Imaging analysis for nuclear dynamics in structural change of nuclei in early embryo and differentiation.

2019 Fiscal Year Final Research Report

• Project/Area Number: 17K07241
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Genome biology
• Research Institution: Hiroshima University
• Principal Investigator: Sakamoto Naoaki 広島大学, 統合生命科学研究科(理), 准教授 (00332338)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Keywords: ウニ / 核内構造 / ゲノム編集 / CRISPR-Cas9

Outline of Final Research Achievements

We found that early histone genes are transcriptionally active during morula stage and that multiple early histone gene loci interact each other during this stage. Furthermore, transcriptional active state of these gene loci is involved in this chromosomal interactions. For live imaging analysis of these loci, we attempted to establish the CRISPR-Cas9 system and achieved 100% knockout efficiency in sea urchin embryos using CRISPR-Cas9. Currently, we constructed a imaging tool based on this Cas9 that is available for sea urchin and the tuning of this tool is still ongoing. In addition, we analyzed the intranuclear structure of sea urchin embryos by using fluorescent proteins and found that the centromere showed a biased distribution in the nucleus like Rabl configuration and that multiple spots of the nucleolus tended to be close together.

Free Research Field

分子生物学

Academic Significance and Societal Importance of the Research Achievements

遺伝子を取り巻く核内環境の変化が劇的に変化するウニ初期胚において、遺伝子の活性状態と染色体間相互作用が関連しているのは、非常に興味深いと思われる。また、CRISPR-Cas9システムによる効率的ゲノム編集法がバフンウニで確立できたことは、各種海産動物の育種においてもこのCRISPR-Cas9システムが応用可能であることを示唆しており、興味深い。