Mapping protein function with CRISPR/Cas9-mediated mutagenesis
Katherine F Donovan, Mudra HegdeiD, Meagan Sullender, Emma W VaimbergiD, Cory M Johannessen, David E Root, John G Doench
Abstract
CRISPR/Cas9 screening has proven to be a versatile tool for genomics research. We describe a CRISPR/Cas9-mediated approach to mutagenesis, exploiting the allelic diversity generated by error-prone non-homologous end-joining (NHEJ) to identify gain-of-function alleles of the MAPK signaling pathway genes MEK1 and BRAF. These results illustrate a scalable technique to easily generate cell populations containing thousands of endogenous allelic variants of any gene or genes to map variant functions.
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