Genetic screens and functional genomics using CRISPR/Cas9 technology.
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.
Department of Molecular and Cellular Biology, University of California Berkeley, Berkeley, CA, USA.
Ella
E
Hartenian
John G
JG
Doench
D064130:CRISPR-Associated Proteins / Q000502:physiology*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D005786:Gene Expression Regulation; D005818:Genetic Engineering / Q000379:methods*; D005820:Genetic Testing; D015894:Genome, Human; D006801:Humans; D017393:RNA Editing / Q000235:genetics*
D064130:CRISPR-Associated Proteins
CRISPR; Cas9; RNAi; functional genomics; gene editing; genetic screen; off-target effects; on-target activity; pooled screens; sgRNA