Abstract
Genomic imprinting is an epigenetically controlled form of gene regulation leading to the preferential expression of one parental gene copy. To date, approximately 40 imprinted genes have been described that are exclusively or predominantly expressed from either the paternal or the maternal allele (www.mgu.har.mrc.ac.uk/imprinting/implink.html). Changes in the imprinted expression of such genes result in developmental abnormalities; in the human they are associated with several diseases and various types of cancer(1-3).
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Constancia, M., Pickard, B., Kelsey, G., and Reik, W. (1998) Imprinting mechanisms. Genome Res. 8, 881–900
Nicholls, R. D., Saitoh, S., and Horsthemke, B. (1998) Imprinting in Prader-Willi and Angelman syndromes. TIGS 14, 194–200.
Tycko, B. (1997) DNA methylation in genomic imprinting. Mutat. Res.-Rev. in Mutat. Res. 386, 131–140.
Reik, W.and Walter, J. (2001) Genomic imprinting; parental influence on the genome. Nature Rev. Genet. 2, 21–32.
Li, E. B. C.and Jaenisch, R. (1992) Targeted mutation of the DNA methyltransferase gene results in embryonic lethality. Cell 69, 915–926.
Frommer, M., McDonald, L. E., Millar, D. S., Collis, C. M., Watt, F., Grigg, G.W., Molloy, P. L., and Paul, C. L. (1992) A genomic sequencing protocol that yields a positive display of 5-methylcytosine residues in individual DNA strands. Proc. Natl. Acad. Sci.USA 89, 1827–1831.
Paul, C. L.and Clark, S. J. (1996) Cytosine methylation: Quantitation by automated genomic sequencing and GENESCAN analysis. BioTechniques 21, 126–133.
Gonzalgo, M.and Jones, P. A. (1997) Rapid quantitation of methylation differences at specific sites using methylation-sensitive single nucleotide primer extension (Ms-SNuPE) Nucleic Acids Res. 25, 2529–2531.
Clark, S. J., Harrison, J., Paul, C. L., and Frommer, M. (1994) High sensitivity mapping of methylated cytosines. Nucl. Acids Res. 22, 2990–2997
Feil, R., Walter, J., Allen, 6N. D., and Kelsey, G. (1994) Developmental control of allelic methylation in the imprinted mouse Igf2 and H19 genes. Development 120, 2933–2943
Raizis, A. M., Schmitt, F., and Jost, J. P. (1994) A bisulfite method of 5-methylcytosine mapping that minimises template degradation. Anal. Biochem. 226, 161–166.
Olek, A. Oswald, J., and Walter, J. (1996) A modified and improved method for bisulfite based cytosine methylation analysis. Nucleic Acids Res. 24, 5064–5066.
Paulin, R., Grigg, G.W., Davey, M. W., and Piper, A. A. (1998) Urea improves efficiency of bisulfite-mediated sequencing of 5-methylcytosine in genomic DNA. Nucl. Acids Res. 26, 5009–5010.
Hayatsu, H., Wataya, Y., Kai, K., and Iida, S. (1970) Reaction of sodium bisulfite with uracil, cytosine, and their derivatives. Biochemistry 9, 2858–2864.
Wang, R. Y.-H., Gehrke, C. W., and Ehrlich, M. (1980) Comparison of bisulfite modification of 5-methyldeoxycytidine and deoxycytidine residues. Nucleic Acids Res. 8, 4777–4790.
Sambrook, G., Fritsch, E. F., and Maniatis, T. (1988) Molecular Cloning and Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Warnecke, P. M., Stirzaker, C., Melki, J. R., Millar, D. S., Paul, C. L., and Clark, S. J.(1997) Detection and measurement of PCR bias in quantitative methylation analysis of bisulfite-treated DNA. Nucl. Acids Res. 25, 4422–4426.
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© 2002 Humana Press Inc., Totowa, NJ
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Engemann, S., El-Maarri, O., Hajkova, P., Oswald, J., Walter, J. (2002). Bisulfite-Based Methylation Analysis of Imprinted Genes. In: Ward, A. (eds) Genomic Imprinting. Methods in Molecular Biology™, vol 181. Humana Press. https://doi.org/10.1385/1-59259-211-2:217
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DOI: https://doi.org/10.1385/1-59259-211-2:217
Publisher Name: Humana Press
Print ISBN: 978-0-89603-741-0
Online ISBN: 978-1-59259-211-1
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