Abstract
The adult mammalian heart’s potential for regeneration is very inefficient. Importantly, adult mammalian cardiomyocytes (CMs) are characterized as a cell population with very limited mitotic potential. Conversely, the neonatal mouse heart possesses a brief, yet robust, regenerative capacity within the first week of life. Cell type-specific enrichment procedures are essential for characterizing the full spectrum of epigenomic landscapes and gene regulatory networks deployed by mammalian CMs. In this chapter, we describe a protocol useful for purifying CM nuclei from mammalian cardiac tissue. Furthermore, we detail a low-input procedure suitable for the parallel genome-wide profiling of chromatin accessibility, histone modifications, and transcription factor-binding sites. The CM nuclei purified using this process are suitable for multi-omic profiling approaches.
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Acknowledgment
We would like to thank Drs. Alisa Mo and Jeremy Nathans for providing the original nuclear isolation protocol. We thank Dr. Steven Henikoff for providing CUT&RUN reagents. This work was supported by grants from the National Institutes of Health (DE023177, HL127717, HL130804, HL118761 (J.F.M.)), F31HL136065 (M.C.H.), Vivian L. Smith Foundation (J.F.M.), State of Texas funding (J.F.M.), and Fondation LeDucq Transatlantic Networks of Excellence in Cardiovascular Research (14CVD01) “Defining the genomic topology of atrial fibrillation” (J.F.M.).
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Hill, M.C., Martin, J.F. (2021). Epigenetic Assays in Purified Cardiomyocyte Nuclei. In: Poss, K.D., Kühn, B. (eds) Cardiac Regeneration. Methods in Molecular Biology, vol 2158. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0668-1_23
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DOI: https://doi.org/10.1007/978-1-0716-0668-1_23
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0667-4
Online ISBN: 978-1-0716-0668-1
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