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Recombinant RNA technology: the tRNA scaffold

Authors :
Frédéric Dardel
Luc Ponchon
Laboratoire de cristallographie et RMN biologiques (LCRB - UMR 8015)
Université Paris Descartes - Paris 5 (UPD5)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)
Université Paris Descartes - Paris 5 (UPD5) - Centre National de la Recherche Scientifique (CNRS)
Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)
Source :
Nature Methods, Nature Methods, Nature Publishing Group, 2007, 4 (7), pp.571-6. ⟨10.1038/nmeth1058⟩, Nature Methods, Nature Publishing Group, 2007, 4 (7), pp.571-6. 〈10.1038/nmeth1058〉
Publication Year :
2007

Abstract

International audience; RNA has emerged as a major player in most cellular processes. Understanding these processes at the molecular level requires homogeneous RNA samples for structural, biochemical and pharmacological studies. So far, this has been a bottleneck, as the only methods for producing such pure RNA have been in vitro syntheses. Here we describe a generic approach for expressing and purifying structured RNA in Escherichia coli, using tools that parallel those available for recombinant proteins. Our system is based on a camouflage strategy, the 'tRNA scaffold', in which the recombinant RNA is disguised as a natural RNA and thus hijacks the host machinery, escaping cellular RNases. This opens the way to large-scale structural and molecular investigations of RNA function.

Details

ISSN :
15487091 and 15487105
Volume :
4
Issue :
7
Database :
OpenAIRE
Journal :
Nature methods
Accession number :
edsair.doi.dedup.....de0a7614c6f621b0b778bc9d02f84529
Full Text :
https://doi.org/10.1038/nmeth1058⟩