Efficient incorporation of CoA, NAD and FAD into RNA by in vitro transcription

Nucleic Acids Res. 2003 Feb 1;31(3):e8. doi: 10.1093/nar/gng008.


Protein enzymes frequently recruit small molecule coenzymes to perform a variety of biochemical reactions. While the catalytic activities of RNA have been expanding rapidly, a similar strategy for RNA to utilize coenzymes and to increase its functional capabilities has yet to be demonstrated. A general in vitro transcription procedure has been developed to efficiently prepare RNA with coenzymes CoA, NAD and FAD covalently attached to the 5' end. These adenosine-containing coenzymes initiate transcription under the T7 class II promoter by T7 RNA polymerase. In addition to the three coenzymes, other adenosine-containing molecules may be incorporated into the first nucleotide position of RNA as well. This method provides easy access to CoA-, NAD- and FAD-RNA, which may find broad applications in generating coenzyme- utilizing ribozymes. In addition, both oxidized FAD and reduced NADH are highly fluorescent. NADH-RNA and FAD-RNA can therefore be used as probes for DNA/RNA detection and for structural investigation of RNA function by fluorescence spectroscopy.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adenosine / analogs & derivatives
  • Base Sequence
  • Coenzyme A / chemistry
  • Coenzyme A / metabolism*
  • Flavin-Adenine Dinucleotide / chemistry
  • Flavin-Adenine Dinucleotide / metabolism*
  • Molecular Sequence Data
  • NAD / chemistry
  • NAD / metabolism*
  • RNA / chemical synthesis*
  • RNA / chemistry
  • Spectrometry, Fluorescence
  • Transcription, Genetic*


  • NAD
  • Flavin-Adenine Dinucleotide
  • RNA
  • Adenosine
  • Coenzyme A