Engineering of the Redox Ribozyme for the Determination of Its Architecture

Abstract

Here we report engineering efforts on an artificially evolved redox ribozyme, known as ribox02, and its more compact pseudoknot construct generated, combox02. The rigorous mutational works in the various regions suggested that the catalytically essential domain composed of three joining regions flanked by three stems. The architecture of combox02 can be an excellent framework for constructing libraries by randomizing sequences in this region for in vitro selection of novel ribozymes capable of catalyzing various chemical conversions of 5′-substrates.

An RNA aptamer that shifts the reduction potential of metabolic cofactors

“… 24 ). In vitro selection experiments identified an NAD + -dependent alcohol dehydrogenase ribozyme that uses NAD + to oxidize a benzylic alcohol to an aldehyde and performs the reverse reaction with the reduced form, NADH 25 – 27 . This ribozyme possesses the ability to alter the rate of the oxidation and reduction reactions, a kinetic property, but changes to the E m of the cofactor, a thermodynamic property, were not reported.…”