Plant Molecular Farming
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ZMapp in an HIV context

It was truly a pleasure to run into Kevin Whaley of Mapp BioPharmaceutical today, here at the HIVR4P inernational conferrence in Cape Town - so I made him come and have coffee with me and Anna-Lise...
Ed Rybicki's insight:

I will also review a couple of molecular farming posters, so stay tuned.

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Plant Molecular Farming
A news site for the activities of members of the International Society for Plant Molecular Farming, as well as for papers of interest in the field
Curated by Ed Rybicki
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Your current "Scooper" is...Jennifer Stander

Your current "Scooper" is...Jennifer Stander | Plant Molecular Farming | Scoop.it
Hi to all the curious minds that peruse the internet for interesting things to read and enlighten your mind. I will be highjacking this page for a bit and hope to keep you all entertained! 
I am Dr Jennifer Stander, a postdoctoral research fellow in Prof Ed Rybicki's research group the Biopharming Research Unit at the University of Cape Town, South Africa. My research is focused on the development of diagnostic reagents and vaccines in plants as an alternative to mammalian and insect cells, in the hope of making these products affordable for low-to-middle-income countries where the need for these are high. 
Besides my love for science, I am also a wannabe foodie, an average baker and sewist. I always enjoy a good glass (or two!) of wine and being South African - nothing beats a proper braai.
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Glyco-Engineering Plants to Produce Helminth Glycoproteins as Prospective Biopharmaceuticals: Recent Advances, Challenges and Future Prospects

Glyco-Engineering Plants to Produce Helminth Glycoproteins as Prospective Biopharmaceuticals: Recent Advances, Challenges and Future Prospects | Plant Molecular Farming | Scoop.it

Glycoproteins are the dominant category among approved biopharmaceuticals, indicating their importance as therapeutic proteins. Glycoproteins are decorated with carbohydrate structures (or glycans) in a process called glycosylation. Glycosylation is a post-translational modification that is present in all kingdoms of life, albeit with differences in core modifications, terminal glycan structures, and incorporation of different sugar residues. Glycans play pivotal roles in many biological processes and can impact the efficacy of therapeutic glycoproteins. The majority of biopharmaceuticals are based on human glycoproteins, but non-human glycoproteins, originating from for instance parasitic worms (helminths), form an untapped pool of potential therapeutics for immune-related diseases and vaccine candidates. The production of sufficient quantities of correctly glycosylated putative therapeutic helminth proteins is often challenging and requires extensive engineering of the glycosylation pathway. Therefore, a flexible glycoprotein production system is required that allows straightforward introduction of heterologous glycosylation machinery composed of glycosyltransferases and glycosidases to obtain desired glycan structures. The glycome of plants creates an ideal starting point for N- and O-glyco-engineering of helminth glycans. Plants are also tolerant toward the introduction of heterologous glycosylation enzymes as well as the obtained glycans. Thus, a potent production platform emerges that enables the production of recombinant helminth proteins with unusual glycans. In this review, we discuss recent advances in plant glyco-engineering of potentially therapeutic helminth glycoproteins, challenges and their future prospects."

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Modified Gene Editing Systems: Diverse Bioengineering Tools and Crop Improvement

Modified Gene Editing Systems: Diverse Bioengineering Tools and Crop Improvement | Plant Molecular Farming | Scoop.it
Gene-editing systems have emerged as bioengineering tools in recent years. Classical gene-editing systems include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR) with CRISPR-associated protein 9 (Cas9) (CRISPR/Cas9), and these tools allow specific sequences to be targeted and edited. Various modified gene-editing systems have been established based on classical gene-editing systems. Base editors (BEs) can accurately carry out base substitution on target sequences, while prime editors (PEs) can replace or insert sequences. CRISPR systems targeting mitochondrial genomes and RNA have also been explored and established. Multiple gene-editing techniques based on CRISPR/Cas9 have been established and applied to genome engineering. Modified gene-editing systems also make transgene-free plants more readily available. In this review, we discuss the modifications made to gene-editing systems in recent years and summarize the capabilities, deficiencies, and applications of these modified gene-editing systems. Finally, we discuss the future developmental direction and challenges of modified gene-editing systems.
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Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants

