In collaboration with Jankowsky and Licatalosi groups from the Center for RNA Science and Therapeutics and the Department of Biochemistry, members of the Crespo Group publish pioneering research on the dynamics of RNA-protein interactions in living cells. Proteins that act on RNA molecules rapidly associate and dissociate from their target binding sites, and measurements of the rates of association and dissociation are needed for a quantitative understanding of gene regulation. However, these measurements have been impossible to do in living cells. Writing in Nature, the authors now report on a technique they titled KIN-CLIP (for kinetics-crosslinking and immunoprecipitation) that enables the biochemical analysis of RNA-protein interactions in living cells for the first time. The authors make use of a pulsed femtosecond UV laser to crosslink proteins to RNA fast enough to capture their binding kinetics. The study brings the tools of physical chemistry and biochemistry into living cells, and, in doing so, provides an inflection point in the study of RNA–protein interactions. The KIN-CLIP method may also be used to investigate the crosslinking of proteins to DNA, as well as proteins to proteins, thus enabling the prospect of bringing it to bear on other types of interacting biomolecules in living cells.
Click here to view the article. Photo credit: Alexa Jankowsky