	US 12,297,426 B2
	DNA damage response signature guided rational design of CRISPR-based systems and therapies
Uri Ben-David, Cambridge, MA (US); Todd Golub, Cambridge, MA (US); Rameen Beroukhim, Boston, MA (US); Oana Enache, Cambridge, MA (US); and Veronica Rendo, Boston, MA (US)
Assigned to THE BROAD INSTITUTE, INC., Cambridge, MA (US); and DANA-FARBER CANCER INSTITUTE, INC., Boston, MA (US)
Filed by THE BROAD INSTITUTE, INC., Cambridge, MA (US); and DANA-FARBER CANCER INSTITUTE, INC., Boston, MA (US)
Filed on Jan. 13, 2021, as Appl. No. 17/148,477.
Application 17/148,477 is a continuation of application No. 17/061,574, filed on Oct. 1, 2020, abandoned.
Claims priority of provisional application 62/909,131, filed on Oct. 1, 2019.
Prior Publication US 2021/0147828 A1, May 20, 2021
Int. Cl. C12N 15/10 (2006.01); A61K 35/13 (2015.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12Q 1/6883 (2018.01)

CPC C12N 15/1024 (2013.01) [A61K 35/13 (2013.01); C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12Q 1/6883 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01); C12Q 2600/156 (2013.01)]

22 Claims

1. A method for developing or designing a CRISPR-Cas based therapy or therapeutic comprising:

modifying one or more target sequence in an initial cell or initial cell population by expressing a Cas-protein and optionally a CRISPR-Cas system in the initial cell or initial cell population, thereby generating a modified cell or modified cell population;

clonally expanding the modified cell or modified cell population to obtain an expanded cell population;

detecting, in cells from the expanded cell population, expression of a Cas-induced DNA damage response protein signature; and

selecting clones from the expanded cell population that do not express the Cas-induced DNA-damage response signature and that do not have a p53 inactivating mutation.