Nucleic Acids Research, 1994, Vol. 22, No. 15 2990-2997
© 1994
METHODS |
High sensitivity mapping of methylated cytosines
1Kanematsu Laboratories, Royal Prince Alfred Hospital Missenden Road, Camperdown, NSW 2050 2CSIRO Division of Biomolecular Engineering, Sydney Laboratory PO Box 184, North Ryde, NSW 2113 3School of Biological Sciences, A12, University of Sydney NSW 2006, Australia
*To whom correspondence should be addressed at: CSIRO Division of Biomolecular Engineering, Sydney Laboratory, PO Box 184, North Ryde, NSW 2113, Australia
Received May 16, 1994. Revised July 1, 1994. Accepted July 1, 1994.
An understanding of DNA methylation and its potential role in gene control during development, aging and cancer has been hampered by a lack of sensitive methods which can resolve exact methylation patterns from only small quantities of DNA. We have now developed a genomic sequencing technique which is capable of detecting every methylated cytosine on both strands of any target sequence, using DNA isolated from fewer than 100 cells. In this method, sodium bisulphite is used to convert cytosine residues to uracil residues in single-stranded DNA, under conditions whereby 5-methylcytosine remains non-reactive. The converted DNA is amplified with specific primers and sequenced. All the cytosine residues remaining in the sequence represent previously methylated cytosines in the genome. The work described has defined procedures that maximise the efficiency of denaturetion, bisulphite conversion and amplification, to permit methylation mapping of single genes from small amounts of genomic DNA, readily available from germ cells and early developmental stages.
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C Diatloff-Zito, A Nicole, G Marcelin, H Labit, E Marquis, C Bellanne-Chantelot, and J J Robert Genetic and epigenetic defects at the 6q24 imprinted locus in a cohort of 13 patients with transient neonatal diabetes: new hypothesis raised by the finding of a unique case with hemizygotic deletion in the critical region J. Med. Genet., January 1, 2007; 44(1): 31 - 37. [Abstract] [Full Text] [PDF]
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M. Oda, A. Yamagiwa, S. Yamamoto, T. Nakayama, A. Tsumura, H. Sasaki, K. Nakao, E. Li, and M. Okano DNA methylation regulates long-range gene silencing of an X-linked homeobox gene cluster in a lineage-specific manner Genes & Dev., December 15, 2006; 20(24): 3382 - 3394. [Abstract] [Full Text] [PDF]
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 R. Claus, M. Fliegauf, M. Stock, J. A. Duque, M. Kolanczyk, and M. Lubbert Inhibitors of DNA methylation and histone deacetylation independently relieve AML1/ETO-mediated lysozyme repression J. Leukoc. Biol., December 1, 2006; 80(6): 1462 - 1472. [Abstract] [Full Text] [PDF]
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B. Reinhart, A. Paoloni-Giacobino, and J. R. Chaillet Specific Differentially Methylated Domain Sequences Direct the Maintenance of Methylation at Imprinted Genes Mol. Cell. Biol., November 15, 2006; 26(22): 8347 - 8356. [Abstract] [Full Text] [PDF]
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V. W. Wen, K. Wu, S. Baksh, R. A. Hinshelwood, R. B. Lock, S. J. Clark, M. A.S. Moore, and K. L. MacKenzie Telomere-Driven Karyotypic Complexity Concurs with p16INK4a Inactivation in TP53-Competent Immortal Endothelial Cells. Cancer Res., November 15, 2006; 66(22): 10691 - 10700. [Abstract] [Full Text] [PDF]
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P. Novak, T. Jensen, M. M. Oshiro, R. J. Wozniak, M. Nouzova, G. S. Watts, W. T. Klimecki, C. Kim, and B. W. Futscher Epigenetic Inactivation of the HOXA Gene Cluster in Breast Cancer Cancer Res., November 15, 2006; 66(22): 10664 - 10670. [Abstract] [Full Text] [PDF]
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N. C. Whitelaw and E. Whitelaw How lifetimes shape epigenotype within and across generations Hum. Mol. Genet., October 15, 2006; 15(suppl_2): R131 - R137. [Abstract] [Full Text] [PDF]
