US 11,807,895 B2
High-throughput drug and genetic assays for cellular transformation
Asaf Rotem, Cambridge, MA (US); Kevin Struhl, Cambridge, MA (US); Paul Blainey, Cambridge, MA (US); and Liyi Xu, Cambridge, MA (US)
Assigned to THE BROAD INSTITUTE, INC., Cambridge, MA (US); DANA FARBER CANCER INSTITUTE, INC., Boston, MA (US); PRESIDENT AND FELLOWS OF HARVARD COLLEGE, Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Appl. No. 15/560,977
Filed by THE BROAD INSTITUTE, INC., Cambridge, MA (US); DANA-FARBER CANCER INSTITUTE, INC., Boston, MA (US); President and Fellows of Harvard College, Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
PCT Filed Mar. 24, 2016, PCT No. PCT/US2016/024059
§ 371(c)(1), (2) Date Sep. 22, 2017,
PCT Pub. No. WO2016/154459, PCT Pub. Date Sep. 29, 2016.
Claims priority of provisional application 62/137,693, filed on Mar. 24, 2015.
Prior Publication US 2018/0051319 A1, Feb. 22, 2018
Int. Cl. C12Q 1/68 (2018.01); G01N 33/50 (2006.01); G01N 33/574 (2006.01); C12M 1/26 (2006.01)
CPC C12Q 1/68 (2013.01) [G01N 33/5008 (2013.01); G01N 33/5011 (2013.01); G01N 33/574 (2013.01); C12M 33/00 (2013.01)] 12 Claims
 
1. A method for identifying an agent that inhibits cellular growth and/or viability of tumorigenic or transformed cells in combination with a genetic perturbation, the method comprising:
providing a cellular sample of transformed cells and/or cells obtained from the tumor of a subject, wherein the transformed cells and/or cells obtained from the tumor are adherent cells;
infecting the cells with a barcoded library of lentiviruses encoding one or more CRISPR genetic perturbations, wherein the barcode identifies the one or more CRISPR genetic perturbations;
culturing the cellular sample for at least 5 days under low attachment conditions comprising agitating the cells, wherein the conditions inhibit cell attachment to the culture surface;
contacting or incubating the cellular sample with the agent; and
identifying the one or more genetic perturbations that are depleted in the sample contacted with the agent as compared to a control not contacted with the agent, whereby the agent inhibits cellular growth and/or viability of tumorigenic or transformed cells in combination with the one or more depleted genetic perturbations.