posted on 2023-02-21, 11:14authored byFabrizio Simeoni, Ioannis Loukas, Thomas Stuart Wilson, Paola Scaffidi
Dissecting mechanisms driving subclone expansion in primary cancers has been challenging. Here, we present a protocol to systematically disrupt entire gene networks and assess the functional impact of this perturbation on cancer cell fitness. By combining arrayed CRISPR libraries and high-content microscopy, we describe steps to identify classes of genes whose inactivation promotes resistance to environmental challenges faced by cancer cells during tumor growth or upon therapy. A proof-of-principle interrogation of the epigenetic regulatory network is described.
For complete details on the use and execution of this protocol, please refer to Loukas et al. (2022).1
Funding
Crick (Grant ID: 10152, Grant title: Scaffidi FC001152)