한국환경농학회지 (Korean Journal of Environmental Agriculture)

  • 제40권3호
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  • Pages.179-185
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  • 2021
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  • 1225-3537(pISSN)
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  • 2233-4173(eISSN)
한국환경농학회 (The Korean Society of Environmental Agriculture)
권호 표지
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후숙 조절 유전자 Pectate lyase와 Phytoene Synthase 편집용 CRISPR-Cas9 sgRNA의 유전자 편집 효율 측정

Evaluation of sgRNAs Targeting Pectate Lyase and Phytoene Synthase for Delaying Tomato Fruit Ripening

  • Park, Hyosun (Department of Agricultural Chemistry, Chonnam National University) ;
  • Yang, So Hee (Department of Agricultural Chemistry, Chonnam National University) ;
  • Kim, Euyeon (Department of Agricultural Chemistry, Chonnam National University) ;
  • Koo, Yeonjong (Department of Agricultural Chemistry, Chonnam National University)
  • 투고 : 2021.08.31
  • 심사 : 2021.09.18
  • 발행 : 2021.09.30
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초록

BACKGROUND: Tomato genome editing using CRISPR-Cas9 is being actively conducted in recent days, and lots of plant researches have been aiming to develop high valued crops by editing target genes without inserting foreign genes. Many researchers have been involved in the manipulation of the crop ripening process because fruit ripening is an important fruit phenotype for increasing fruit shelf life, taste, and texture of crops. This paper intends to evaluate target sgRNA to edit the two ripening-related genes encoding pectate lyase (PL) and phytoene synthase (Psy) with the CRISPR-Cas9 system. METHODS AND RESULTS: The CRISPR-Cas9 expression vector was cloned to target the PL (Solyc03g111690), Psy1 (Solyc03g031860), and Psy2 (Solyc02g081330) genes, which are the ripening genes of tomatoes. Tomatoes injected with Agrobacterium containing the CRISPR-Cas9 expression vector were further cultured for 5 days and used to check gene editing efficiency. As a result of the target gene sequence analysis by the next generation sequencing method, gene editing efficiency was calculated, and the efficient target location was selected for the PL and Psy genes. CONCLUSION: Therefore, this study was aimed to establish target sgRNA data that could have higher efficiency of the CRISPR-Cas9 system to obtain the delayed ripening phenotype of tomato. The developed method and sgRNA information is expected to be utilized in the development of various crops to manage its ripening processes.

키워드

과제정보

This work was supported by the Cooperative Research Program for Agriculture Science and Technology Development (Project No. PJ01477601) funded by Rural Development Administration.

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