DNA Methylation Profiling Method
DNA Methylation Profiling Method
The invention comprises new methods for profiling whole genome methylation by converting unmethylated cytosines in CpG dinucleotides to thymine analogues. Previous studies have shown that DNA methyltransferases can efficiently transfer a wide variety of functional groups to the 5 position of cytosines in DNA by means of synthetic S-Adenosyl L-methionine (AdoMet) analogs in which the methyl group has been replaced.
New York, NY, United States
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Background
Whole Genome Methylation Profiling In the effort to gain insight into whole genome methylation patterns, their biological functions and their abnormality in human disease, various methylation profiling technologies have been developed. However, existing methods have either limitations or severe drawbacks. Traditional bisulfite conversion suffers from variability due to incomplete conversion, over-conversion, DNA degradation, and low yield. The ideal whole genome methylation profiling method would combine ultrahigh throughput sequencing, and site specificity to give unparalleled information concerning methylation patterns. 

Conversion of Unmethylated Cytosines in CpG Dinucleotides to Thymine Analogues The invention comprises new methods for profiling whole genome methylation by converting unmethylated cytosines in CpG dinucleotides to thymine analogues. Previous studies have shown that DNA methyltransferases can efficiently transfer a wide variety of functional groups to the 5 position of cytosines in DNA by means of synthetic S-Adenosyl L-methionine (AdoMet) analogs in which the methyl group has been replaced. By using such AdoMet derivatives, the CpG-specific DNA methyltransferase SssI can deliver the chemically reactive group to cytosine residues, which then undergo conversion to a thymine analog or to a new fifth base (neobase X). 

Applications
• Profile whole genome methylation patterns 
• Determine aberrant methylation patterns in disease states 
• Diagnostic platform technology 

Advantages: 
• More accurate conversion compared to traditional bisulfite conversion 
• Only unmethylated CpG sites will be converted instead of all CpG sites 
• More information content in short sequence tags; easier tag-mapping 
• Less toxic and less DNA damage than in traditional bisulfite conversion 
• Enables sequencing of longer DNA fragments
 
Lead InventorsTimothy Bestor, Ph.D.Jingyue Ju, Ph.D.

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