Assessing the DNA methylation status of single cells with the comet assay |
| |
Authors: | Johannes F Wentzel Chrisna Gouws Etresia van Dyk Pieter J Pretorius |
| |
Institution: | a Division for Biochemistry, School of Physical and Chemical Sciences, North-West University, Potchefstroom 2520, South Africa b Statistical Consultation Services, North-West University, Potchefstroom 2520, South Africa |
| |
Abstract: | The comet assay (single cell gel electrophoresis) is a cost-effective, sensitive, and simple technique that is traditionally used for analyzing and quantifying DNA damage in individual cells. The aim of this study was to determine whether the comet assay could be modified to detect changes in the levels of DNA methylation in single cells. We used the difference in methylation sensitivity of the isoschizomeric restriction endonucleases HpaII and MspI to demonstrate the feasibility of the comet assay to measure the global DNA methylation level of individual cells. The results were verified with the well-established cytosine extension assay. We were able to show variations in DNA methylation after treatment of cultured cells with 5-azacytidine and succinylacetone, an accumulating metabolite in human tyrosinemia type I. |
| |
Keywords: | Comet assay DNA methylation HepG2 Cytosine extension assay Tyrosinemia type I Succinylacetone |
本文献已被 ScienceDirect 等数据库收录! |
|