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A sensitive microfluidic platform for a high throughput DNA methylation assay.

Maria Ronen1, Dorit Avrahami, Doron Gerber

  • 1The Mina & Everard Goodman Faculty of Life Sciences, The Nanotechnology Institute, Bar-Ilan University, Ramat Gan, 5290002, Israel. Doron.Gerber@biu.ac.il.

Lab on a Chip
|May 21, 2014
PubMed
Summary

We developed a microfluidic assay for DNA methylation studies. This high-throughput platform efficiently screens for epigenetic drug targets and characterizes enzyme kinetics, aiding cancer research.

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Area of Science:

  • Epigenetics
  • Molecular Biology
  • Biochemistry

Background:

  • DNA methylation is crucial for development and disease, like cancer.
  • Novel therapeutic targets require efficient screening platforms for DNA methylation modulators.

Purpose of the Study:

  • To present a universal, high-throughput, microfluidic-based fluorometric assay for DNA methylation research.
  • To demonstrate its utility for enzymatic activity characterization and inhibitor screening.

Main Methods:

  • Development of a microfluidic assay for DNA methylation.
  • Enzymatic activity measurement of HPAII DNA methyltransferase.
  • High-throughput screening of small molecule inhibitors using a two-step approach.

Main Results:

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  • The assay accurately measured HPAII DNA methyltransferase kinetics (K(m)(DNA) = 5.8 nM, K(m)(SAM) = 9.8 nM, Kcat = 0.04 s(-1)).
  • Identified and characterized non-nucleoside inhibitors SGI-1027 (IC50 = 4.5 μM) and RG108 (IC50 = 87.5 nM).

Conclusions:

  • The microfluidic assay is a versatile tool for quantitative enzymatic characterization.
  • It enables high-throughput screening and characterization of DNA methylation inhibitors for epigenetic studies.