예쁜꼬마선충을 통한 DNMT3a의 이소성 발현과 기능 분석

Abstract

DNA methylation is a key mechanism of epigenetics to regulate gene function and a major carrier of epigenetic information. The past studies shed light on function of DNMTs using Drosophila, mutated mice or cell lines. However, we still do not understand the mechanism of DNA methylation and gene expression modulated by DNMTs. Thus, transgenic C.elegans expressing mouse DNMT3a was generated to investigate DNA methylation mechanism and functions of DNMTs. In this study, activity of DNMT3a in transgenic C.elegans containing mDNMT3a was confirmed to initiate functional study of DNMTs through several experiments. Firstly, full length mDNMT3a gene in the worm was identified. And, by blotting and DNMT assays, I determined DNMT3a was actively expressed in nuclei. This work was also showed DNMT3a could methylate non-CpG sites. Furthermore, DNMT assay revealed DNMT activity would be in not only transgenic worm but also wild-type worm, even though it need further study to verify. Finally, I tried to find methylcytosine. NICEA result was indicated a possibility that methylcytosine was in transgenic C.elegans. However, I could not detect significant result about how many methylcytosine exist in transgenic C.elegans. In conclusion, the present study is a start point of functional study to investigate whether transgenic C.elegans is affected by DNMTs because mDNMT3a expression and activity in transgenic C.elegans were determined.