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2022 Fiscal Year Final Research Report

Development of an artificial light-responsive DNA repair enzyme toward deep light-dependent gene therapy

Research Project

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Project/Area Number 21K19223
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 InstitutionOsaka University

Principal Investigator

Yamamoto Junpei  大阪大学, 大学院基礎工学研究科, 准教授 (90571084)

Project Period (FY) 2021-07-09 – 2023-03-31
KeywordsDNA修復 / DNA損傷 / 人工酵素 / 機能改変
Outline of Final Research Achievements

In this study, I aimed to develop an artificial DNA repair enzyme bearing blue fluorescent protein (BFP) as a partial structure within the whole architecture toward the functional expansion of a DNA repairing enzyme photolyase, responsible for restoration of intact bipyrimidine nucleobases from ultraviolet-induced DNA damage using blue light.
After the screening of the functional enzyme using bacteria followed by the in-vitro evaluations, I finally obtained an artificial BFP-fused photolyase with 2.5-fold higher activity than the wild-type enzyme. Contrary to my initial assumption, however, the enhancement of the activity has been achieved not via the energy transfer from the fluorophore moiety in BFP. Further investigations will be required to address the molecular mechanism of the functional enhancement.

Free Research Field

光生物

Academic Significance and Societal Importance of the Research Achievements

DNA修復に関与するタンパク質群の遺伝的な欠失は様々な遺伝子疾患の原因となる。したがって、DNA修復活性を向上させた人工酵素は、これら遺伝子疾患の遺伝子治療法となる可能性を秘めているため、基礎科学の発展のみならず医学や工学への応用研究の礎となる。
今回、当初目的としていた機構に従う人工酵素は得られなかった一方で、酵素の部分構造を改変することで酵素機能が向上することを見出している。今後その構造を明らかにすることで酵素構造と活性の相関を取ることができ、酵素活性を自在にデザインする第一歩となることが期待される。

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Published: 2024-01-30  

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