Genome-wide mutagenesis screens for drug resistance in cancer.

The genetic mechansims of drug resistance elucidated to date from sequencing of pateint tumour samples following the development of resistance are neither exclusviely point mutations, gene amplifications or gene deletions but rather can encompass a combination of all of these. Thus the typical landscape of resistance effectors for any given drug tends to be composed of a number of genes that when mutated, amplified or deleted confer resistance. Therefore to systematically capture the breadth of resistance we plan to use genome-wide forward genetic screens - CRISPR/Cas9 (loss-of-function), CRISPR/SAM (gain-of-function) and chemical ENU mutagenesis (point mutations) - run in parallel on cancer cell lines to generate drug resistance and identify the causative genes by next-generation sequencing.