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Proc Natl Acad Sci U S A. 2015 May 12;112(19):6164-9. doi: 10.1073/pnas.1422340112. Epub 2015 Apr 27.

Cas9-mediated targeting of viral RNA in eukaryotic cells.

Author information

1
Department of Microbiology and Immunology, Microbiology and Molecular Genetics Program, Emory Vaccine Center, and Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329; and.
2
Emory Vaccine Center, and Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329; and Division of Infectious Diseases and david.weiss@emory.edu arash.grakoui@emory.edu.
3
Emory Vaccine Center, and Yerkes National Primate Research Center, Emory University, Atlanta, GA 30329; and Division of Infectious Diseases and Emory Antibiotic Resistance Center, Department of Medicine, Emory University School of Medicine, Atlanta, GA 30329 david.weiss@emory.edu arash.grakoui@emory.edu.

Abstract

Clustered, regularly interspaced, short palindromic repeats-CRISPR associated (CRISPR-Cas) systems are prokaryotic RNA-directed endonuclease machineries that act as an adaptive immune system against foreign genetic elements. Using small CRISPR RNAs that provide specificity, Cas proteins recognize and degrade nucleic acids. Our previous work demonstrated that the Cas9 endonuclease from Francisella novicida (FnCas9) is capable of targeting endogenous bacterial RNA. Here, we show that FnCas9 can be directed by an engineered RNA-targeting guide RNA to target and inhibit a human +ssRNA virus, hepatitis C virus, within eukaryotic cells. This work reveals a versatile and portable RNA-targeting system that can effectively function in eukaryotic cells and be programmed as an antiviral defense.

KEYWORDS:

CRISPR-Cas; Cas9; Francisella; RNA targeting; hepatitis C virus

PMID:
25918406
PMCID:
PMC4434742
DOI:
10.1073/pnas.1422340112
[Indexed for MEDLINE]
Free PMC Article

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