Nature Methods
- 4, 571 - 576 (2007)
Published online: 10 June 2007; | doi:10.1038/nmeth1058
Recombinant RNA technology: the tRNA scaffoldLuc Ponchon & Frédéric Dardel
Cristallographie & RMN Biologiques, Université Paris Descartes, CNRS, 4 avenue de l'Observatoire, 75006, Paris, France.
Correspondence should be addressed to Frédéric Dardel frederic.dardel@univ-paris5.fr 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.
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