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2021 Fiscal Year Final Research Report

Improvement of genome-editing technologies by engineering of CRISPR-Cas9

Research Project

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Project/Area Number 18K19284
Research Category

Grant-in-Aid for Challenging Research (Exploratory)

Allocation TypeMulti-year Fund
Review Section Medium-sized Section 43:Biology at molecular to cellular levels, and related fields
Research InstitutionThe University of Tokyo

Principal Investigator

Nishimasu Hiroshi  東京大学, 先端科学技術研究センター, 教授 (00467044)

Project Period (FY) 2018-06-29 – 2022-03-31
KeywordsCRISPR
Outline of Final Research Achievements

Cas9 is an RNA-guided DNA nuclease derived from prokaryotic CRISPR-Cas adaptive immune systems, and associates with a guide RNA to cleave double-stranded DNA targets. Therefore, Cas9 is widely used as a powerful genome-editing tool. In this study, we determined the 3D structures of Cas9 and Cas12f, which provide mechanistic insights into their RNA-guide DNA cleavage. Based on the structural information, we further engineered Cas9 variants with improved functionality, thereby extending the utility of CRISPR-based genome engineering technologies.

Free Research Field

構造生命科学

Academic Significance and Societal Importance of the Research Achievements

本研究では、小型Cas9の結晶構造および小型Cas12fのクライオ電子顕微鏡構造を決定した。これらの立体構造から、CRISPR-Cas酵素の多様なRNA認識機構、DNA認識機構、DNA切断機構が原子レベルで明らかになった。さらに、得られた構造情報を利用し、標的範囲の拡張したCas9改変体の開発にも成功した。これらの成果は、ゲノム編集技術の高度化に貢献するものである。

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Published: 2023-01-30  

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