title: Amplification of 4'-thioDNA and its utility as a shRNA expression system.
abstract: In this presentation, we discuss PCR amplification of 4'-thioDNA using 2'-deoxy-4'-thionucleoside triphosphate (sdNTPs). The amplified 4'-thioDNA acted as templates for not only in vitro transcription by T7 RNA polymerase, but also transcription in cells by RNA polymerase III. Accordingly, we succeeded to inhibit gene expression in cells by transfection of 4'-thioDNA that expresses a short-hairpin RNA (shRNA).
authors.affiliation: Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
authors.forename: Naonori
authors.initials: N
authors.lastname: Inoue
authors.forename: Aki
authors.initials: A
authors.lastname: Shionoya
authors.forename: Naoki
authors.initials: N
authors.lastname: Ogawa
authors.forename: Akira
authors.initials: A
authors.lastname: Matsuda
mesh_terms: D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D003854:Deoxyribonucleotides / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D022661:RNA, Untranslated / Q000096:biosynthesis* / Q000235:genetics; D013698:Templates, Genetic; D013873:Thionucleotides / Q000096:biosynthesis / Q000737:chemistry*; D014158:Transcription, Genetic
chemical_list: D003854:Deoxyribonucleotides; D022661:RNA, Untranslated; D013873:Thionucleotides; D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase