The Francis Crick Institute
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CRISPR-based large-scale modeling of loss-of-function mutations to investigate mechanisms of stress resistance in cancer

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journal contribution
posted on 2023-02-21, 11:14 authored by Fabrizio 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


Crick (Grant ID: 10152, Grant title: Scaffidi FC001152)