From Single Molecules to Live Cells: Imaging Nucleic Acid-Protein Interactions
13 May 2021
Prof. David Rueda
Groupleader at Department of Infectious Disease, Faculty of Medicine, Imperial College London, UK
"Research in the Rueda lab involves the development of quantitative single-molecule approaches to investigate the mechanism of complex biochemical systems (incl. RNA, DNA and protein). Single molecule microscopy has opened up new avenues leading to important discoveries on how structural dynamics correlate to the function of nucleic acids and proteins. An attractive aspect of single-molecule microscopy is that it reveals the structural dynamics of individual molecules, otherwise hidden in ensemble-averaged experiments, thereby providing direct observation of key reaction intermediates (even low populated or short lived ones) and the characterization of reaction mechanisms."
See more information at Prof. Rueda´s research group website.
About the lecture
Understanding how nucleic acids and proteins interact to regulate key cellular processes requires the ability to observe these interactions directly. The dynamic nature of many nucleic acid-protein interactions makes it challenging to study them with traditional bulk methods. Biochemical, molecular or cellular biology approaches yield ensemble- or population-averaged results, which may conceal key short-lived or low populated intermediates on the reaction pathway. To overcome the averaging problem, our group develops and applies single-molecule microscopy (SMM) approaches to monitor such interactions in real-time. SMM has become increasingly important in studies of nucleic acid-protein interactions because these techniques provide access to crucial information on how individual molecules or complexes behave in bulk solution and in live cells, revealing the underlying structural dynamics and heterogeneity in the system. We will present our data investigating some of these interactions on a specific model enzyme that plays key role in essential cellular processes and biotechnology: CRISPR-Cas9. We will also present our recent efforts to image RNA molecules in live and fixed mammalian cells with fluorogenic RNA aptamers (Mango).