Analysis of insulator function of non-coding DNA using genome editing

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K06602
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43050:Genome biology-related
• Research Institution: Hiroshima University
• Principal Investigator: Naoaki Sakamoto 広島大学, 統合生命科学研究科(理), 准教授 (00332338)
• Co-Investigator(Kenkyū-buntansha): 粟津 暁紀 広島大学, 統合生命科学研究科(理), 准教授 (00448234)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: インスレーター

Outline of Final Research Achievements

The purpose of this study was to clarify the physical properties and mechanism of action of the insulator. We attempted to apply CRISPR-Cas3, which can introduce a wide range of large deletion mutations, to sea urchins, but it was not successful, indicating that further improvement is needed. Furthermore, we tried to use knock-in for functional deletion, but although knock-in products were detected, their efficiency was very low. Various treatments to inhibit NHEJ were carried out, but no significant improvement in efficiency was obtained.
To correlate physical properties of DNA with their insulator activity, we used lambda DNAs exhibiting various physical properties, and we found that each of them had a different effect on enhancer/promoter communication.

Free Research Field

分子生物学

Academic Significance and Societal Importance of the Research Achievements

ウニは、ヒトを含む新口動物の祖先型の動物であり、その進化的位置付けからもゲノムの進化を考える上で重要な動物である。ゲノム編集の活用により、内在の染色体環境下で遺伝子の発現制御機構について解析できるシステムが構築できれば、より正確なゲノム機能を解析でき、膨大な非コードDNAの役割とその進化を明らかにできると考えられる。また、ウニにおけるゲノム編集技術の適用法がさらに広まるれば、有用品種の作出など産業面でも価値のあることであると考えられる。