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Benchmarking genetic elements for high-level protein expression using a novel Marchantia polymorpha transient

Xin Yan1, Bowen Peng1, Zhiqi Miao1

  • 1Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.

International Journal of Biological Macromolecules
|March 30, 2026
PubMed
Summary

Researchers developed a rapid Agrobacterium-mediated transient expression system for Marchantia polymorpha, enabling quick characterization of genetic elements like promoters and terminators to improve synthetic biology applications.

Keywords:
Gene expression elementsMarchantia polymorphaTransient system

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Area of Science:

  • Plant synthetic biology
  • Molecular biology
  • Genetics

Background:

  • Marchantia polymorpha is a key model for plant synthetic biology.
  • Characterizing genetic elements is crucial for predictable gene expression.
  • Limited genetic parts and reliance on stable transformation hinder Marchantia research.

Purpose of the Study:

  • To develop a rapid Agrobacterium-mediated transient expression system for Marchantia.
  • To systematically benchmark genetic elements for gene expression.
  • To expand the genetic toolkit for Marchantia and enhance protein yields.

Main Methods:

  • Developed an 8-day workflow for Agrobacterium-mediated transient expression.
  • Systematically evaluated 21 promoters, 15 terminators, and 7 signal peptides.
  • Validated transient system results with stable transformation assays.

Main Results:

  • Identified top-performing elements: P_35S-3 promoter, T_MpAct1 terminator, and SP_SdMir signal peptide.
  • P_35S-3 promoter showed a 409-fold increase in activity compared to P_35S.
  • Achieved high eGFP protein yield (319.8 μg/g fresh weight) using P_35S-3 in stable lines.

Conclusions:

  • The transient expression system is rapid and efficient for characterizing gene expression elements.
  • This system expands the genetic toolkit for Marchantia.
  • Enhanced protein expression levels are achievable in Marchantia using identified elements.