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成果報告書詳細
管理番号20170000000728
タイトル*平成28年度中間年報 植物等の生物を用いた高機能品生産技術の開発 植物の生産性制御に係る共通基盤技術開発 遺伝子発現制御および栽培環境制御の融合による代謝化合物高生産基盤技術開発
公開日2017/8/11
報告書年度2016 - 2016
委託先名国立大学法人北海道大学
プロジェクト番号P16009
部署名材料・ナノテクノロジー部
和文要約
英文要約Title:Development of the techniques to edit DNA demethylation status at specific sites

Methylation of DNA or histone proteins in the region such as promoters often causes transcriptional gene silencing (TGS), the epigenetic regulation system of gene expression. Because epigenetic regulation was involved in the control of many genes related to synthesis and accumulation of useful compounds in plants, development of the editing techniques of DNA methylation pattern is needed to improve useful plants. DNA methylation is induced through the RNA directed DNA methylation (RdDM) pathway, and scaffold RNA plays an important role to direct the region targeted by RdDM. In order to edit the status of DNA methylation site-specifically, we aim to develop the techniques disturbing the function of specific scaffold RNAs by a ribozyme and/or dummy RNA. Because the model systems for DNA methylation analysis are necessary to evaluate the efficiency of those techniques, two model systems were constructed using Arabidopsis and Nicotiana benthamiana in this study.
In Arabidopsis, five transgenic plants overexpressing the green fluorescence protein (GFP) gene driven by the 35S promoter were developed, and the plant showing GFP TGS was selected as a model experimental line. In N. benthamiana, we constructed the system inducing TGS of 35S-GFP using the Cucumber mosaic virus (CMV) vector. The CMV vectors carrying various lengths of the 35S segment were infected onto N. benthamiana line 16c expressing 35S-GFP and the GFP silencing plants were selected. Because the status of TGS was stably inherited to the next generation, we decided to use these lines as models in our study. Using one of these GFP silencing lines, the accumulation pattern of siRNA within the 35S promoter region was investigated. siRNA was mostly produced from the 5’ end side of the 35S promoter and there was also a small peak of siRNA at the 3’ end side. Because it was possible that scaffold RNA were generated from those sites with high frequency, those scaffold RNAs should be targeted by a ribozyme and/or dummy RNA.
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