Abstract
R-loops are three-stranded nucleic acid structures that consist of a DNA–RNA hybrid and a displaced single-stranded DNA. R-loops occur during transcription and participate in multiple physiological processes such as DNA repair, modulating DNA topology, and regulation of gene transcription. Dysfunctional R-loops associate with several human diseases such as neurological disorders and cancer. Therefore, accurately and comprehensively profiling native R-loops is crucial to understand their functions under both physiological and pathological conditions. Here, we describe a convenient native R-loop profiling method, R-loop CUT&Tag, which combines a DNA–RNA hybrid sensor (GST-His6–2 × HBD or S9.6 antibody) with a pA-Tn5-based cleavage under targets and tagmentation approach. R-loop CUT&Tag starts with 0.5 million cells and can sensitively detect native and specific R-loops at the promoter, gene body, and enhancer regions.
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Acknowledgments
This work was funded by the National Key Research and Development Program of China Stem Cell and Translational Research (NO. 2019YFA0111100), NSFC Excellent Young Scientists Fund (82122006), and the China National Natural Science Foundation (82172641) awarded to K.L.
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Wang, H., Li, C., Liang, K. (2022). Genome-Wide Native R-Loop Profiling by R-Loop Cleavage Under Targets and Tagmentation (R-Loop CUT&Tag). In: Aguilera, A., Ruzov, A. (eds) R-Loops . Methods in Molecular Biology, vol 2528. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2477-7_23
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DOI: https://doi.org/10.1007/978-1-0716-2477-7_23
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