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A fast, efficient and sequence-independent method for flexible multiple segmental isotope labeling of RNA using ribozyme and RNase H cleavage

Duss, Olivier ; Maris, Christophe ; von Schroetter, Christine ; Allain, Frédéric H.-T

In: Nucleic Acids Research, 2010, vol. 38, no. 20, p. e188-e188

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    Titre
    • A fast, efficient and sequence-independent method for flexible multiple segmental isotope labeling of RNA using ribozyme and RNase H cleavage
    Auteur
    • Duss, Olivier. Institute of Molecular Biology and Biophysics, ETH Zürich, CH-8093 Zürich, Switzerland
    • Maris, Christophe. Institute of Molecular Biology and Biophysics, ETH Zürich, CH-8093 Zürich, Switzerland
    • von Schroetter, Christine. Institute of Molecular Biology and Biophysics, ETH Zürich, CH-8093 Zürich, Switzerland
    • Allain, Frédéric H.-T. E-mail: allain@mol.biol.ethz.ch
    Type de document
    • Postprint
    Langue
    • Anglais
    Publié dans
    • Nucleic Acids Research, 2010, vol. 38, no. 20, p. e188-e188. Oxford University Press
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:300118
    Summary
    Structural information on RNA, emerging more and more as a major regulator in gene expression, dramatically lags behind compared with information on proteins. Although NMR spectroscopy has proven to be an excellent tool to solve RNA structures, it is hampered by the severe spectral resonances overlap found in RNA, limiting its use for large RNA molecules. Segmental isotope labeling of RNA or ligation of a chemically synthesized RNA containing modified nucleotides with an unmodified RNA fragment have proven to have high potential in overcoming current limitations in obtaining structural information on RNA. However, low yields, cumbersome preparations and sequence requirements have limited its broader application in structural biology. Here we present a fast and efficient approach to generate multiple segmentally labeled RNAs with virtually no sequence requirements with very high yields (up to 10-fold higher than previously reported). We expect this approach to open new avenues in structural biology of RNA