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RNA helicases play fundamental roles in modulating RNA structures and facilitating RNA–protein (RNP) complex assembly in vivo. Previously, our laboratory demonstrated that the DEAD-box RNA helicase Dbp2 in Saccharomyces cerevisiae is required to promote efficient assembly of the co-transcriptionally associated mRNA-binding proteins Yra1, Nab2, and Mex67 onto poly(A)+RNA. We also found that Yra1 associates...
Single-molecule fluorescence resonance energy transfer (smFRET) microscopy has become an increasingly popular tool to study the structural dynamics of RNA molecules. It reveals, in real time, the structural dynamics of these molecules that would be otherwise hidden in ensemble-averaged measurements. Here we present a detailed protocol for performing smFRET using total internal reflection fluorescence...
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