| 研究実績の概要 |
At this stage of the project, using CAGE data. we were able to show that, heat shock transcription factor (HSTF) is one of the key regulators of transcriptional activation in response to dehydration in the cells of the anhydrobiotic chironomid.We also confirmed it by RNAi experiment. Mapping the CAGE data updated genome assembly revealed new patterns of transcriptional activity in the tissues of chironomids during anhydrobiosis cycle. Most of the "anhydrobiosis-associated" were activated in dry tissues (75%), with ~ 39% of them were active in various tissues in a similar manner. We found some characteristics associated with tissue-specific activation of this group of genes: globins for for fat body, specific oxidoreductases, methyltransferase, LEA, in the brains.
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| 今後の研究の推進方策 |
The final year of the current project will be devoted to functional analysis of the identified regulators of anhydrobiosis. We are going to apply Total-Seq and small RNA analysis of Pv11 cells to identify a pool of non-coding RNA interacting with key anhydrobiotic genes. This year will will also implement protocol of CRISP-Cas9 knock out at least on of the key anhydrobiosis-involved transcription factor gene followed by CAGE analysis to identify the regulatory network associate with anhydrobiosis. Finally, we are going to proceed with phospho-proteomic analysis of Pv11 cells upon desiccation, to link gene expression data and actual protein abundance and proteins phosphorylation patter in the dry cells.
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