Published online 20 January 2006
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Microarray-based DNA methylation profiling: technology and applications
The Krembil Family Epigenetics Laboratory, Centre for Addiction and Mental Health 250 College Street, Toronto, ON, Canada M5T 1R8 1Affymetrix Santa Clara, USA 2The Microarray Centre, The University Health Network 200 Elizabeth Street, Toronto, ON, Canada M5G 2C4
*To whom correspondence should be addressed. The Krembil Family Epigenetics Laboratory, Room 28, Centre for Addiction and Mental Health, 250 College Street, Toronto, ON, Canada M4T 1R8. Tel: +1 416 5358501 4880; Fax: +1 416 979 4666; Email: Arturas_Petronis{at}camh.net
Received November 21, 2005. Revised December 20, 2005. Accepted January 5, 2006.
This work is dedicated to the development of a technology for unbiased, high-throughput DNA methylation profiling of large genomic regions. In this method, unmethylated and methylated DNA fractions are enriched using a series of treatments with methylation sensitive restriction enzymes, and interrogated on microarrays. We have investigated various aspects of the technology including its replicability, informativeness, sensitivity and optimal PCR conditions using microarrays containing oligonucleotides representing 100 kb of genomic DNA derived from the chromosome 22 COMT region in addition to 12 192 element CpG island microarrays. Several new aspects of methylation profiling are provided, including the parallel identification of confounding effects of DNA sequence variation, the description of the principles of microarray design for epigenomic studies and the optimal choice of methylation sensitive restriction enzymes. We also demonstrate the advantages of using the unmethylated DNA fraction versus the methylated one, which substantially improve the chances of detecting DNA methylation differences. We applied this methodology for fine-mapping of methylation patterns of chromosomes 21 and 22 in eight individuals using tiling microarrays consisting of over 340 000 oligonucleotide probe pairs. The principles developed in this work will help to make epigenetic profiling of the entire human genome a routine procedure.
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