Abstract
Herein we describe the synthesis of 4′-thio-dTTP (1) and 4′-thio-dCTP (2) and their susceptibility for PCR amplification. Double stranded 4′-thioDNAs were amplified sufficiently by KOD dash DNA polymerase under appropriate conditions. Through this PCR study, it was shown that not only DNA-directed 4′-thioDNA synthesis but also 4′-thioDNA-directed 4′-thioDNA synthesis occurred in the presence of 1 and/or 2. We then tested the ability of the resulting 4′-thioDNAs to act as templates for transcription. In this experiment, the 4′-thioDNAs as templates afforded RNA in 38-49% yield relative to the case of the natural DNA template. We further investigated whether transcription occurred in mammalian cells. As a result, not only natural DNA but also 4′-thioDNAs were transcribed to afford a shRNA which showed gene-silencing effects via RNAi machinery.
Original language | English |
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Pages (from-to) | 15422-15423 |
Number of pages | 2 |
Journal | Journal of the American Chemical Society |
Volume | 129 |
Issue number | 50 |
DOIs | |
State | Published - 19 Dec 2007 |