Inhibition of HIV-1 Viral Infection by an Engineered CRISPR Csy4 RNA Endoribonuclease

Rui Guo; Hong Wang; Jiuwei Cui; Guanjun Wang; Wei Li; Ji-Fan Hu

doi:10.1371/journal.pone.0141335

Inhibition of HIV-1 Viral Infection by an Engineered CRISPR Csy4 RNA Endoribonuclease

PLoS One. 2015 Oct 23;10(10):e0141335. doi: 10.1371/journal.pone.0141335. eCollection 2015.

Authors

Rui Guo¹, Hong Wang¹, Jiuwei Cui², Guanjun Wang², Wei Li², Ji-Fan Hu¹

Affiliations

¹ Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, China; Stanford University Medical School, Palo Alto, California, 94304, United States of America.
² Stem Cell and Cancer Center, First Hospital, Jilin University, Changchun, China.

Abstract

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / genetics*
CRISPR-Associated Proteins / genetics*
Clustered Regularly Interspaced Short Palindromic Repeats
Endoribonucleases / genetics*
HEK293 Cells
HIV Infections / prevention & control*
HIV Integrase / physiology
HIV-1 / physiology*
Humans
Protein Engineering
Virus Assembly
Virus Integration

Substances

Bacterial Proteins
CRISPR-Associated Proteins
HIV Integrase
Csy4 endoribonuclease, Pseudomonas aeruginosa
Endoribonucleases
p31 integrase protein, Human immunodeficiency virus 1

Grants and funding

This work was supported by a California Institute for Regenerative Medicine (CIRM) grant (RT2-01942), a Jilin International Collaboration Grant (#20120720), the National Natural Science Foundation of China grant (#81272294, #31430021) to J.F.H.; the National Natural Science Foundation of China grant (#81071920, #81372835) and Jilin Science and Technique Program grant (11GG003) to W.L.; and a grant of the Key Project of Chinese Ministry of Education (#311015) to C.J. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.