Recombinant RNA technology: the tRNA scaffold

Nat Methods. 2007 Jul;4(7):571-6. doi: 10.1038/nmeth1058. Epub 2007 Jun 10.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Escherichia coli / genetics
  • Humans
  • Nucleic Acid Amplification Techniques*
  • Nucleic Acid Conformation
  • RNA / biosynthesis*
  • RNA / chemistry
  • RNA / isolation & purification
  • RNA, Transfer / chemistry
  • RNA, Transfer / genetics*

Substances

  • RNA, recombinant
  • RNA
  • RNA, Transfer