Recombinant RNA technology: the tRNA scaffold

• Published in: Nature methods Vol. 4; no. 7; pp. 571 - 576
• Main Authors: Dardel, Frédéric, Ponchon, Luc
• Format: Journal Article
• Language: English
• Published: United States Nature Publishing Group 01-07-2007
• Subjects: Biochemistry; Biochemistry, Molecular Biology; Cell physiology; Cellular biology; E coli; Escherichia coli; Escherichia coli - genetics; Genetic aspects; Genetic recombination; Humans; Life Sciences; Nucleic Acid Amplification Techniques; Nucleic Acid Conformation; Physiological aspects; Properties; Recombinant proteins; Research methodology; Ribonucleic acid; RNA; RNA - biosynthesis; RNA - chemistry; RNA - isolation & purification; RNA, Transfer; RNA, Transfer - chemistry; RNA, Transfer - genetics; United States
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth1058

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.