Biosynthetic studies of platensimycin

Kithsiri B. Herath, Athula B. Attygalle, Sheo B. Singh

Research output: Contribution to journalArticlepeer-review

60 Scopus citations

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 languageEnglish
Pages (from-to)15422-15423
Number of pages2
JournalJournal of the American Chemical Society
Volume129
Issue number50
DOIs
StatePublished - 19 Dec 2007

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