| 研究課題/領域番号 |
15K08535
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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 |
SEO WOOSEOK 国立研究開発法人理化学研究所, 統合生命医科学研究センター, 研究員 (40574116)
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| 研究期間 (年度) |
2015-10-21 – 2018-03-31
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| キーワード | アレルギー / 免疫関連疾患 |
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| 研究実績の概要 |
The main goal of this project is to study whether Runx transcription factor regulates CCL5 expression since Runx-deficient mouse shows dysregulated CCL5 expression and severe inflammatory diseases. In the previous year, I have generated a mouse line in which Runx-binding site of CCL5 gene is removed and confirmed that the removed Runx-binding site has critical roles in the regulation of CCL5 expression. The first goal of this fiscal year (H28) was to generate and analyze a BAC (Bacterial Artificial Chromosome) transgenic mouse line in which CCL5 gene is replaced by GFP-reporter and Runx-binding region on CCL5 locus is mutated at the specific sequence to specifically abrogate the Runx-binding without removing a piece of the genome. The mouse line was successfully generated and being expanded for detailed analysis. The second objective of this fiscal year was to perform ChIP (chromatic immunoprecipitation)-seq to confirm the direct binding of Runx transcription factor on CCL5 locus. ChIP-seq was successfully performed as planned and the results clearly show the direct binding of Runx transcription factor on CCL5 locus in vivo as expected. These two results clearly indicate that the binding of Runx plays a major role in the regulation of CCL5 expression.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
1: 当初の計画以上に進展している
理由
The objective of the first fiscal year (H27) was successfully completed by the generation and analysis of the knockout mouse for Runx-binding region of the endogenous CCL5. This study was successfully duplicated in BAC transgenic reporter system I generated in this second fiscal year (H28), further strengthening our model. With the two mice lines generated in H27 and H28, we have a clear conclusion that Runx transcription factor plays a major role in CCL5 expression. Furthermore, ChIP-seq performed this year directly proves that Runx binds to CCL5 locus in vivo. ChIP-seq evidence together with two genetically modified mouse lines, we can claim that inflammatory pathologies observed in Runx-deficient mice is due to the abrogation of Runx-mediated regulation of CCL5 expression.
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| 今後の研究の推進方策 |
Even though I have to do more detailed analysis on the newly generated BAC transgenic mouse line, I believe that we now have a clear conclusion that Runx-mediated transcriptional regulation of CCL5 is important to maintain proper levels of CCL5. This fiscal year (H29), we would like to further test this conclusion by performing an infection model. We plan to collaborate with Dr. Kubo’s laboratory in RIKEN-IMS to perform influenza infection model. We expect that the dysregulated CCL5 expression in Runx-deficient mice or new mice lines we generated would have effects on the immune responses toward influenza viruses. Examination of our model in a real infection model would reinforce our claim and highlight the importance of our study.
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| 次年度使用額が生じた理由 |
ChIP (chromatin immnoprecipitation) - seq experiments were planned to perform more than three times in the second fiscal year, but it was performed only once so far due to technical reasons.
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| 次年度使用額の使用計画 |
I will perform two or more runs of ChIP-seq experiments by using IMS central facility and this incurred amount of the grant will be used to pay for the services.
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