7-Azidomethoxy-coumarins as profluorophores for templated nucleic acid detection.
Templated nucleic acid detection is an emerging bioanalytical method that makes use of the target DNA or RNA strand to initiate a fluorogenic reaction. The Staudinger reduction holds particular promise for templated sensing of nucleic acids because the involved functional groups are highly chemoselective. Here, the azidomethoxy group, which can be removed under Staudinger conditions, is used to cage 7-hydroxycoumarin fluorophores. Reduction by phosphines and subsequent loss of the azidomethoxy substituent induce a significant bathochromic shift of the major absorbance band in the near UV region. When excited at the appropriate wavelength, this change in the absorbance spectrum translates into a substantial fluorescence turn-on signal. The described profluorophores are readily conjugated to amino-modified DNAs and are rapidly uncaged by a triphenylphosphine-DNA probe under the control of a DNA template. In addition, turnover of the probes on the target strand occurs and yields substantial signal amplification.
Franzini
Raphael M
RM
Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA.
Kool
Eric T
ET
D003374:Coumarins / Q000138:chemical synthesis / Q000737:chemistry*; D004247:DNA / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D009696:Nucleic Acids / Q000032:analysis* / Q000378:metabolism; D012313:RNA / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D003374:Coumarins; D005456:Fluorescent Dyes; D009696:Nucleic Acids; D012313:RNA; D004247:DNA