CRISPR-based large-scale modeling of loss-of-function mutations to investigate mechanisms of stress resistance in cancer
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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
Journal details
Journal STAR Protocols
Volume 4
Issue number 1
Pages 102097
Available online
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Publisher website (DOI) 10.1016/j.xpro.2023.102097
Europe PubMed Central 36853711
Pubmed 36853711
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