Special Issue "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: 31 December 2022 | Viewed by 344

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 2200 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

Published Papers

This special issue is now open for submission, see below for planned papers.

Planned Papers

The below list represents only planned manuscripts. Some of these manuscripts have not been received by the Editorial Office yet. Papers submitted to MDPI journals are subject to peer-review.

Article

Optimization of recombinant human interferon gamma (dIFN) production in the knock-in monoclonal cell lines Arabidopsis thaliana

Tatyana V. Marenkova, Elena A. Uvarova and Elena V. Deineko

Abstract: The production of recombinant human interferon gamma in monoclonal A.thaliana cell lines with the human dIFN gene, delivered to the region of the histone H3.3 gene by the CRISPR/Cas9 method, was more than 2% of the TSP (Permyakova et al, 2021). The results of the analysis of the assessment of the recombinant target protein with changes in the composition of nutrient media and under stressful effects on cell cultures are presented. Determining the increase in biomass and the amount of dIFN protein during cultivation on different nutrient media will allow selecting the medium with the highest yield of the target protein. A further increase in the growth characteristics of cell cultures will be achieved by adding minerals, amino acids, and growth regulators to the culture medium. The effect of short-term osmotic stress upon the addition of mannitol on the production of the target protein is also considered.

 

Review

Three parts of the plant genome - on the way to success in the production of recombinant proteins

 Yurii V.Sidorchuk, Alla A.Zagorskaya, Yurii M.Konstantinov and Elena V.Deineko

Abstract: Recombinant proteins are the most important product of the modern biotechnological industry. They are indispensable in medicine for diagnostics and therapy, in the food, chemical industry and in scientific research. Plant cells combine the advantages of the eukaryotic protein production system, the simplicity and profitability of the bacterial one. The use of plants for the production of recombinant proteins is an economically significant and promising area that is developing as an alternative to traditional ones. This review discusses the advantages of plant systems for the expression of recombinant proteins using nuclear, plastid, and mitochondrial genomes. The possibilities, problems and prospects of modification of the three parts of the genome in the light of obtaining producing plants are discussed. Examples of the successful use of the nuclear expression platform for the production of various biopharmaceuticals, veterinary drugs, and technologically important proteins, as well as examples demonstrating a high yield of recombinant protein upon modification of the chloroplast genome, are presented. The prospects of using plant mitochondria as an expression system for the production of recombinant proteins, its advantages over the nucleus and chloroplasts are substantiated. Despite the fact that these possibilities have not yet been realized, examples of progress towards the delivery of exogenous DNA into the mitochondrial genome of plants are given.

 

Review

Production of recombinant proteins in plant cell culture: the role of secretory signal peptides

Elena A. Uvarova and Elena V. Deineko

Abstract: Plants and plant cells are currently attracting close attention of researchers as promising expression systems for the production of commercially important recombinant proteins. The production of recombinant proteins largely depends on the correct localization of the target protein in the plant cell. This purpose is served by special signals fused with the target protein. Such signals allow proteins to be sorted along the plant cell secretion pathways and protein to accumulate in the target compartment. This review will present an analysis of such signals and describe their molecular mechanisms. Using the experimental data obtained by researchers over the past few years, the effectiveness of using such signals for the production of recombinant proteins in plant expression systems will be demonstrated. A brief overview of bioinformatics tools for searching and analyzing suitable signal sequences will be given.

 

Article

Title: Strategies for Efficient Expression of a Heterologous Gene in a Plant System: Increasing the Translational Activity of the mRNA and the Stability of the Target Protein by the Interferon αA as the Example
Authors: Alexander A. Tyurin , Orkhan Mustafaev, Aleksandra V. Suhorukova, Viktoriia A. Fridman, and Irina V. Goldenkova‑Pavlova

Abstract: One of the main problems of the effective use of plants as producers of pharmaceutical proteins is to ensure the effectiveness of the main biological processes for the implementation of genetic information. Realization of genetic information is a multistage process, including transcription, processing of pre-mRNA, translation, and ensuring the stability of the target protein. This study: (i) evaluated the contribution of genetic determinants that are important for the efficient translation of mRNA of heterologous genes, such as 5'-untranslated regions (5'-UTR), codon usage of target genes, using new theoretical approaches; (ii) ensuring the stability of the target protein by changing the codon for the destabilizing a.o. in the second position on the codon corresponding to the stabilizing amino acid, and tested thermostable lichenase as a new protein stabilizing partner for plant systems. The obtained experimental data confirmed the advantage of using the studied regulatory codes and a protein-stabilizing partner to significantly increase the yield of the target protein in plant systems, which, depending on the scale of synthesis, can provide a significant advantage in terms of resource costs.
Highlights: Modulation of Translation Efficiency of the Heterologous mRNA and Target Protein Stability in a Plant System by the Example of Interferon αA

 

Article

Title: Expression of early secretory antigenic target 6 (ESAT-6) from Mycobacterium tuberculosis in cucumber and its immunogenicity in mice
Authors: Andreas G. Lössl; Mohammad Tahir Waheed; Kiran Saba; Muhammad Sameeullah; Johanna Gottschamel; Fatima Ijaz; Muhammad Suleman Malik; Sabeen Malik; Ekrem Gurel

Abstract: Tuberculosis is one of the deadliest infectious diseases, especially in the developing and underdeveloped countries. There is dire need of vaccine that should be affordable and provide immunity to adults from Mycobacterium strains, unlike currently used BCG vaccine, which is administered and effective in children. Plants have emerged as an advantageous and affordable platform for many pharmaceuticals and vaccines. In the present work, 6 kDa early secretory antigenic target (ESAT-6) from M. tuberculosis (MTB) was expressed in Cucumis sativus, an edible plant, through Agrobacterium-mediated transformation. Transformation and presence of transgene in transgenic plants was confirmed by polymerase chain reaction (PCR) using ESAT-6 specific primers. qRT-PCR data showed the integration of one copy of ESAT-6 gene in transgenic plants. The expression of ESAT-6 antigen was evaluated by immunoblot analysis, which was estimated to accumulate up to 1% of total soluble protein (TSP). Protein expression was further confirmed by ELISA. The recombinant ESAT-6 protein was purified from transgenic plants and detected using coomassie staining and Western blotting. ESAT-6 protein induced significant humoral immune response in mice immunized both orally and subcutaneously. In conclusion, the present study can serve to establish a platform for edible vaccine production against tuberculosis.

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