title: A fast, efficient and sequence-independent method for flexible multiple segmental isotope labeling of RNA using ribozyme and RNase H cleavage.
abstract: 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.
authors.affiliation: Institute of Molecular Biology and Biophysics, ETH Zürich, CH-8093 Zürich, Switzerland.
authors.forename: Olivier
authors.initials: O
authors.lastname: Duss
authors.forename: Christophe
authors.initials: C
authors.lastname: Maris
authors.forename: Christine
authors.initials: C
authors.lastname: von Schroetter
authors.forename: Frédéric H-T
authors.initials: FH
authors.lastname: Allain
mesh_terms: D002851:Chromatography, High Pressure Liquid; D011088:DNA Ligases; D007553:Isotope Labeling / Q000379:methods*; D019906:Nuclear Magnetic Resonance, Biomolecular; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D016337:RNA, Catalytic; D022661:RNA, Untranslated / Q000737:chemistry; D016914:Ribonuclease H; D014158:Transcription, Genetic
chemical_list: D016337:RNA, Catalytic; D022661:RNA, Untranslated; D012313:RNA; D016914:Ribonuclease H; D011088:DNA Ligases
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