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FEBS J. 2015 Apr;282(8):1383-93. doi: 10.1111/febs.13248. Epub 2015 Mar 16.

Genetic screens and functional genomics using CRISPR/Cas9 technology.

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Department of Molecular and Cellular Biology, University of California Berkeley, Berkeley, CA, USA.


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.


CRISPR; Cas9; RNAi; functional genomics; gene editing; genetic screen; off-target effects; on-target activity; pooled screens; sgRNA

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