title: Silencing of HIV-1 gene expression by two types of siRNA expression systems.
abstract: The RNA interference (RNAi) phenomenon is a recently discovered process in which the introduction of a double-stranded RNA (dsRNA) into cells causes the specific degradation of mRNA containing the same sequence. We designed mammalian expression vectors that direct the synthesis of small interfering RNA (siRNA)-like transcripts and examined them for their siRNA-mediated gene interference targeting the env gene (NL4-3:7490-7508, E7490). We constructed siRNA expression vectors for two different strands (sense and antisense; tandem promoter) and for siRNA expressed from the short hairpin RNA (shRNA). The inhibition efficacy on HIV-1 replication differed between these two vectors. Notably, the shRNA vector pU6-env-shRNA inhibited p24 production more effectively than the tandem promoter expression vector pU6-env-siRNA. Furthermore, we examined the ability of lentiviral vectors expressing shRNA to suppress HIV-1 expression in HIV-1-infected SupT1 cells. The env-shRNA (E 7490) almost completely suppressed HIV-1 expression in infected cells for up to 15 days.
authors.affiliation: Department of Life and Environmental Science, Chiba Institute of Technology, Narashino-shi, Chiba, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Hayafune
authors.forename: Naoko
authors.initials: N
authors.lastname: Miyano-Kurosaki
authors.forename: Wee-Sung
authors.initials: WS
authors.lastname: Park
authors.forename: Yuka
authors.initials: Y
authors.lastname: Moori
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Takaku
mesh_terms: D000818:Animals; D019380:Anti-HIV Agents / Q000494:pharmacology*; D019556:COS Cells; D002460:Cell Line; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D020868:Gene Silencing / Q000187:drug effects*; D005822:Genetic Vectors / Q000494:pharmacology; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics*; D006801:Humans; D008826:Microbial Sensitivity Tests; D034741:RNA, Small Interfering / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D014779:Virus Replication / Q000187:drug effects
chemical_list: D019380:Anti-HIV Agents; D034741:RNA, Small Interfering