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Nucleic Acids Res. 2017 Jul 27;45(13):7897-7908. doi: 10.1093/nar/gkx490.

Selective disruption of an oncogenic mutant allele by CRISPR/Cas9 induces efficient tumor regression.

Author information

1
Center for Genome Engineering, Institute for Basic Science (IBS), Seoul 08826, Korea.
2
Department of Basic Science, University of Science & Technology, Daejeon 34113, Korea.
3
Department of Bioengineering, College of Engineering, Hanyang University, Seoul 04763, Korea.
4
Department of Chemistry, Hanyang University, Seoul 04763, Korea.
5
Department of Chemistry, Seoul National University, Seoul 08826, Korea.

Abstract

Approximately 15% of non-small cell lung cancer cases are associated with a mutation in the epidermal growth factor receptor (EGFR) gene, which plays a critical role in tumor progression. With the goal of treating mutated EGFR-mediated lung cancer, we demonstrate the use of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) system to discriminate between the oncogenic mutant and wild-type EGFR alleles and eliminate the carcinogenic mutant EGFR allele with high accuracy. We targeted an EGFR oncogene harboring a single-nucleotide missense mutation (CTG > CGG) that generates a protospacer-adjacent motif sequence recognized by the CRISPR/Cas9 derived from Streptococcus pyogenes. Co-delivery of Cas9 and an EGFR mutation-specific single-guide RNA via adenovirus resulted in precise disruption at the oncogenic mutation site with high specificity. Furthermore, this CRISPR/Cas9-mediated mutant allele disruption led to significantly enhanced cancer cell killing and reduced tumor size in a xenograft mouse model of human lung cancer. Taken together, these results indicate that targeting an oncogenic mutation using CRISPR/Cas9 offers a powerful surgical strategy to disrupt oncogenic mutations to treat cancers; similar strategies could be used to treat other mutation-associated diseases.

PMID:
28575452
PMCID:
PMC5570104
DOI:
10.1093/nar/gkx490
[Indexed for MEDLINE]
Free PMC Article

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