Nondenaturing purification of co-transcriptionally folded RNA avoids common folding heterogeneity.

ABSTRACT: Due to the energetic frustration of RNA folding, tertiary structured RNA is typically characterized by a rugged folding free energy landscape where deep kinetic barriers separate numerous misfolded states from one or more native states. While most in vitro studies of RNA rely on (re)folding chemically and/or enzymatically synthesized RNA in its entirety, which frequently leads into kinetic traps, nature reduces the complexity of the RNA folding problem by segmental, co-transcriptional folding starting from the 5' end. We here have developed a simplified, general, nondenaturing purification protocol for RNA to ask whether avoiding denaturation of a co-transcriptionally folded RNA can reduce commonly observed in vitro folding heterogeneity. Our protocol bypasses the need for large-scale auxi

SUBMITTER: Pereira MJ 

PROVIDER: S-EPMC2944885 | biostudies-literature | 2010 Sep

REPOSITORIES: biostudies-literature