In vivo genomic engineering with a novel genome editing tools

2021 Fiscal Year Final Research Report

• Project/Area Number: 19K16025
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 42040:Laboratory animal science-related
• Research Institution: The University of Tokyo
• Principal Investigator: Yoshimi Kazuto 東京大学, 医科学研究所, 講師 (50709813)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: CRISPR-Cas3 / ゲノム編集 / 受精卵 / マウス

Outline of Final Research Achievements

We have previously reported that the Type I-E CRISPR-Cas3 system can induce a large deletion in human genome. In this research project, we examined the knockout efficiency in fertilized eggs of mice and rats as well as human cells and also investigated several modification of this system to improve its usefulness as an in vivo genome editing tool.
We succeeded in the production of Cas3 and Cascade protein complex and found that it showed higher genome editing efficiency than plasmids. We also succeeded in generating knockout mice using CRISPR-Cas3 and demonstrated its usefulness as a genome editing tool in vivo.

Free Research Field

実験動物学

Academic Significance and Societal Importance of the Research Achievements

In vivoにおける自由自在なゲノム改変技術は、ライフサイエンス研究に欠かすことのできない技術になりつつある一方、オフターゲットの影響、モザイク個体、配列特異性といった技術的問題に加えて、アメリカ主導で開発されたことによる知的財産の問題点が挙げられる。本研究で日本発ゲノム編集ツールのin vivoにおける有用性を示すことができ、新規疾患モデル動物の開発のみならず、ヒト疾患への応用に向けたゲノム編集治療戦略に大きく貢献できることが期待される。