Exploiting the CRISPR/Cas9 PAM constraint for single-nucleotide resolution interventions

Yi Li, Saurabh Mendiratta, Kristina Ehrhardt, Neha Kashyap, Michael A. White, Leonidas Bleris

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

CRISPR/Cas9 is an enabling RNA-guided technology for genome targeting and engineering. An acute DNA binding constraint of the Cas9 protein is the Protospacer Adjacent Motif (PAM). Here we demonstrate that the PAM requirement can be exploited to specifically target single-nucleotide heterozygous mutations while exerting no aberrant effects on the wildtype alleles. Specifically, we 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 to a desired protospacer adjacent motif.

Original languageEnglish (US)
Article numbere0144970
JournalPLoS One
Volume11
Issue number1
DOIs
StatePublished - Jan 1 2016

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

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