title: Silencing of HIV-1 gag gene from epidemic strains among men who have sex with men (MSM) in Tianjin, China by a broad-spectrum short hairpin RNA.
abstract: RNA interference (RNAi) has been successfully used as a promising method to inhibit the replication of different viruses, including human immunodeficiency virus (HIV). Gene mutation is a hurdle for the anti-HIV by RNAi. Although prone to mutation, some genes are conserved and limited in functionally important regions. The gag gene is conserved in different subtypes and plays an important role in the assembly of HIV viral particle. Here, we identified subtypes and conserved sequences within forty-four gag genes from the epidemic strains among men who have sex with men. Three subtypes of gag gene, including CRF01_AE (47.7 %), CRF07_BC (40.9 %) and B (11.4 %) were analyzed by online blast. We designed five small hairpin RNAs (shRNAs) based on the conserved sequences. The gag-EGFP fusion transcript reporter system was used to select the most efficient shRNA. Among the five candidate shRNAs, gag-shRNA-3 represented a broad-spectrum inhibition against all chosen targets. This broad-spectrum shRNA diminished the titer of subtypes B and C of HIV-1 for a hundred orders of magnitude. The gag-shRNA-3 described here is a potential therapeutic agent in the HIV-1 gene therapy.
authors: School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, 300071 People's Republic of China.
authors: Zhipin
authors: Z
authors: Liang
authors: Tianjin Centers for Disease Control and Prevention, Tianjin, People's Republic of China.
authors: Xin
authors: X
authors: Wang
authors: School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, 300071 People's Republic of China.
authors: Hanmei
authors: H
authors: Li
authors: School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, 300071 People's Republic of China.
authors: Bin
authors: B
authors: Liu
authors: School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, 300071 People's Republic of China.
authors: Xuechao
authors: X
authors: Zhao
authors: School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, 300071 People's Republic of China.
authors: Chang
authors: C
authors: Liu
authors: School of Medicine, Nankai University, 94 Weijin Road, Nankai District, Tianjin, 300071 People's Republic of China.
authors: Xiao-Hong
authors: XH
authors: Kong
keywords: HIV-1; MSM; gag; shRNA