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Open Access Highly Accessed Research

Efficient inhibition of HIV-1 replication by an artificial polycistronic miRNA construct

Tao Zhang1, Tong Cheng1, Lihua Wei1, Yijun Cai1, Anthony Et Yeo1, Jiahuai Han2, Y Adam Yuan34, Jun Zhang1 and Ningshao Xia1*

  • * Corresponding author: Ningshao Xia sxia@xmu.edu.cn

  • † Equal contributors

Author Affiliations

1 National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Research Center for Medical Molecular Virology of Fujian Province, School of Life Science, Xiamen University, Xiamen, 361005, People’s Republic of China

2 The Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Science, Xiamen University, Xiamen, 361005, People’s Republic of China

3 Department of Biological Sciences and Temasek Life Sciences Laboratory, National University of Singapore, Singapore, 117543, Singapore

4 Xiamen-National University of Singapore Joint Laboratory in Biomedical Sciences, Xiamen University, Xiamen, 361005, People’s Republic of China

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Virology Journal 2012, 9:118  doi:10.1186/1743-422X-9-118

Published: 18 June 2012

Abstract

Background

RNA interference (RNAi) has been used as a promising approach to inhibit human immunodeficiency virus type 1 (HIV-1) replication for both in vitro and in vivo animal models. However, HIV-1 escape mutants after RNAi treatment have been reported. Expressing multiple small interfering RNAs (siRNAs) against conserved viral sequences can serve as a genetic barrier for viral escape, and optimization of the efficiency of this process was the aim of this study.

Results

An artificial polycistronic transcript driven by a CMV promoter was designed to inhibit HIV-1 replication. The artificial polycistronic transcript contained two pre-miR-30a backbones and one pre-miR-155 backbone, which are linked by a sequence derived from antisense RNA sequence targeting the HIV-1 env gene. Our results demonstrated that this artificial polycistronic transcript simultaneously expresses three anti-HIV siRNAs and efficiently inhibits HIV-1 replication. In addition, the biosafety of MT-4 cells expressing this polycistronic miRNA transcript was evaluated, and no apparent impacts on cell proliferation rate, interferon response, and interruption of native miRNA processing were observed.

Conclusions

The strategy described here to generate an artificial polycistronic transcript to inhibit viral replication provided an opportunity to select and optimize many factors to yield highly efficient constructs expressing multiple siRNAs against viral infection.

Keywords:
Artificial polycistronic transcript; HIV replication inhibition; Viral escape; RNA interference; siRNA