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Author Notes:

Email Address: david.weiss@emory.edu arash.grakoui@emory.edu

A.A.P., T.R.S., H.K.R., A.G., and D.S.W. designed research; A.A.P., T.R.S., and H.K.R. performed research.

A.A.P., T.R.S., H.K.R., A.G., and D.S.W. analyzed data; and A.A.P., T.R.S., H.K.R., A.G., and D.S.W. wrote the paper.

We thank Rafi Ahmed for helpful discussions, Sarkis Mazmanian for critical support, Yerkes Flow Cytometry Core for technical support, and Steven Bosinger of the Yerkes Genomics Core for performance of RNAseq and its analysis.

Conflict of interest statement: The authors have filed a related patent.

Subjects:

Research Funding:

This project was supported by National Institutes of Health (NIH) Grants R01-AI11070 and U54-AI057157 (Southeastern Regional Center of Excellence for Emerging Infections and Biodefense), the Burroughs Wellcome Fund (to D.S.W.), and Yerkes Research Center Base Grant P51RR-000165 and NIH Grants R01-AI070101 and R01-DK083356 (to A.G.).

T.R.S. was supported by the National Science Foundation Graduate Research Fellowship Program and the Achievement Rewards for College Scientists Foundation.

Keywords:

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

Cas9-mediated targeting of viral RNA in eukaryotic cells

Journal Title:

Proceedings of the National Academy of Sciences

Volume:

Volume 112, Number 19

Publisher:

, Pages 6164-6169

Type of Work:

Article | Final Publisher PDF

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.

Copyright information:

© 2015 National Academy of Sciences

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