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Plants as Molecular Farming Factories
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Plants as Molecular Farming Factories

A special issue of Plants (ISSN 2223-7747). This special issue belongs to the section "Plant Molecular Biology".

Deadline for manuscript submissions: closed (20 July 2023) | Viewed by 9423

Special Issue Editors

Prof. Dr. Elena Victorovna Deineko

E-Mail Website
Guest Editor
Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia
Interests: plant physiology; molecular farming; gene editing; CRISPR; genetic engineering
Prof. Dr. María Alejandra Alvarez

E-Mail Website
Guest Editor
Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Universidad Maimónides-CEBBAD. Hidalgo 775 (1113), CABA, Argentina
Interests: plant biotechnology; molecular farming; plant secondary metabolism; in vitro plant cell culture

Special Issue Information

Dear Colleagues,

Molecular Farming is the production of different molecules for the pharmaceutical and chemical industries in transgenic organisms and is achieved through genetic manipulation; however, it is mainly used to describe the production of recombinant proteins in plants. Some of the advantages of the production of recombinant proteins in plants are aspects related to biosecurity since plants do not produce toxins, as bacteria do, nor bear pathogens for humans and animals, as animal cells do. Additionally, plants are able to produce complex glycoproteins and folding in the same way that mammal cells are. Finally, the transient production and the in vitro culture platform are valuable tools that have previously demonstrated their utility to increase recombinant protein yields. Due to the COVID-19 pandemic, molecular farming has demonstrated its relevance in the field of recombinant vaccine production. The bioproduction of recombinant pharmaceuticals in plant systems is becoming a promising alternative to existing platforms that are based on mammalian or bacterial cells. The cultivation of plant cells under the controlled conditions of bioreactors ensures the production of high-quality proteins that are in accordance with GMP (Good Manufacturing Practice) standards. This Special Issue of Plants will include articles discussing the most recent novelties that are related to this promising area.

Prof. Dr. Elena Victorovna Deineko
Prof. Dr. María Alejandra Alvarez
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Plants is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • molecular farming
  • plant expression systems
  • transgenic plants
  • bioreactors
  • recombinant proteins
  • biopharmaceuticals

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Published Papers (4 papers)

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Research

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11 pages, 1570 KiB  
Article
Rapid Transient Expression of Receptor-Binding Domain of SARS-CoV-2 and the Conserved M2e Peptide of Influenza A Virus Linked to Flagellin in Nicotiana benthamiana Plants Using Self-Replicating Viral Vector
by Eugenia S. Mardanova, Roman Y. Kotlyarov and Nikolai V. Ravin
Plants 2022, 11(24), 3425; https://doi.org/10.3390/plants11243425 - 8 Dec 2022
Cited by 4 | Viewed by 1813
Abstract
The development of recombinant vaccines against SARS-CoV-2 and influenza A is an important task. The combination of the conserved influenza A antigen, the extracellular domain of the transmembrane protein M2 (M2e), and the receptor-binding domain of the SARS-CoV-2 spike glycoprotein (RBD) provides the [...] Read more.
The development of recombinant vaccines against SARS-CoV-2 and influenza A is an important task. The combination of the conserved influenza A antigen, the extracellular domain of the transmembrane protein M2 (M2e), and the receptor-binding domain of the SARS-CoV-2 spike glycoprotein (RBD) provides the opportunity to develop a bivalent vaccine against these infections. The fusion of antigens with bacterial flagellin, the ligand for Toll-like receptor 5 and potent mucosal adjuvant, may increase the immunogenicity of the candidate vaccines and enable intranasal immunization. In this study, we report the transient expression of RBD alone, RBD coupled with four copies of M2e, and fusions of RBD and RBD-4M2e with flagellin in Nicotiana benthamiana plants using the self-replicating potato virus X-based vector pEff. The yields of purified recombinant proteins per gram of fresh leaf tissue were about 20 µg for RBD, 50–60 µg for RBD-4M2e and the fusion of RBD with flagellin, and about 90 µg for RBD-4M2e fused to flagellin. Targeting to the endoplasmic reticulum enabled the production of glycosylated recombinant proteins comprising RBD. Our results show that plant-produced RBD and RBD-4M2e could be further used for the development of subunit vaccines against COVID-19 and a bivalent vaccine against COVID-19 and influenza A, while flagellin fusions could be used for the development of intranasal vaccines. Full article
(This article belongs to the Special Issue Plants as Molecular Farming Factories)
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19 pages, 2535 KiB  
Article
Modulation of the Translation Efficiency of Heterologous mRNA and Target Protein Stability in a Plant System: The Case Study of Interferon-αA
by Alexander A. Tyurin, Orkhan Mustafaev, Aleksandra V. Suhorukova, Olga S. Pavlenko, Viktoriia A. Fridman, Ilya S. Demyanchuk and Irina V. Goldenkova-Pavlova
Plants 2022, 11(19), 2450; https://doi.org/10.3390/plants11192450 - 20 Sep 2022
Cited by 1 | Viewed by 1572
Abstract
A broad and amazingly intricate network of mechanisms underlying the decoding of a plant genome into the proteome forces the researcher to design new strategies to enhance both the accumulation of recombinant proteins and their purification from plants and to improve the available [...] Read more.
A broad and amazingly intricate network of mechanisms underlying the decoding of a plant genome into the proteome forces the researcher to design new strategies to enhance both the accumulation of recombinant proteins and their purification from plants and to improve the available relevant strategies. In this paper, we propose new approaches to optimize a codon composition of target genes (case study of interferon-αA) and to search for regulatory sequences (case study of 5′UTR), and we demonstrated their effectiveness in increasing the synthesis of recombinant proteins in plant systems. In addition, we convincingly show that the approach utilizing stabilization of the protein product according to the N-end rule or a new protein-stabilizing partner (thermostable lichenase) is sufficiently effective and results in a significant increase in the protein yield manufactured in a plant system. Moreover, it is validly demonstrated that thermostable lichenase as a protein-stabilizing partner not only has no negative effect on the target protein activity (interferon-αA) integrated in its sequence, but rather enhances the accumulation of the target protein product in plant cells. In addition, the retention of lichenase enzyme activity and interferon biological activity after the incubation of plant protein lysates at 65 °C and precipitation of nontarget proteins with ethanol is applicable to a rapid and inexpensive purification of fusion proteins, thereby confirming the utility of thermostable lichenase as a protein-stabilizing partner for plant systems. Full article
(This article belongs to the Special Issue Plants as Molecular Farming Factories)
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Review

