Technology Commercialization and Partnerships | BSA 12-38: Method for Methyl-CpG DNA Enrichment

BSA 12-38: Method for Methyl-CpG DNA Enrichment

BNL Reference Number: BSA 12-38

Patent Status: Application Number 20130040343 was published on February 14, 2013

Summary

This invention provides methods and means to collect and concentrate the DNA sequences containing Methyl-CpG regions from a sample of DNA. The methylation of specific CpG sequences of genomic DNA has been correlated with various disease states, essentially representing biomarkers for diseases. As the number of confirmed such correlations increases, so will demand for implementing screening assay panels that assess the methylation status of all established biomarker sequences. This invention affords the methods to quickly collect the population of sequences to be subjected to the screening assays. The method is simple and could be automated.

Description

his invention is a member of the family of inventions relating to detection of Methyl-CpG dinucleotides in DNA. As a divisional invention from the parent invention (BSA 07-32-Methyl-CpG detection using McrA and BSA 11-19-Methyl-CpG detection using McrA) this invention uses well-known techniques to concentrate the portion of a DNA sample that contains the methylated dinucleotide regions. By various means to re-methylate cytosine residues following bisulfite treatment, specific arrangements or contexts of the methyl-CpG sequences can be assessed either individually or as a group. Using next generation sequencing methods, one can rapidly assess a panel of recognized methyl-CpG biomarker regions. After fragmenting a DNA sample, the DNA is bisulfite treated to convert all cytosine residues to uracil. After amplification of the DNA, using linkers to amplify all DNA or amplification primers to amplify only select sequences, as desired, all remaining cytosine residues were those that were methylated in the original sample of DNA. Various enzymatic or other methods may then be used to methylate the cytosine residues that are present as CpG dinucleotides. The methylated sequences can be separated from the unmethylated sequences using specific binding partners. Using specific methyl-CpG binding assays (such as those afforded by use of McrA) or standard sequencing methods, the methylation status of the original sample sequences can be determined.

Benefits

Coupled with the two other inventions in this patent suite, this invention provides methods that enhance the utility of the patents covering the specific Methyl-CpG McrA binding protein. The importance of the assays and screens for identification of significant regions of methylation of CpG dinucleotides will increase substantially as more biomarker methyl-CpG sequences are confirmed.

Applications and Industries

Companies focusing on development of biomarker assays will benefit, particularly those interested in developing screening assays for cancer-related markers.