Rational design of genome editing protein toward fast and accurate editing

2022 Fiscal Year Final Research Report

• Project/Area Number: 20K06571
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 43040:Biophysics-related
• Research Institution: Tohoku University
• Principal Investigator: Kamagata Kiyoto 東北大学, 多元物質科学研究所, 准教授 (90432492)
• Project Period (FY): 2020-04-01 – 2023-03-31
• Keywords: ゲノム編集 / タンパク質工学 / 単分子計測 / スライディング / 合理的設計

Outline of Final Research Achievements

Genome editing is a technology that freely rewrites DNA containing genetic information using a special DNA-binding protein. It is expected to be used in gene therapy in the future, but there are problems such as time-consuming editing and mistakenly editing non-target DNA sequences. In this study, we combined single-molecule measurement and protein engineering to modify the fast and accurate genome-editing protein Cas9 by promoting the sliding motion along the DNA. We identified sites that inhibit the sliding movement of Cas9 and sites that promote the sliding of other proteins, and modified Cas9. Single-molecule fluorescence measurements revealed that the modified Cas9 accelerated the sliding movement by about 8-fold.

Free Research Field

生物物理学

Academic Significance and Societal Importance of the Research Achievements

遺伝子治療でゲノム編集タンパク質の使用が期待されている。しかし、既存のゲノム編集タンパク質Cas9には、編集時間がかかる、標的ではないDNA配列を誤って編集するなどの問題がある。遺伝子治療への応用を考えたとき、DNAの誤編集は致命的です。本研究で改変したCas9は、DNA上を素早く移動できるため、編集にかかる時間が短くなり、誤編集も減ると考えられるため、遺伝子治療などのゲノム編集での使用が期待される。