Development of a microfluidic approach and its application to

In the last ten years the role of RNA in the regulation of gene expression has been shown to be of utmost importance. In order to study the kinetics of folding of regulatory RNAs (TPP riboswitches), we have developed a microfluidic approach.
The development of this platform is justified by two main advantages offered by this technology. First, we can reduce the volumes of reagents used, such as toxic probes (DMS) and/or of expensive probes (enzymatic probes). Second, the modularity of the technology will yield a greater kinetic-line complexity than that available with a conventional quench-flow device (four inputs maximum). It will also be possible to develop microfluidic platforms for the kinetic characterization of biological reactions that may involve a large number of partners.

Therefore, we have to transpose the quench-flow on a microfluidic chip containing several networks of microchannels for the flow of liquid. We have installed this microfluidic platform and we are engaged in the process of validating this kinetic approach. For that, we are using RNA footprinting by hydroxyl radicals which cleave the RNA chain following the accessibility of the hydrogen atoms of the ribose ring. The performance of the technology thus developed is estimated by comparing the results obtained by the conventional quench-flow device (kintek) and our newly developed microfluidic approach.