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Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors

Plant-based transient expression systems have recognized potential for use as rapid and cost-effective alternatives to expression systems based on bacteria, yeast, insect, or mammalian cells. The free-floating aquatic plants of the Lemnaceae family (duckweed) have compact architecture and can be veg...

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Autores principales: Peterson, Anton, Kishchenko, Olena, Zhou, Yuzhen, Vasylenko, Maksym, Giritch, Anatoli, Sun, Jian, Borisjuk, Nikolai, Kuchuk, Mykola
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8600122/
https://www.ncbi.nlm.nih.gov/pubmed/34805101
http://dx.doi.org/10.3389/fbioe.2021.761073
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author Peterson, Anton
Kishchenko, Olena
Zhou, Yuzhen
Vasylenko, Maksym
Giritch, Anatoli
Sun, Jian
Borisjuk, Nikolai
Kuchuk, Mykola
author_facet Peterson, Anton
Kishchenko, Olena
Zhou, Yuzhen
Vasylenko, Maksym
Giritch, Anatoli
Sun, Jian
Borisjuk, Nikolai
Kuchuk, Mykola
author_sort Peterson, Anton
collection PubMed
description Plant-based transient expression systems have recognized potential for use as rapid and cost-effective alternatives to expression systems based on bacteria, yeast, insect, or mammalian cells. The free-floating aquatic plants of the Lemnaceae family (duckweed) have compact architecture and can be vegetatively propagated on low-cost nutrient solutions in aseptic conditions. These features provide an economically feasible opportunity for duckweed-based production of high-value products via transient expression of recombinant products in fully contained, controlled, aseptic and bio-safe conditions in accordance with the requirements for pharmaceutical manufacturing and environmental biosafety. Here, we demonstrated Agrobacterium-mediated high-yield transient expression of a reporter green fluorescent protein using deconstructed vectors based on potato virus X and sweet potato leaf curl virus, as well as conventional binary vectors, in two representatives of the Lemnaceae (Spirodela polyrhiza and Landoltia punctata). Aseptically cultivated duckweed populations yielded reporter protein accumulation of >1 mg/g fresh biomass, when the protein was expressed from a deconstructed potato virus X-based vector, which is capable of replication and cell-to-cell movement of the replicons in duckweed. The expression efficiency demonstrated here places duckweed among the most efficient host organisms for plant-based transient expression systems, with the additional benefits of easy scale-up and full containment.
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spelling pubmed-86001222021-11-19 Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors Peterson, Anton Kishchenko, Olena Zhou, Yuzhen Vasylenko, Maksym Giritch, Anatoli Sun, Jian Borisjuk, Nikolai Kuchuk, Mykola Front Bioeng Biotechnol Bioengineering and Biotechnology Plant-based transient expression systems have recognized potential for use as rapid and cost-effective alternatives to expression systems based on bacteria, yeast, insect, or mammalian cells. The free-floating aquatic plants of the Lemnaceae family (duckweed) have compact architecture and can be vegetatively propagated on low-cost nutrient solutions in aseptic conditions. These features provide an economically feasible opportunity for duckweed-based production of high-value products via transient expression of recombinant products in fully contained, controlled, aseptic and bio-safe conditions in accordance with the requirements for pharmaceutical manufacturing and environmental biosafety. Here, we demonstrated Agrobacterium-mediated high-yield transient expression of a reporter green fluorescent protein using deconstructed vectors based on potato virus X and sweet potato leaf curl virus, as well as conventional binary vectors, in two representatives of the Lemnaceae (Spirodela polyrhiza and Landoltia punctata). Aseptically cultivated duckweed populations yielded reporter protein accumulation of >1 mg/g fresh biomass, when the protein was expressed from a deconstructed potato virus X-based vector, which is capable of replication and cell-to-cell movement of the replicons in duckweed. The expression efficiency demonstrated here places duckweed among the most efficient host organisms for plant-based transient expression systems, with the additional benefits of easy scale-up and full containment. Frontiers Media S.A. 2021-11-04 /pmc/articles/PMC8600122/ /pubmed/34805101 http://dx.doi.org/10.3389/fbioe.2021.761073 Text en Copyright © 2021 Peterson, Kishchenko, Zhou, Vasylenko, Giritch, Sun, Borisjuk and Kuchuk. https://creativecommons.org/licenses/by/4.0/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) and the copyright owner(s) 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.
spellingShingle Bioengineering and Biotechnology
Peterson, Anton
Kishchenko, Olena
Zhou, Yuzhen
Vasylenko, Maksym
Giritch, Anatoli
Sun, Jian
Borisjuk, Nikolai
Kuchuk, Mykola
Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_full Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_fullStr Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_full_unstemmed Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_short Robust Agrobacterium-Mediated Transient Expression in Two Duckweed Species (Lemnaceae) Directed by Non-replicating, Replicating, and Cell-to-Cell Spreading Vectors
title_sort robust agrobacterium-mediated transient expression in two duckweed species (lemnaceae) directed by non-replicating, replicating, and cell-to-cell spreading vectors
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8600122/
https://www.ncbi.nlm.nih.gov/pubmed/34805101
http://dx.doi.org/10.3389/fbioe.2021.761073
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