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Nucleic Acids Symposium Series 2007 51(1):57-58; doi:10.1093/nass/nrm029
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© 2007 Oxford University Press

Amplification of 4’-thioDNA and its utility as a shRNA expression system

Naonori Inoue1, Aki Shionoya2, Naoki Ogawa2 and Akira Matsuda1,2

1Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan and
2GeneticLab Co., Ltd., 2-12, Kita-27, Nishi-6, Kita-ku, Sapporo 001-0027, Japan

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).


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