TY  - JOUR
A1  - Shiratori, Hiromi
A1  - Feinweber, Carmen
A1  - Knothe, Claudia
A1  - Lötsch, Jörn
A1  - Thomas, Dominique Jeanette
A1  - Geisslinger, Gerd
A1  - Parnham, Michael J.
A1  - Resch, Eduard
T1  - High-throughput analysis of global dna methylation using methyl-sensitive digestion
T2  - PLoS one
N2  - DNA methylation is a major regulatory process of gene transcription, and aberrant DNA methylation is associated with various diseases including cancer. Many compounds have been reported to modify DNA methylation states. Despite increasing interest in the clinical application of drugs with epigenetic effects, and the use of diagnostic markers for genome-wide hypomethylation in cancer, large-scale screening systems to measure the effects of drugs on DNA methylation are limited. In this study, we improved the previously established fluorescence polarization-based global DNA methylation assay so that it is more suitable for application to human genomic DNA. Our methyl-sensitive fluorescence polarization (MSFP) assay was highly repeatable (inter-assay coefficient of variation = 1.5%) and accurate (r2 = 0.99). According to signal linearity, only 50–80 ng human genomic DNA per reaction was necessary for the 384-well format. MSFP is a simple, rapid approach as all biochemical reactions and final detection can be performed in one well in a 384-well plate without purification steps in less than 3.5 hours. Furthermore, we demonstrated a significant correlation between MSFP and the LINE-1 pyrosequencing assay, a widely used global DNA methylation assay. MSFP can be applied for the pre-screening of compounds that influence global DNA methylation states and also for the diagnosis of certain types of cancer.
Y1  - 2016
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/41720
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-417208
SN  - 1932-6203
N1  - Copyright: © 2016 Shiratori et al. This is an open access article distributed under the terms of the Creative Commons Attribution License  http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
VL  - 11
IS  - (10): e0163184
SP  - 1
EP  - 16
PB  - PLoS
CY  - Lawrence, Kan.
ER  -