Figure 4.

An overview of the CRISPR approach for synthetic lethality screens. Matched cell lines stably expressing Cas9 are transduced with the gRNA library in a pooled virus format to produce a library of single cell mutants. The mutant library for each line is then screened for synthetic lethal interactions by isolating DNA and identifying underrepresented gRNA sequences in the cancer cell line, which are present in the matched (normal) cells. Interactions identified from the screen can be investigated and validated using RNAi. Small molecules interacting with the targets of interest can be developed and RNAi constructs may be used for in vivo studies.