Production of Potyvirus-Derived Nanoparticles Decorated with a Nanobody in Biofactory Plants | Plant Molecular Farming | Scoop.it
Viral nanoparticles (VNPs) have recently attracted attention for their use as building blocks for novel materials to support a range of functions of potential interest in nanotechnology and medicine. Viral capsids are ideal for presenting small epitopes by inserting them at an appropriate site on the selected coat protein (CP). VNPs presenting antibodies on their surfaces are considered highly promising tools for therapeutic and diagnostic purposes. Due to their size, nanobodies are an interesting alternative to classic antibodies for surface presentation. Nanobodies are the variable domains of heavy-chain (VHH) antibodies from animals belonging to the family Camelidae, which have several properties that make them attractive therapeutic molecules, such as their small size, simple structure, and high affinity and specificity. In this work, we have produced genetically encoded VNPs derived from two different potyviruses—the largest group of RNA viruses that infect plants—decorated with nanobodies. We have created a VNP derived from zucchini yellow mosaic virus (ZYMV) decorated with a nanobody against the green fluorescent protein (GFP) in zucchini (Cucurbita pepo) plants. As reported for other viruses, the expression of ZYMV-derived VNPs decorated with this nanobody was only made possible by including a picornavirus 2A splicing peptide between the fused proteins, which resulted in a mixed population of unmodified and decorated CPs. We have also produced tobacco etch viru
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Transient recombinant expression of highly immunogenic CagA, VacA and NapA in Nicotiana benthamiana

"Interest in the plant-based transient production of recombinant immunogenic antigens has tremendously progressed because plants are cost-effective, easily selectable, free of mammalian contamination, and support complex post-translational modifications. Nicotiana benthamiana is a convenient system for transient expression of recombinant antigens. The present study documented a platform for rapid production of Helicobacter pylori CagA, VacA and NapA antigens three days (first harvest, FH) and six days (second harvest, SH) after agro-infiltration using a syringe. In this study, CagA, VacA and NapA antigen genes from Helicobacter pylori were cloned into the binary vector pBI121 and transformed into Nicotiana benthamiana by the Agrobacterium-mediated process. Leaves of four to five weeks old Nicotiana benthamiana plants were agroinfiltrated with EHA105 subtype of Agrobacterium tumefaciens strain containing cloned CagA (pBI121-CagA), VacA (pBI121-VacA) and NapA (pBI121-NapA) constructs. The transient expression and accumulation of the recombinant genes containing CagA, VacA and NapA expression cassettes were confirmed using qRT-PCR by comparing the relative expression at FH and SH post-infiltration with the non-infiltrated (control) samples and using ELISA at 1/5 and 1/10 dilution ratios. The qRT-PCR findings showed that Agrobacterium-mediated syringe infiltration of leaves of four to five weeks old Nicotiana benthamiana plants produced significantly higher transcript levels of CagA (about 8-fold and 7-fold), VacA (38-fold and 24-fold) and NapA (7-fold and 5-fold) genes at FH and SH compared to the control sample. Besides, the maximum amount of CagA, VacA and NapA antigens were detected at the FH stage compared to the SH stage, when the antibody concentrations of the agro-infiltrated leaf extracts containing these recombinant antigens were diluted in a 1/5 ratio. This study has developed evidence to support that recombinant CagA, VacA and NapA can be transiently produced in Nicotiana benthamiana plants."

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CRISPR/Cas9-mediated knockout of the DCL2 and DCL4 genes in Nicotiana benthamiana and its productivity of recombinant proteins

Key message DCL2 and DCL4 genes in Nicotiana benthamiana plants were successfully edited using the CRISPR/Cas9 system. Abstract Recently, plants have been utilized for recombinant protein production similar to other expression systems, i.e., bacteria, yeast, insect, and mammal cells. However, insufficient amounts of recombinant proteins are often produced in plant cells. The repression of RNA silencing within plant cells could improve production levels of recombinant protein because RNA silencing frequently decomposes mRNAs from transgenes. In this study, the genes dicer-like protein 2 and 4 (NbDCL2 and NbDCL4) were successfully edited to produce double-knockout transgenic Nicotiana benthamiana plants (dcl2dcl4 plants) using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology. A transient green fluorescent protein (GFP) gene expression assay revealed that the dcl2dcl4 plants accumulated higher amounts of GFP and GFP mRNA than wild type (WT) and RNA-dependent RNA polymerase 6-knockout N. benthamiana plants (ΔRDR6 plants). Small RNA sequencing also showed that dcl2dcl4 plants accumulated lower amounts of small interfering RNAs (siRNAs) against the GFP gene than WT plants. The dcl2dcl4 plants might also produce higher amounts of human fibroblast growth factor 1 (FGF1) than WT and ΔRDR6 plants. These observations appear to reflect differences between DCLs and RDR6 in plant cell biological mechanisms. These results reveal that dcl2dcl4 plants would be suitable as platform plants for recombinant protein production.
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Frontiers | Editorial: Plant-Production Platforms for Veterinary Biopharmaceuticals | Plant Science

