title: Short communication: Efficiently inhibiting HIV-1 replication by a prototype foamy virus vector expressing novel H1 promoter-driven short hairpin RNAs.
abstract: RNA interference has shown great potential for the treatment of HIV-1. Vectors derived from prototype foamy viruses (PFVs) with a nonpathogenic nature are very promising gene transfer vehicles in anti-HIV gene therapy. In this article, three short hairpin RNAs (shRNAs) targeting the conserved regions of the HIV-1NL4-3 5' long terminal repeat (LTR) were first designed. We then constructed novel recombinant PFV vector plasmids, p▵Φ-H1-shRNAs, expressing these shRNAs under the control of the H1 RNA promoter. To detect the efficacy of these ▵Φ-H1-shRNAs for the inhibition of HIV-1 replication, we performed a dual-luciferase reporter assay, RT-qPCR, ELISA, western blotting, and a lactate dehydrogenase (LDH) assay by transient transfection in 293T cells. The results suggest that these novel shRNAs driven by PFV vectors inhibit HIV-1 replication efficiently without cytotoxicity, with shRNA3 being the most effective. In addition, we analyzed the shRNA target sites in the 5' LTR of HIV-1 strains other than HIV-1NL4-3 and found that these shRNAs may possibly inhibit other HIV-1 strains.
authors.affiliation: 1 Pathogenic Organism and Infectious Diseases Research Institute, School of Basic Medical Sciences, Wuhan University , Wuhan, China .
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Cheng
authors.forename: Lanlan
authors.initials: L
authors.lastname: Dong
authors.forename: Fengfeng
authors.initials: F
authors.lastname: Zhang
authors.forename: Peipei
authors.initials: P
authors.lastname: Yuan
authors.forename: Zhi
authors.initials: Z
authors.lastname: Li
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.forename: Jun
authors.initials: J
authors.lastname: Yin
authors.forename: Biwen
authors.initials: B
authors.lastname: Peng
authors.forename: Xiaohua
authors.initials: X
authors.lastname: He
authors.forename: Wanhong
authors.initials: W
authors.lastname: Liu
mesh_terms: D019380:Anti-HIV Agents / Q000378:metabolism*; D001688:Biological Products / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004337:Drug Carriers; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D020869:Gene Expression Profiling; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D016092:Spumavirus / Q000235:genetics; D013601:T-Lymphocytes / Q000502:physiology* / Q000821:virology*
chemical_list: D019380:Anti-HIV Agents; D001688:Biological Products; D004337:Drug Carriers; D034741:RNA, Small Interfering; D007770:L-Lactate Dehydrogenase