Recombinant RNA technology: the tRNA scaffold.
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.
Ponchon
Luc
L
Cristallographie & RMN Biologiques, Université Paris Descartes, CNRS, 4 avenue de l'Observatoire, 75006, Paris, France.
Dardel
Frédéric
F
D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D009690:Nucleic Acid Conformation; D012313:RNA / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D012343:RNA, Transfer / Q000737:chemistry / Q000235:genetics*
C039919:RNA, recombinant; D012313:RNA; D012343:RNA, Transfer