Public health initiatives such as 'One Health' demonstrate the growing awareness that animal and human health are strongly interconnected with each other and with the environment. Veterinary diseases are not only responsible for animal suffering and major economic losses to the livestock industry as well as a threat for endangered species but, when caused by zoonotic pathogens, may also represent a direct danger to human health as demonstrated by the recent COVID-19 pandemic (Fisher and Murray, 2021). Biopharmaceuticals such as vaccines as well as efficient diagnostic assays are the most suitable interventions to prevent and monitor the dissemination of pathogens in both domestic and wild animal populations. However, innovation is needed in this field to increase the efficacy/efficiency of these tools, simplifying delivery and use and reducing manufacturing costs. A wider utilization of novel veterinary vaccines amongst livestock/poultry/fish farmers and producers would have a major impact not only in disease prevention but also in the reduction of the use of chemicals/antibiotics for treatment of many diseases. The number of approved veterinary biotech biopharmaceuticals, mainly vaccines, is still modest when compared to that of analogous products for human use. An important issue hampering the transfer of innovative "medicines" to the veterinary field mainly relates to the generally high manufacturing costs of cutting-edge products that impact on the selling pric
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An Efficient Hairy Root System for Validation of Plant Transformation Vector and CRISPR/Cas Construct Activities in Cucumber (Cucumis sativus L.) 

An Efficient Hairy Root System for Validation of Plant Transformation Vector and CRISPR/Cas Construct Activities in Cucumber (Cucumis sativus L.)  | Plant Molecular Farming | Scoop.it

Hairy root induction system has been applied in various plant species as an effective method to study gene expression and function due to its fast-growing and high genetic stability. Recently, these systems have shown to be an effective tool to evaluate activities of CRISPR/Cas9 systems for genome editing. In this study, Rhizobium rhizogenes mediated hairy root induction was optimized to provide an effective tool for validation of plant transformation vector, CRISPR/Cas9 construct activities as well as selection of targeted gRNAs for gene editing in cucumber (Cucumis sativus L.). Under the optimized conditions including OD650 at 0.4 for infection and 5 days of co-cultivation, the highest hairy root induction frequency reached 100% for the cucumber variety Choka F1. This procedure was successfully utilized to overexpress a reporter gene (gus) and induce mutations in two Lotus japonicus ROOTHAIRLESS1 homolog genes CsbHLH66 and CsbHLH82 using CRISPR/Cas9 system. For induced mutation, about 78% of transgenic hairy roots exhibited mutant phenotypes including sparse root hair and root hair-less. The targeted mutations were obtained in individual CsbHLH66, CsbHLH82, or both CsbHLH66 and CsbHLH82 genes by heteroduplex analysis and sequencing. The hairy root transformation system established in this study is sufficient and potential for further research in genome editing of cucumber as well as other cucumis plants.

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Medicago's homegrown, plant-based COVID-19 vaccine approved by Health Canada | CBC News

Medicago's homegrown, plant-based COVID-19 vaccine approved by Health Canada | CBC News | Plant Molecular Farming | Scoop.it
Medicago's plant-based COVID-19 vaccine is now approved by Health Canada, which will soon give Canadians the option of getting a homegrown shot against SARS-CoV-2.
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Researchers awarded slice of £10million Government funds for vaccines development

Researchers awarded slice of £10million Government funds for vaccines development | Plant Molecular Farming | Scoop.it