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31 pages, 1103 KiB  
Review
Three Parts of the Plant Genome: On the Way to Success in the Production of Recombinant Proteins
by Sergey M. Rozov, Alla A. Zagorskaya, Yuri M. Konstantinov and Elena V. Deineko
Plants 2023, 12(1), 38; https://doi.org/10.3390/plants12010038 - 21 Dec 2022
Cited by 3 | Viewed by 2326
Abstract
Recombinant proteins are the most important product of current industrial biotechnology. They are indispensable in medicine (for diagnostics and treatment), food and chemical industries, and research. Plant cells combine advantages of the eukaryotic protein production system with simplicity and efficacy of the bacterial [...] Read more.
Recombinant proteins are the most important product of current industrial biotechnology. They are indispensable in medicine (for diagnostics and treatment), food and chemical industries, and research. Plant cells combine advantages of the eukaryotic protein production system with simplicity and efficacy of the bacterial one. The use of plants for the production of recombinant proteins is an economically important and promising area that has emerged as an alternative to traditional approaches. This review discusses advantages of plant systems for the expression of recombinant proteins using nuclear, plastid, and mitochondrial genomes. Possibilities, problems, and prospects of modifications of the three parts of the genome in light of obtaining producer plants are examined. Examples of successful use of the nuclear expression platform for production of various biopharmaceuticals, veterinary drugs, and technologically important proteins are described, as are examples of a high yield of recombinant proteins upon modification of the chloroplast genome. Potential utility of plant mitochondria as an expression system for the production of recombinant proteins and its advantages over the nucleus and chloroplasts are substantiated. Although these opportunities have not yet been exploited, potential utility of plant mitochondria as an expression system for the production of recombinant proteins and its advantages over the nucleus and chloroplasts are substantiated. Full article
(This article belongs to the Special Issue Plants as Molecular Farming Factories)
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12 pages, 1193 KiB  
Review
Genetic Containment for Molecular Farming
by Amy L. Klocko
Plants 2022, 11(18), 2436; https://doi.org/10.3390/plants11182436 - 19 Sep 2022
Cited by 2 | Viewed by 2175
Abstract
Plant molecular farming can provide humans with a wide variety of plant-based products including vaccines, therapeutics, polymers, industrial enzymes, and more. Some of these products, such as Taxol, are produced by endogenous plant genes, while many others require addition of genes by artificial [...] Read more.
Plant molecular farming can provide humans with a wide variety of plant-based products including vaccines, therapeutics, polymers, industrial enzymes, and more. Some of these products, such as Taxol, are produced by endogenous plant genes, while many others require addition of genes by artificial gene transfer. Thus, some molecular farming plants are transgenic (or cisgenic), while others are not. Both the transgenic nature of many molecular farming plants and the fact that the products generated are of high-value and specific in purpose mean it is essential to prevent accidental cross-over of molecular farming plants and products into food or feed. Such mingling could occur either by gene flow during plant growth and harvest or by human errors in material handling. One simple approach to mitigate possible transfer would be to use only non-food non-feed species for molecular farming purposes. However, given the extent of molecular farming products in development, testing, or approval that do utilize food or feed crops, a ban on use of these species would be challenging to implement. Therefore, other approaches will need to be considered for mitigation of cross-flow between molecular farming and non-molecular-farming plants. This review summarized some of the production systems available for molecular farming purposes and options to implement or improve plant containment. Full article
(This article belongs to the Special Issue Plants as Molecular Farming Factories)
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