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Updated: Jun 3, 2025

Transient Gene Expression in Tobacco using Gibson Assembly and the Gene Gun
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Modeling coding sequence design for virus-based expression in tobacco.

Moritz Burghardt1, Tamir Tuller1,2,3

  • 1Department of Biomedical Engineering, The Iby and Aladar Fleischman Faculty of Engineering, Tel Aviv, Israel.

Synthetic and Systems Biotechnology
|January 13, 2025
PubMed
Summary
This summary is machine-generated.

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Optimizing recombinant protein production in tobacco plants requires careful gene design. This study reveals that codon usage, translational ramps, and amino acid composition significantly impact protein yield, offering practical guidelines for enhanced expression.

Area of Science:

  • Plant biotechnology
  • Molecular biology
  • Biochemistry

Background:

  • Transient expression in tobacco is a common method for recombinant protein production.
  • Agrobacterium-mediated delivery enables high protein yields, but optimizing heterologous gene expression remains challenging.
  • Adapting coding sequences to the host has yielded inconsistent results.

Purpose of the Study:

  • To investigate the impact of various sequence features on recombinant protein yield in tobacco.
  • To develop predictive models for optimizing gene expression in plants.
  • To provide practical guidelines for coding sequence design.

Main Methods:

  • Compiled a dataset of over 200 published expression studies, including coding sequences and protein yields.
Keywords:
ICONcodon usageheterologous gene expressionplantssynthetic biologytransient expressionviral based vectors

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  • Evaluated established gene expression models on a subset of the data.
  • Analyzed sequence features such as codon usage, tRNA adaptation, translational ramps, and amino acid composition.

    • Main Results:

    • Tobacco codon usage alone showed moderate predictive power for protein yield.
    • Codon usage of viruses and agrobacteria proved useful for predicting heterologous expression.
    • Key predictive features include tRNA dynamics, translational ramps, and amino acid composition.
    • A comprehensive model achieved a 0.57 correlation with protein yield.

    Conclusions:

    • Protein yield in tobacco is influenced by complex sequence features beyond simple codon adaptation.
    • Incorporating features like translational ramps and host-specific codon usage can improve predictive models.
    • This research offers practical insights for designing synthetic gene constructs for efficient plant-based protein production.