Journal
FEBS JOURNAL
Volume 282, Issue 8, Pages 1383-1393Publisher
WILEY
DOI: 10.1111/febs.13248
Keywords
Cas9; CRISPR; functional genomics; gene editing; genetic screen; off-target effects; on-target activity; pooled screens; RNAi; sgRNA
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Functional genomics attempts to understand the genome by perturbing the flow of information from DNA to RNA to protein, in order to learn how gene dysfunction leads to disease. CRISPR/Cas9 technology is the newest tool in the geneticist's toolbox, allowing researchers to edit DNA with unprecedented ease, speed and accuracy, and representing a novel means to perform genome-wide genetic screens to discover gene function. In this review, we first summarize the discovery and characterization of CRISPR/Cas9, and then compare it to other genome engineering technologies. We discuss its initial use in screening applications, with a focus on optimizing on-target activity and minimizing off-target effects. Finally, we comment on future challenges and opportunities afforded by this technology.
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