Monitoring Protein-RNA Interaction Dynamics In Vivo at High Temporal Resolution Using χCRACStuart W. McKellar 1, Ivayla Ivanova 1, Robert W. van Nues 2, Ross A. Cordiner 3, Mehak Chauhan 1, Niki Christopoulou 1, Will Worboys 4, Andrew Langford 4, Torben Heick Jensen 3, Sander Granneman 1
1Centre for Engineering Biology, University of Edinburgh, 2Institute of Cell Biology, University of Edinburgh, 3Department of Molecular Biology and Genetics, Aarhus University, 4UVO3 Ltd.
Kinetic cross-linking and analysis of cDNA is a method that allows investigation of the dynamics of protein-RNA interactions in living cells at high temporal resolution. Here the protocol is described in detail, including the growth of yeast cells, UV cross-linking, harvesting, protein purification, and next generation sequencing library preparation steps.
