Amplification of 4'-thioDNA and its utility as a shRNA expression system.
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).
Inoue
Naonori
N
Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan.
Shionoya
Aki
A
Ogawa
Naoki
N
Matsuda
Akira
A
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
D003854:Deoxyribonucleotides; D022661:RNA, Untranslated; D013873:Thionucleotides; D012321:DNA-Directed RNA Polymerases; D004259:DNA-Directed DNA Polymerase