Large scale expression and purification of recombinant RNA in Escherichia coli

• Published in: Methods (San Diego, Calif.) Vol. 54; no. 2; pp. 267 - 273
• Main Authors: Ponchon, Luc, Dardel, Frédéric
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-06-2011
• Subjects: 5S Ribosomal RNA; Aptamer; Chromatography, Liquid; Cloning, Molecular; DNA, Recombinant - genetics; Escherichia coli; Escherichia coli - genetics; Expression plasmid; Genetic Vectors; Promoter Regions, Genetic; Recombinant RNA; RNA - genetics; RNA - isolation & purification; RNA - metabolism; Transcription, Genetic; tRNA; Aptamer; 5S Ribosomal RNA; Expression plasmid; tRNA; Recombinant RNA
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2011.02.007

Stable, folded RNA are involved in many key cellular processes and can be used as tools for biological, pharmacological and/or molecular design studies. However, their widespread use has been somewhat limited by their fragile nature and by the difficulties associated with their production on a large scale, which were limited to
in vitro methods. This work reviews the novel techniques recently developed that allow efficient expression of recombinant RNA
in vivo in
Escherichia coli. Based on the extensive data available on the genetic and metabolic mechanisms of this model organism, conditions for optimal production can be derived. Combined with a large repertoire of RNA motifs which can be assembled by recombinant DNA techniques, this opens the way to the modular design of RNA molecules with novel properties.