Light activation of transcription: photocaging of nucleotides for control over RNA polymerization

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Abstract

We describe the use of ATP caged with [7-(diethylamino)coumarin-4-yl]methyl (DEACM) for light-controlled in vitro transcription reactions. Polymerization is blocked when DEACM is bonded to the gamma phosphate group of the ATP molecule. Controlled light irradiation releases ATP and transcription is initiated. In order to provide full control over the process, conditions involved in substrate release, nucleotide availability after release and the effect of the released coumarin in RNA polymerization were assessed in further detail. Together, our data provide the first direct evidence of control over enzymatic polymerization of nucleic acids through light. This approach may provide researchers with a unique tool for the study of biological processes at a molecular level.

Original languageEnglish
Article numbere90
JournalNucleic Acids Research
Volume36
Issue number14
DOIs
Publication statusPublished - 1 Aug 2008

Keywords

  • Photolysis
  • Light
  • Photolabile protecting

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