US 12,286,623 B2
Methods and kits for identifying cancer treatment targets
Jennifer A. Doudna, Berkeley, CA (US); Chun-Hao Huang, Berkeley, CA (US); Spencer C. Knight, Berkeley, CA (US); and Nami Saghaei, Berkeley, CA (US)
Assigned to The Regents of the University of California, Oakland, CA (US)
Appl. No. 17/284,660
Filed by The Regents of the University of California, Oakland, CA (US)
PCT Filed Oct. 30, 2019, PCT No. PCT/US2019/058853
§ 371(c)(1), (2) Date Apr. 12, 2021,
PCT Pub. No. WO2020/092553, PCT Pub. Date May 7, 2020.
Claims priority of provisional application 62/753,631, filed on Oct. 31, 2018.
Prior Publication US 2021/0355487 A1, Nov. 18, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/10 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/90 (2006.01); G16B 30/10 (2019.01); G16B 40/00 (2019.01)
CPC C12N 15/1079 (2013.01) [C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/907 (2013.01); G16B 30/10 (2019.02); G16B 40/00 (2019.02); C12N 2310/20 (2017.05); C12N 2320/10 (2013.01); C12N 2800/80 (2013.01)] 21 Claims
 
1. A method comprising:
(a) introducing into a cancer cell and into a corresponding non-cancer cell of a same cell type as the cancer cell, a clustered regularly interspaced short palindromic repeat (CRISPR)/Cas system comprising:
i) a CRISPR/Cas effector polypeptide; and
ii) a CRISPR/Cas guide ribonucleic acid (RNA), or a nucleotide sequence encoding the CRISPR/Cas guide RNA, wherein the CRISPR/Cas guide RNA targets a target gene,
thereby producing a modified cancer cell and a modified non-cancer cell, both having reduced expression of the target gene;
(b) assaying RNA obtained from the modified cancer cell and the modified non-cancer cell to determine a molecular feature in the modified cancer cell and the modified non-cancer cell, thereby generating data associated with the molecular feature, wherein the molecular feature comprises an expression level of the target gene in the modified cancer cell and the modified non-cancer cell; and
(c) computer processing the data associated with the molecular feature to determine that the molecular feature is indicative of a reduction of cancerous state of the modified cancer cell,
wherein the determining comprises assessing an extent of transcriptional reversion toward a non-cancer cell expression level as a result of having reduced expression of the target gene.