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 Q. Fu, R. A. McKnight, X. Yu, C. W. Callaway, and R. H. Lane Growth retardation alters the epigenetic characteristics of hepatic dual specificity phosphatase 5 FASEB J, October 1, 2006; 20(12): 2127 - 2129. [Abstract] [Full Text] [PDF]
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M. Osanai, M. Murata, N. Nishikiori, H. Chiba, T. Kojima, and N. Sawada Epigenetic Silencing of Occludin Promotes Tumorigenic and Metastatic Properties of Cancer Cells via Modulations of Unique Sets of Apoptosis-Associated Genes. Cancer Res., September 15, 2006; 66(18): 9125 - 9133. [Abstract] [Full Text] [PDF]
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Y. Shen, J. Chow, Z. Wang, and G. Fan Abnormal CpG island methylation occurs during in vitro differentiation of human embryonic stem cells Hum. Mol. Genet., September 1, 2006; 15(17): 2623 - 2635. [Abstract] [Full Text] [PDF]
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D. M. Maatouk, L. D. Kellam, M. R. W. Mann, H. Lei, E. Li, M. S. Bartolomei, and J. L. Resnick DNA methylation is a primary mechanism for silencing postmigratory primordial germ cell genes in both germ cell and somatic cell lineages Development, September 1, 2006; 133(17): 3411 - 3418. [Abstract] [Full Text] [PDF]
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T. Rauch, H. Li, X. Wu, and G. P. Pfeifer MIRA-Assisted Microarray Analysis, a New Technology for the Determination of DNA Methylation Patterns, Identifies Frequent Methylation of Homeodomain-Containing Genes in Lung Cancer Cells Cancer Res., August 15, 2006; 66(16): 7939 - 7947. [Abstract] [Full Text] [PDF]
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N. Shimada, H. Matsudo, K. Osano, H. Arakawa, J.-M. Buerstedde, Y. Matsumoto, K. Chayahara, A. Torihata, and M. Ono Activation of the chicken Ig-{beta} locus by the collaboration of scattered regulatory regions through changes in chromatin structure Nucleic Acids Res., August 11, 2006; 34(13): 3794 - 3802. [Abstract] [Full Text] [PDF]
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J. A. Gollob, C. J. Sciambi, B. L. Peterson, T. Richmond, M. Thoreson, K. Moran, H. K. Dressman, J. Jelinek, and J.-P. J. Issa Phase I Trial of Sequential Low-Dose 5-Aza-2'-Deoxycytidine Plus High-Dose Intravenous Bolus Interleukin-2 in Patients with Melanoma or Renal Cell Carcinoma Clin. Cancer Res., August 1, 2006; 12(15): 4619 - 4627. [Abstract] [Full Text] [PDF]
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J. C. Brandes, J. G. Herman, D. R. Hodge, and W. L. Farrar p53 Expression after Treatment with Zebularine Is Not Due to Demethylation. Cancer Res., July 1, 2006; 66(13): 6892 - 6893. [Full Text] [PDF]
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 A. Tsumura, T. Hayakawa, Y. Kumaki, S.-i. Takebayashi, M. Sakaue, C. Matsuoka, K. Shimotohno, F. Ishikawa, E. Li, H. R. Ueda, et al. Maintenance of self-renewal ability of mouse embryonic stem cells in the absence of DNA methyltransferases Dnmt1, Dnmt3a and Dnmt3b. Genes Cells, July 1, 2006; 11(7): 805 - 814. [Abstract] [Full Text] [PDF]
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F. A. Champagne, I. C. G. Weaver, J. Diorio, S. Dymov, M. Szyf, and M. J. Meaney Maternal Care Associated with Methylation of the Estrogen Receptor-{alpha}1b Promoter and Estrogen Receptor-{alpha} Expression in the Medial Preoptic Area of Female Offspring Endocrinology, June 1, 2006; 147(6): 2909 - 2915. [Abstract] [Full Text] [PDF]
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J. Shu, J. Jelinek, H. Chang, L. Shen, T. Qin, W. Chung, Y. Oki, and J.-P. J. Issa Silencing of bidirectional promoters by DNA methylation in tumorigenesis. Cancer Res., May 15, 2006; 66(10): 5077 - 5084. [Abstract] [Full Text] [PDF]
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