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Identification of Transcription Factor Binding Sites by Cleavage Under Target and Release Using Nuclease in Zebrafish
Radwa Barakat1, Clyde A Campbell1, Raquel Espin-Palazon1
1Department of Genetics, Development and Cell Biology, Iowa State University, Ames, Iowa, USA.
Abstract:
Cleavage Under Targets and Release Using Nuclease (CUT&RUN) has emerged as a chromatin profiling strategy that excels traditional methods. Although CUT&RUN has been widely utilized in mammalian cells, its use in the zebrafish is at its early stages. In this study, we have developed a protocol to successfully perform CUT&RUN to map transcription factor (TF) binding sites in embryonic, adult tissues, and FACS-sorted zebrafish cells. We also provide a detailed workflow for the identification of predicted TF binding sites that can be utilized in any animal species. Altogether, our strategy will expand this invaluable tool to the zebrafish community, improving the epigenetic resolution that can be achieved in this model organism.

