Efficient inhibition of HIV-1 replication by an artificial polycistronic miRNA construct.
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.
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.
Tao
T
Zhang
Tong
T
Cheng
Lihua
L
Wei
Yijun
Y
Cai
Anthony Et
AE
Yeo
Jiahuai
J
Han
Y Adam
YA
Yuan
Jun
J
Zhang
Ningshao
N
Xia
D000998:Antiviral Agents / Q000378:metabolism*; D001483:Base Sequence; D001688:Biological Products / Q000378:metabolism*; D002460:Cell Line; D015497:HIV-1 / Q000187:drug effects* / Q000502:physiology*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011995:Recombination, Genetic; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D001688:Biological Products; D035683:MicroRNAs