A collaborative research team has been awarded a share of UK Government funding for research into vaccines to combat potential epidemics in developing countries. The award of nearly £500,000 to the group led by John Innes Centre virologist Professor George Lomonossoff will fund an innovative project that aims to tackle the problem of how RNA-based vaccines can be delivered to developing countries where refrigeration to low temperatures is difficult or impossible. The project includes fellow Norwich Research Park company Leaf Expression Systems and the University of Leeds and is one of 22 research projects funded by the Government’s UK Vaccine Network (UKVN) and administered by Innovate UK. The research projects will help tackle diseases which primarily affect lower income countries, including. viruses such as Ebola, Lassa Fever and Zika. Professor Lomonossoff said: “We are delighted that our project has been awarded this timely and much needed funding. The current Covid-19 pandemic has highlighted the need to protect populations from emerging disease threats. One method of achieving this is through RNA vaccines which are designed to stimulate immunity within the body.” “The major drawback of the approach, particularly in less developed regions of the world, is that RNA is inherently unstable, and vaccines based on it must be stored and distributed at low temperature. There is therefore a need to develop alternative methods for stabilising RNA molecules. Our technology provides an efficient means of storing and distributing RNA vaccines in low-and middle-income countries.” The John Innes Centre-led team will build on pioneering technology developed by Professor Lomonossoff’s group which uses Virus-Like Particles, authentic mimics of a real virus, to stimulate an immune response. These VLPs are expressed in plants to create valuable pharmaceutical products such as vaccines safely and inexpensively. The project, which begins in April 2022, will investigate how RNA molecules for vaccine use can be stabilised inside VLPs and then delivered to mammalian cells. The project aims to establish this approach as a way of addressing a range of diseases and provide a rapid response to emerging threats. In addition to known and emerging threats some of the projects announced by the UK government are also looking at ways to tackle ‘Disease X’ – a hypothetical future pathogen – to ensure the world is equipped for future epidemics or pandemics. The UKVN has funded 78 projects with over £115 million worth of UK aid funding, as part of the government’s commitment to defeat poverty, tackle instability and create prosperity in developing countries. Health and Social Care Secretary Sajid Javid said: “COVID-19 has shown us first-hand just how important it is that we work together to keep everyone across the world safe. “I am delighted that these innovative projects – tackling serious and deadly diseases – will receive the funding they need to take their research to the next stage. “Thank you to the expert scientists behind these vital projects for their efforts that will continue to save millions of lives.”

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Frontiers | Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites | Plant Science

Frontiers | Plant-Derived Cell-Free Biofactories for the Production of Secondary Metabolites | Plant Science | Plant Molecular Farming | Scoop.it
Cell-free expression systems enable the production of proteins and metabolites within a few hours or days. Removing the cellular context while maintaining the protein biosynthesis apparatus provides an open system that allows metabolic pathways to be installed and optimized by expressing different numbers and combinations of enzymes. This facilitates the synthesis of secondary metabolites that are difficult to produce in cell-based systems because they are toxic to the host cell or immediately converted into downstream products. Recently, we developed a cell-free lysate derived from tobacco BY-2 cell suspension cultures for the production of recombinant proteins. This system is remarkably productive, achieving yields of up to 3 mg/mL in a one-pot in vitro transcription–translation reaction and contains highly active energy and cofactor regeneration pathways. Here, we demonstrate for the first time that the BY-2 cell-free lysate also allows the efficient production of several classes of secondary metabolites. As case studies, we synthesized lycopene, indigoidine, betanin, and betaxanthins, which are useful in the food, cosmetic, textile, and pharmaceutical industries. Production was achieved by the co-expression of up to three metabolic enzymes. For all four products, we achieved medium to high yields. However, the yield of betanin (555 μg/mL) was outstanding, exceeding the level reported in yeast cells by a factor of more than 30. Our results show that the BY-2 cell-fre
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Engineering a Plant-Derived Astaxanthin Synthetic Pathway Into Nicotiana benthamiana 

Engineering a Plant-Derived Astaxanthin Synthetic Pathway Into Nicotiana benthamiana  | Plant Molecular Farming | Scoop.it
Carotenoids have been shown to be essential for human nutrition. Consumption of carotenoid-rich fruits and vegetables can reduce the risk of many diseases. The ketocarotenoid astaxanthin has become a commercially valuable compound due to its powerful antioxidant properties compared to other carotenoids. It is naturally produced in certain algae, bacteria, and the flowers of some species of the genus Adonis, although it is produced in such small quantities in these organisms that it is costly to extract. Chemical synthesis of this compound has also shown limited success with a high proportion of esterified forms of astaxanthin being produced, which decreases antioxidant properties by the conversion of hydroxyl groups to esters. Previously, transgenic astaxanthin-producing plants have been created using a β-carotene ketolase enzyme of either bacterial or algal origin. However, a novel astaxanthin pathway exists in the flowering plants of the genus Adonis which has not been utilized in the same manner. The pathway involves two unique enzymes, β-ring-4-dehydrogenase and 4-hydroxy-β-ring-4-dehydrogenase, which add the necessary hydroxyl and ketone groups to the rings of β-carotene. In the present study, Nicotiana benthamiana plants were transformed with chimeric constructs coding for these two enzymes. The regenerated, transgenic plants accumulate astaxanthin and their growth (height and weight) was unaffected, when compared to non-transformed N. benthamiana and to plant
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Highly Efficient Generation of Canker-Resistant Sweet Orange Enabled by an Improved CRISPR/Cas9 System 

