Development of a novel method for insect gene editing by adult injection

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K21311
• 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: Kyoto University
• Principal Investigator: Daimon Takaaki 京都大学, 農学研究科, 教授 (70451846)
• Project Period (FY): 2020-07-30 – 2022-03-31
• Keywords: ゲノム編集 / 遺伝子編集 / 昆虫 / 形質デザイン / 昆虫機能利用

Outline of Final Research Achievements

In this project, we developed a simple and accessible method for insect gene editing, termed Direct Parental CRISPR (DIPA-CRISPR). We show that injection of Cas9 ribonucleoproteins (RNPs) into the haemocoel of adult females efficiently introduces heritable mutations in developing oocytes. Importantly, commercially available standard Cas9 protein can be directly used for DIPA-CRISPR, which makes this approach highly practical and feasible. DIPA-CRISPR enables highly efficient gene editing in the cockroaches, on which conventional approaches cannot be applied, and in the model beetle Tribolium castaneum. Due to its simplicity and accessibility, DIPA-CRISPR will greatly extend the application of gene editing technology to a wide variety of insects.

Free Research Field

昆虫科学

Academic Significance and Societal Importance of the Research Achievements

昆虫科学は昆虫の特異な形態・生理・生態を理解することで将来にわたる人類の生存を保証しようとする学問分野である。本課題では、革新的な昆虫ゲノム編集法を開発に取り組み、成虫に注射することで高効率にゲノム編集を行うことができる方法（DIPA-CRISPR）を確立した。本法の開発によって、ゲノム編集ツールCas9 RNP（市販品でよい）を成虫に注射するだけ、という簡便な方法で昆虫のゲノム編集を行うことができるようになった。