Cas9-mediated targeting of viral RNA in eukaryotic cells.
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.
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.
Aryn A
AA
Price
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.
Timothy R
TR
Sampson
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.
Hannah K
HK
Ratner
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.
Arash
A
Grakoui
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.
David S
DS
Weiss
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D000076987:CRISPR-Associated Protein 9; D064130:CRISPR-Associated Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry; D004720:Endonucleases / Q000378:metabolism*; D000939:Epitopes / Q000737:chemistry; D005603:Francisella / Q000378:metabolism*; D016174:Hepacivirus; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry*; D014162:Transfection; D014764:Viral Proteins / Q000378:metabolism
D020413:3' Untranslated Regions; D020121:5' Untranslated Regions; D001426:Bacterial Proteins; D064130:CRISPR-Associated Proteins; D000939:Epitopes; D012367:RNA, Viral; D014764:Viral Proteins; D004247:DNA; D000076987:CRISPR-Associated Protein 9; C000600049:Cas9 protein, Francisella novicida; D004720:Endonucleases
CRISPR-Cas; Cas9; Francisella; RNA targeting; hepatitis C virus