ORIGINAL RESEARCH article
Front. Synth. Biol.
Sec. Ecosystems and Biodiversity Sustainability
Volume 3 - 2025 | doi: 10.3389/fsybi.2025.1619871
This article is part of the Research TopicSystems Biology and Synthetic Approaches in Achieving Sustainable ActionView all articles
pTARGEX Vectors: A Versatile Toolbox for Plant-Based Protein Expression
Provisionally accepted
Allyson Marie MacLean*
Camille Chartrand-Pleau
Emily Gallipeau
Ananya Jain
Victor BoddyPatrick FakhouriSarah RehmaniTeagan ThomasMichelle Drapeau
Jordan T VanderBurgtArnaud Boudigou
Adam M Damry- University of Ottawa, Ottawa, Canada
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The ability to express heterologous proteins in specific plant tissues and subcellular compartments is critical for advancing plant-based biotechnologies and biopharming research. Here, we present the 'pTARGEX' series, a versatile toolkit designed to enable targeted protein expression within the leaves and roots of plants. These vectors incorporate subcellular targeting sequences to direct proteins of interest to the apoplast (pTARGEX-APO), cytoplasm (pTARGEX-CYT), chloroplast (pTARGEX-CHL), endoplasmic reticulum (pTARGEX-ER), or vacuole (pTARGEX-VAC). Streamlining workflow, the pTARGEX vectors encode an mCherry dropout cassette with a SapI restriction enzyme site to enable red/white screening in Escherichia coli, and an eGFP reporter gene that is constitutively expressed in planta to serve as an inbuilt marker for monitoring of successful transformation. This innovative vector series offers users an accessible, versatile platform for plant-based heterologous protein production, facilitating research and applications in systems biology, synthetic biology, and plant biotechnology.
Keywords: biopharming, Gibson assembly, GoldenGate, transgenic plants, MolecularFarming, transient expression, N. benthamiana
Received: 28 Apr 2025; Accepted: 15 Aug 2025.
Copyright: © 2025 MacLean, Chartrand-Pleau, Gallipeau, Jain, Boddy, Fakhouri, Rehmani, Thomas, Drapeau, VanderBurgt, Boudigou and Damry. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Allyson Marie MacLean, University of Ottawa, Ottawa, Canada
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