論文

査読有り 国際誌
2020年8月20日

I-SceI Endonuclease-Mediated Plant Genome Editing by Protein Transport through a Bacterial Type III Secretion System.

Plants (Basel, Switzerland)
  • Yuki Yanagawa
  • ,
  • Kasumi Takeuchi
  • ,
  • Masaki Endo
  • ,
  • Ayako Furutani
  • ,
  • Hirokazu Ochiai
  • ,
  • Seiichi Toki
  • ,
  • Ichiro Mitsuhara

9
9
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.3390/plants9091070

Xanthomonas campestris is one of bacteria carrying a type III secretion system which transports their effector proteins into host plant cells to disturb host defense system for their infection. To establish a genome editing system without introducing any foreign gene, we attempted to introduce genome editing enzymes through the type III secretion system. In a test of protein transfer, X. campestris pv. campestris (Xcc) transported a considerable amount of a reporter protein sGFP-CyaA into tobacco plant cells under the control of the type III secretion system while maintaining cell viability. For proof of concept for genome editing, we used a reporter tobacco plant containing a luciferase (LUC) gene interrupted by a meganuclease I-SceI recognition sequence; this plant exhibits chemiluminescence of LUC only when a frameshift mutation is introduced at the I-SceI recognition site. Luciferase signal was observed in tobacco leaves infected by Xcc carrying an I-SceI gene which secretes I-SceI protein through the type III system, but not leaves infected by Xcc carrying a vector control. Genome-edited tobacco plant could be regenerated from a piece of infected leaf piece by repeated selection of LUC positive calli. Sequence analysis revealed that the regenerated tobacco plant possessed a base deletion in the I-SceI recognition sequence that activated the LUC gene, indicating genome editing by I-SceI protein transferred through the type III secretion system of Xcc.

リンク情報
DOI
https://doi.org/10.3390/plants9091070
PubMed
https://www.ncbi.nlm.nih.gov/pubmed/32825459
PubMed Central
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7570400
ID情報
  • DOI : 10.3390/plants9091070
  • PubMed ID : 32825459
  • PubMed Central 記事ID : PMC7570400

エクスポート
BibTeX RIS