| 研究課題/領域番号 |
18K06188
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| 研究機関 | 国立研究開発法人理化学研究所 |
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研究代表者 |
SHARIF JAFAR 国立研究開発法人理化学研究所, 生命医科学研究センター, 専任研究員 (00577968)
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| 研究期間 (年度) |
2018-04-01 – 2021-03-31
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| キーワード | Epidermis / Basal layer / Differentiated cells / DNA methylation / H3K27me3 |
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| 研究実績の概要 |
The goal of the present research is two-fold. First, to elucidate the epigenetic mechanism that regulates barrier function in the epidermis. Second, to elucidate the role of genome organization in 3D for regulation of the barrier function. To elucidate the first goal, I have performed global DNA methylation analysis by RRBS-seq (reduced-representation bisulfite sequencing), as well as analysis of histone methylation (H3K27me3) by ChIP-seq.
I purified basal layer keratinocytes, by treating the epidermis with dispase I protease; and the differentiated cell layer by treating with ETA (exfoliative toxin A). RRBS-seq revealed a group of loci that gain or lose DNA methylation in differentiated cells, compared to basal layer cells. This indicates that global DNA methylation profile dynamically changes in association with keratinocyte differentiation.
ChIP-seq showed that upon differentiation, H3K27me3 levels were reduced from target gene promoters. H3K27me3 is involved in suppression of genes that are associated with differentiation, and therefore my findings are consistent with the previous findings on the function of H3K27me3.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
As mentioned above, I have two main goals in this project. The first goal, namely, epigenetic analysis of the basal layer cells and differentiated cells, is mostly completed. Therefore, the experimental progress is going on smoothly.
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| 今後の研究の推進方策 |
Having mostly completed the first goal of my project. I will move on to the second goal, namely, 3D nuclear analysis of the basal layer cells, and the differentiated cells. To this end, I will take advantage of Hi-C, followed by in house bioinformatics analysis, to reveal how the 3D organization of the genome changes in association to cellular differentiation.
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