Abstract
Cleavage under targets and release using nuclease (CUT&RUN) allows the chromatin profiling of proteins of interest for which specific antibodies are available. Because it is performed on intact chromatin in situ, CUT&RUN provides exceptional signal over background, making it an ideal choice for chromatin profiling on primary cells available at limited numbers. Here, we describe its application to the profiling of histone post-translational modifications in germ cells isolated from mouse embryos from 12.5 up to 18.5 days postfertilization. This approach can be applied to as low as 100 isolated germ cells, allowing the generation of multiple genome-wide profiles from the cells obtained from a single embryo.
Key words
- CUT&RUN
- Histone modifications
- Embryonic germ cells
- Chromatin profiling
- Low input
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Acknowledgments
We would like to thank François Dossin (Heard lab, EMBL Heidelberg) for advice on CUT&RUN modifications, Martin Möckel (Protein Expression Facility, IMB Mainz) for advice and technical help, and Violeta Morin for comments on the protocol. This work was supported by the Genomics, FACS, and Animal Core Facilities of the Institute of Molecular Biology in Mainz.
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Prakash, S.A., Barau, J. (2021). Chromatin Profiling in Mouse Embryonic Germ Cells by CUT&RUN. In: Ancelin, K., Borensztein, M. (eds) Epigenetic Reprogramming During Mouse Embryogenesis. Methods in Molecular Biology, vol 2214. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0958-3_17
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DOI: https://doi.org/10.1007/978-1-0716-0958-3_17
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