International Mammalian Genome Society

17th International Mouse Genome Conference

9-12 November 2003, Braunschweig, Germany


POSTER 36 - IDENTIFICATION OF GENOMIC SITES OF TISSUE SPECIFIC DNA METHYLATION USING REAL AND VIRTUAL RESTRICTION LANDMARK GENOMIC SCANNING

Held W A
Roswell Park Cancer Insitute

Co-Authors: 1) Song F, 1) Kimura MT 1) Smith J 2) Matsuyama T, 1) Nagase H
Institutions: 1) Roswell Park Cancer Insitute, 2) RIKEN (The Institute of Physical and Chemical Research

Changes in DNA methylation during development suggest a role in development and tissue-specific differentiation. Restriction Landmark Genomic Scanning (RLGS) is a method for the 2-dimensional display of end-labelled DNA restriction fragments. When using a methylation sensitive restriction enzyme such as NotI as the restriction landmark, RLGS primarily displays CpG islands and associated regions and can be used to detect developmental and tissue-specific differences in DNA methylation. Real RLGS was performed using DNA from several different mouse tissues (C57Bl/6J) to identify RLGS loci with tissue specific differences in DNA methylation status. The genomic DNA sequences corresponding to these RLGS loci have been rapidly determined using new computational software that displays a virtual RLGS image of mouse genome sequence information. The DNA sequence of 40 tissue-specific RLGS loci were confirmed by PCR amplification of eluted Spot DNA using primers determined from the “Virtual sequence”. The vast majority of the loci identified thus far are located within CpG islands (90%) associated with the 5' promoter region (70%) or 3' terminus (25%) of genes. Bisulfite sequencing was used to confirm the differential tissue-specific methylation status of some of the loci and to evaluate the density and extent of the methylation in the region. For some loci, the methylation status of the 5' promoter region inversely correlates with the tissue expression profile of the gene determined by publicly available microarray expression profiles. However, it is likely that developmental and tissue-specific differences in DNA methylation have additional roles in chromatin structure and epigenetic programming.


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