High-efficiency genomic mapping of chromatin-associated targets with CUT&RUN
Firestone, T. M.; Venters, B. J.; Novitzky, K.; Albertorio-Saez, L. M.; Barnes, C. A.; Fedder-Semmes, K. N.; Hall, N. W.; Hickman, A. R.; Kaderli, M.; Windham, C. L.; Marunde, M. R.; Maryanski, D. N.; Noll, K.; Shannon, L.; Spengler, J.; Cowles, M. W.; Sun, Z.-W.; Keogh, M.-C.; Johnstone, A. L.; Weinzapfel, E. N.; Sun, L.
Show abstract
The precise regulation of chromatin composition is critical to gene expression and cellular identity, and thus a key component in development and disease. Robust assays to study chromatin features, including histone post-translational modifications (PTMs) and chromatin-associated proteins (e.g., transcription factors or PTM readers), are essential to understand their function and identify novel therapeutic strategies. To this end, Cleavage Under Targets and Release Using Nuclease (CUT&RUN) has emerged as a powerful tool for high-resolution epigenomic profiling. The approach has been successfully applied to numerous cell and tissue types, informing on target genomic distribution with unprecedented sensitivity and throughput. Here, we provide a detailed CUT&RUN protocol from sample collection through data analysis, including best practices and defined controls to ensure specific, efficient, and robust target profiling.
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