Inhibition of HIV-1 Viral Infection by an Engineered CRISPR Csy4 RNA Endoribonuclease.
The bacterial defense system CRISPR (clustered regularly interspaced short palindromic repeats) has been explored as a powerful tool to edit genomic elements. In this study, we test the potential of CRISPR Csy4 RNA endoribonuclease for targeting HIV-1. We fused human codon-optimized Csy4 endoribonuclease with VPR, a HIV-1 viral preintegration complex protein. An HIV-1 cell model was modified to allow quantitative detection of active virus production. We found that the trans-expressing VPR-Csy4 almost completely blocked viral infection in two target cell lines (SupT1, Ghost). In the MAGI cell assay, where the HIV-1 LTR β-galactosidase is expressed under the control of the tat gene from an integrated provirus, VPR-Csy4 significantly blocked the activity of the provirus-activated HIV-1 reporter. This proof-of-concept study demonstrates that Csy4 endoribonuclease is a promising tool that could be tailored further to target HIV-1.
Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, China; Stanford University Medical School, Palo Alto, California, 94304, United States of America.
Rui
R
Guo
Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, China; Stanford University Medical School, Palo Alto, California, 94304, United States of America.
Hong
H
Wang
Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, China.
Jiuwei
J
Cui
Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, China.
Guanjun
G
Wang
Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, China.
Wei
W
Li
Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, China; Stanford University Medical School, Palo Alto, California, 94304, United States of America.
Ji-Fan
JF
Hu
D001426:Bacterial Proteins / Q000235:genetics*; D064130:CRISPR-Associated Proteins / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004722:Endoribonucleases / Q000235:genetics*; D057809:HEK293 Cells; D015658:HIV Infections / Q000517:prevention & control*; D019427:HIV Integrase / Q000502:physiology; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D015202:Protein Engineering; D019065:Virus Assembly; D016662:Virus Integration
D001426:Bacterial Proteins; D064130:CRISPR-Associated Proteins; D019427:HIV Integrase; C552242:Csy4 endoribonuclease, Pseudomonas aeruginosa; D004722:Endoribonucleases; C514801:p31 integrase protein, Human immunodeficiency virus 1