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Poster

Session: Session 2 Presenting Authors at Posters (Odds)

P-273 - Development of a tightly regulated copper-inducible transient gene expression system in Nicotiana benthamiana incorporating a suicide exon and Cre recombinase

Wednesday, July 16, 2025
13:30 - 15:15  
Location: Confex Hall 3
Other

    Presenting Author(s)

  • BC

    Bing-Jen Chiang

    Academia Sinica
    Taipei, Taipei, Taiwan (Republic of China)

    • Author(s)

  • KL

    Kuan-Yu Lin

    Institute of Plant and Microbial Biology, Academia Sinica
    Taipei, Taipei, Taiwan (Republic of China)

  • YC

    Yi-Feng Chen

    Academia Sinica
    Taipei, Nankang, TAIWAN

  • CH

    Ching-Yi Huang

    Max Planck Institute for Biology
    Reutlingen, Baden-Wurttemberg, Germany

  • FG

    Foong-Jing Goh

    Academia Sinica
    Taipei City, Taipei, Taiwan (Republic of China)

  • LH

    Lo-Ting Huang

    Department of Plant Pathology, National Chung Hsing University
    Taichung, Taichung, Taiwan (Republic of China)

  • LC

    Li-Hung Chen

    Assistant professor
    National Chung Hsing University (NCHU), Department of Plant Pathology
    Taichung, Changhua, Taiwan (Republic of China)

  • CW

    Chih-Hang Wu

    Associate Research Fellow
    Institute of Plant and Microbial Biology, Academia Sinica
    Taipei City, Taipei, Taiwan (Republic of China)

Chemical-inducible gene expression systems are commonly used to regulate gene expression for functional genomics in various plant species. However, a convenient system that can tightly regulate transgene expression in Nicotiana benthamiana is still lacking. In this study, we developed a tightly regulated copper-inducible system that can control transgene expression and conduct cell death assays in N. benthamiana. We tested several chemical-inducible systems using Agrobacterium-mediated transient expression and found that the copper-inducible system exhibited the least concerns regarding leakiness in N. benthamiana. Although the copper-inducible system can control the expression of some tested reporters, it is not sufficiently tight to regulate certain tested hypersensitive cell death responses. Using the MoClo-based synthetic biology approach, we incorporated the suicide exon HyP5SM/OsL5 and Cre/LoxP as additional regulatory elements to enhance the tightness of the regulation. This new design allowed us to tightly control the hypersensitive cell death induced by several tested leucine-rich repeat-containing proteins and their matching avirulence factors, and it can be easily applied to regulate the expression of other transgenes in transient expression assays. Our findings offer new approaches for both fundamental and translational studies in plant functional genomics.