Recombinant RNA technology: the tRNA scaffold

被引:171
作者
Ponchon, Luc [1 ]
Dardel, Frederic [1 ]
机构
[1] Univ Paris 05, CNRS, F-75006 Paris, France
关键词
D O I
10.1038/NMETH1058
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:571 / 576
页数:6
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