(559f) Novel Approach to Target Endogenous KRAS Mutations By the Crispr/Cas9 Technology

Authors: 
Ehrhardt, K., The University of Texas at Dallas
Li, Y., The University of Texas at Dallas
Kashyap, N., The University of Texas at Dallas
Bleris, L., The University of Texas at Dallas

CRISPR/Cas9 is an enabling RNA-guided technology for genome targeting and engineering.  Here we demonstrate that the lentiviral-mediated CRISPR/Cas9 technology can be used to specifically target single-nucleotide heterozygous mutations while exerting no aberrant effects on the wild-type alleles. Specifically, we used a novel gRNA-constraint-mediated method to target the heterozygous G13A activating mutation of KRAS in colorectal cancer cells and we show reversal of drug resistance to a MEK small-molecule inhibitor. Our study introduces a new paradigm in genome editing and therapeutic targeting via the use of gRNA to guide Cas9.