Real-time fluorescence detection of exoribonucleases

  1. Flore Sinturel1,
  2. Olivier Pellegrini1,
  3. Song Xiang2,
  4. Liang Tong2,
  5. Ciarán Condon1 and
  6. Lionel Bénard1
  1. 1Institut de Biologie Physico-Chimique, CNRS UPR 9073, Université de Paris 7–Denis Diderot, 75005 Paris, France
  2. 2Department of Biological Sciences, Columbia University, New York, New York 10027, USA

    Abstract

    The identification of RNases or RNase effectors is a continuous challenge, particularly given the current importance of RNAs in the control of genome expression. Here, we show that a fluorogenic RNA–DNA hybrid is a powerful tool for a real-time fluorescence detection and assay of exoribonucleases (RT-FeDEx). This RT-FeDEx assay provides a new strategy for the isolation, purification, and assay of known and unknown exoribonucleases.

    Keywords

    Footnotes

    • Reprint requests to: Lionel Bénard, Institut de Biologie Physico-Chimique, CNRS UPR 9073, Université de Paris 7–Denis Diderot, 13 rue Pierre et Marie Curie, 75005 Paris, France; e-mail: lionel.benard{at}ibpc.fr; fax: (33)-1-58-41-50-20.

    • Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.1670909.

      • Received March 31, 2009.
      • Accepted August 12, 2009.

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