16/312,907
Filed by The Regents of the University of California, Oakland, CA (US)
PCT Filed Jun. 28, 2017, PCT No. PCT/US2017/039825
§ 371(c)(1), (2) Date Dec. 21, 2018,
PCT Pub. No. WO2018/005691, PCT Pub. Date Jan. 4, 2018.
Claims priority of provisional application 62/356,438, filed on Jun. 29, 2016.
Prior Publication US 2019/0330661 A1, Oct. 31, 2019
Int. Cl. G16B 25/10 (2019.01); C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/113 (2010.01); C12N 15/90 (2006.01); G16B 20/20 (2019.01); G16B 40/10 (2019.01)
CPC C12N 15/907 (2013.01) [C12N 9/22 (2013.01); C12N 15/102 (2013.01); C12N 15/113 (2013.01); G16B 20/20 (2019.02); G16B 25/10 (2019.02); G16B 40/10 (2019.02); C12N 2310/20 (2017.05); C12N 2320/12 (2013.01); C12N 2330/31 (2013.01)] 19 Claims
 
1. A guide RNA comprising from 5′ to 3′:
(a) a gRNA spacer sequence at the 5′ end of the guide RNA, wherein the spacer sequence is complementary to a target gene,
(b) a scaffold sequence that binds to Cas9, and
(c) an RNA capture and sequencing domain comprising:
(1) a barcode sequence of 7-20 nucleotides, and
(2) a primer binding sequence comprising a polyA sequence of 20-25 adenosines.