Exploring the Elements Required for Accurate Genome Editing Using Autonomous Activated CRISPR

2022 Fiscal Year Final Research Report

• Project/Area Number: 21K14740
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: Hiroshima University
• Principal Investigator: Daisuke Matsumoto 広島大学, 医系科学研究科(薬), 助教 (20889381)
• Project Period (FY): 2021-04-01 – 2023-03-31
• Keywords: ゲノム編集 / CRISPR-Cas9 / Anti-CRISPR / 相同性組換え / 細胞周期

Outline of Final Research Achievements

We evaluated whether a synergistic effect on homologous recombination efficiency could be obtained by combining the cell cycle-dependent activation of CRISPR using Anti-CRISPR with the cell cycle-dependent regulation of Cas9 expression itself. The results showed that cell cycle-dependent expression of Cas9 itself improved homologous recombination efficiency compared to normal Cas9, but off-target effects occurred at a higher frequency. On the other hand, the combination with the activation system using Anti-CRISPR further improved the homologous recombination efficiency and reduced the off-target effects.

Free Research Field

遺伝子工学

Academic Significance and Societal Importance of the Research Achievements

ゲノム編集を臨床といった正確な遺伝子の書き換えを行う上で、相同性組換えを介した修復は重要である。本研究成果によって、細胞周期を厳密に限定したCRISPR-Cas9の活性制御によって相同性組換えの向上とオフターゲット作用の低減が可能であることが示唆された。この結果を踏まえ、より詳細な遺伝子修復とCRISPRを用いたDNA切断の関係性を解明していくことによって、より安全なゲノム編集技術の開発に貢献できると期待される。