Development of innovative knock-in technology using endogenous homologous recombination mechanism

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K21370
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 42:Veterinary medical science, animal science, and related fields
• Research Institution: Shiga University of Medical Science
• Principal Investigator: TSUKIYAMA Tomoyuki 滋賀医科大学, 動物生命科学研究センター, 特任准教授 (60612132)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Keywords: ゲノム編集

Outline of Final Research Achievements

Although CRISPR/Cas9 and other genome editing technologies have enabled the generation of a variety of genetically modified animals, knock-in of large fragments by homologous recombination is still inefficient. Here, we focused on the endogenous homologous recombination mechanism and applied it to knock-in to establish a highly efficient gene modification technique.
In this study, we first constructed an evaluation system to efficiently assess knock-in efficiency. Additionally, we cloned several genes known to work in the endogenous homologous recombination mechanism and factors that work synergistically to induce a state close to endogenous homologous recombination.

Free Research Field

発生工学

Academic Significance and Societal Importance of the Research Achievements

申請者が所属する滋賀医科大学では、カニクイザルにおけるゲノム編集研究を積極的に推進しているが、サルを用いる場合、マウスのように単純に試行数を増やすことで効率の問題を回避するということが倫理的に難しく、現状の効率ではノックイン実験は難しい状況である。
もし効率よくノックインを誘導することが可能になれば、カニクイザルを含めた大動物のみならず、全てを置き換えるポテンシャルを秘めた意欲的な研究である。