Development of technology to increase protein expression by modifying 5'-untranslated region by genome editing

2022 Fiscal Year Final Research Report

• Project/Area Number: 19K22299
• Research Category: Grant-in-Aid for Challenging Research (Exploratory)
• Allocation Type: Multi-year Fund
• Review Section: Medium-sized Section 39:Agricultural and environmental biology and related fields
• Research Institution: Hokkaido University
• Principal Investigator: Onouchi Hitoshi 北海道大学, 農学研究院, 教授 (50322839)
• Project Period (FY): 2019-06-28 – 2023-03-31
• Keywords: ゲノム編集 / 5'非翻訳訳領域

Outline of Final Research Achievements

The aim of this project was to develop a method to modulate the expression level of a specific protein by using genome editing without introducing foreign DNA into the genome. To achieve this, we took the approach of cutting the 5'-leader region of the target gene by the CRISPR/Cas9 system and introducing random mutations via non-homologous end joining (NHEJ) into the 5'-leader region. Our results demonstrated that the protein expression level can be altered to varying degrees by NHEJ-mediated random mutations in the 5'-leader region.

Free Research Field

分子生物学

Academic Significance and Societal Importance of the Research Achievements

遺伝子組換え作物は有用な形質を持っていても、消費者から敬遠される傾向がある。本研究で開発した方法では、外来導入DNAを残さずに特定のタンパク質の発現量の調節が可能で、多くのタンパク質の発現調節に応用できると考えられる。そのため、遺伝子組換えに該当しない範囲のゲノム編集を用いて様々なタンパク質の発現量の改変による品種改良が可能となり、作物育種への大きな波及効果が期待できる。