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Development of DNA-free genome editing technology for producing a new breed of citrus

Research Project

Project/Area Number 19K21150
Project/Area Number (Other) 18H06010 (2018)
Research Category

Grant-in-Aid for Research Activity Start-up

Allocation TypeMulti-year Fund (2019)
Single-year Grants (2018)
Review Section 0602:Agricultural and environmental biology and related fields
Research InstitutionEhime University

Principal Investigator

Kaya Hidetaka  愛媛大学, 農学研究科, 准教授 (80398825)

Project Period (FY) 2018-08-24 – 2020-03-31
Project Status Completed (Fiscal Year 2019)
Budget Amount *help
¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2018: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Keywordsゲノム編集 / カンキツ / カラタチ / DNAフリー / Cas9 / 柑橘 / CRISPR/Cas9
Outline of Research at the Start

愛媛県の最重要作物である柑橘類において,DNA を用いないゲノム編集(DNA フリー・ゲノム編集)技術の確立を目指す.この目的のために,パーティクルボンバードメント法により,ゲノム編集酵素複合体をカンキツ細胞に直接導入する.この方法により作出したゲノム編集カンキツは,遺伝子組み換え生物の規制対象外となると考えられる.よって,この技術は,カンキツの革新的な育種技術となると期待される.

Outline of Final Research Achievements

We have been researching for the purpose of developing a technology base for developing new citrus cultivars using DNA-free genome editing technology. First, we confirmed the sequence of the PDS gene of Poncirus trifoliata as the target gene, prepared sgRNA, and confirmed its function in vitro. Next, we examined the conditions for applying the particle bombardment method to the callus of P. trifoliata. After confirming the transduction efficiency using the GUS gene, RNP (Ribonucleoprotein) was injected and CAPS analysis was performed. However, so far no mutations due to genome editing have been confirmed. At present, we are also studying particles used in particle bombardment.

Academic Significance and Societal Importance of the Research Achievements

DNAフリー・ゲノム編集技術を柑橘類において確立することで,遺伝子組換え体の規制の対象外となるゲノム編集柑橘品種を作出できる.愛媛大学では,愛媛県オリジナル柑橘品種のゲノム解析が進められており,各品種における果実特性に関するゲノム情報が明らかになりつつある.DNAフリー・ゲノム編集技術を柑橘を適用することで,ゲノム情報と果実特性の関連性を検証し,圃場での特性評価実験も可能になる.本研究により柑橘類の機能ゲノム研究が進むことで,新品種や機能性柑橘の開発につながると期待される.

Report

(3 results)
  • 2019 Annual Research Report   Final Research Report ( PDF )
  • 2018 Annual Research Report

URL: 

Published: 2018-08-27   Modified: 2024-03-26  

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