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Using Immunofluorescence to Observe Methylation Changes in Mammalian Preimplantation Embryos

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Book cover Nuclear Reprogramming

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 325))

Abstract

DNA methylation is an important epigenetic mark involved in gene silencing, X chromosome and transposon inactivation, genomic imprinting, and chromosome stability. Recently, it has been increasingly recognized that DNA methylation plays an essential regulatory function in mammalian development and cancer biology, serving to repress nontranscribed genes stably in differentiated adult somatic cells. DNA methylation is subject to reprogramming during development, involving both demethylation (active and passive) and de novo methylation phases. Recent data show deregulation of genome-wide DNA methylation levels in embryos generated by somatic nuclear transfer, suggesting this epigenetic mark and its regulation are essential to ensure viable embryos. Immunofluorescence using antibodies against 5-methylcytosine is an invaluable method for the large-scale screening of genome-wide methylation. This method has a high degree of reproducibility, essential for the analysis of small numbers of valuable samples, providing information on methylation profiles of individual cells and embryos.

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© 2006 Humana Press Inc.

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Santos, F., Dean, W. (2006). Using Immunofluorescence to Observe Methylation Changes in Mammalian Preimplantation Embryos. In: Pells, S. (eds) Nuclear Reprogramming. Methods in Molecular Biology™, vol 325. Humana Press. https://doi.org/10.1385/1-59745-005-7:129

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  • DOI: https://doi.org/10.1385/1-59745-005-7:129

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-379-4

  • Online ISBN: 978-1-59745-005-8

  • eBook Packages: Springer Protocols

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