Highly Efficient Generation of Canker-Resistant Sweet Orange Enabled by an Improved CRISPR/Cas9 System  | Plant Molecular Farming | Scoop.it
Sweet orange (Citrus sinensis) is the most economically important species for the citrus industry. However, it is susceptible to many diseases including citrus bacterial canker caused by Xanthomonas citri subsp. citri (Xcc) that triggers devastating effects on citrus production. Conventional breeding has not met the challenge to improve disease resistance of sweet orange due to the long juvenility and other limitations. CRISPR-mediated genome editing has shown promising potentials for genetic improvements of plants. Generation of biallelic/homozygous mutants remains difficult for sweet orange due to low transformation rate, existence of heterozygous alleles for target genes, and low biallelic editing efficacy using the CRISPR technology. Here, we report improvements in the CRISPR/Cas9 system for citrus gene editing. Based on the improvements we made previously [dicot codon optimized Cas9, tRNA for multiplexing, a modified sgRNA scaffold with high efficiency, citrus U6 (CsU6) to drive sgRNA expression], we further improved our CRISPR/Cas9 system by choosing superior promoters [Cestrum yellow leaf curling virus (CmYLCV) or Citrus sinensis ubiquitin (CsUbi) promoter] to drive Cas9 and optimizing culture temperature. This system was able to generate a biallelic mutation rate of up to 89% for Carrizo citrange and 79% for Hamlin sweet orange. Consequently, this system was used to generate canker-resistant Hamlin sweet orange by mutating the effector binding element (EBE) o
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 The B1 Domain of Streptococcal Protein G Serves as a Multi-Functional Tag for Recombinant Protein Production in Plants

 The B1 Domain of Streptococcal Protein G Serves as a Multi-Functional Tag for Recombinant Protein Production in Plants | Plant Molecular Farming | Scoop.it
Plants have long been considered a cost-effective platform for recombinant production. A recently recognized additional advantage includes the low risk of contamination of human pathogens, such as viruses and bacterial endotoxins. Indeed, a great advance has been made in developing plants as a “factory” to produce recombinant proteins to use for biopharmaceutical purposes. However, there is still a need to develop new tools for recombinant protein production in plants. In this study, we provide data showing that the B1 domain of Streptococcal protein G (GB1) can be a multi-functional domain of recombinant proteins in plants. N-terminal fusion of the GB1 domain increased the expression level of various target proteins ranging from 1.3- to 3.1-fold at the protein level depending on the target proteins. GB1 fusion led to the stabilization of the fusion proteins. Furthermore, the direct detection of GB1-fusion proteins by the secondary anti-IgG antibody eliminated the use of the primary antibody for western blot analysis. Based on these data, we propose that the small GB1 domain can be used as a versatile tag for recombinant protein production in plants.
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Lettuce Could Protect Astronauts’ Bones on Mars Trip

Lettuce Could Protect Astronauts’ Bones on Mars Trip | Plant Molecular Farming | Scoop.it
Astronauts might one day grow and eat genetically modified plants to ward off disease associated with long spaceflights. Researchers at the University of California, Davis, College of Engineering have developed a transgenic, or genetically modified, lettuce producing a drug to protect against bone density loss in microgravity. The work will be presented March 22 at the spring meeting of the American Chemical Society in San Diego.
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Agrobacterium-Mediated Genetic Transformation of Embryogenic Callus in a Liriodendron Hybrid (L. Chinense × L. Tulipifera)

Agrobacterium-Mediated Genetic Transformation of Embryogenic Callus in a Liriodendron Hybrid (L. Chinense × L. Tulipifera) | Plant Molecular Farming | Scoop.it
A highly efficient genetic transformation system of Liriodendron hybrid embryogenic calli through Agrobacterium-mediated genetic transformation was established and optimized. The Agrobacterium tumefaciens strain EHA105, harboring the plasmid pBI121, which contained the ß-glucuronidase (GUS) gene and neomycin phosphotransferase II (npt II) gene under the control of the CaMV35S promoter, was used for transformation. Embryogenic calli were used as the starting explant to study several factors affecting the Agrobacterium-mediated genetic transformation of the Liriodendron hybrid, including the effects of various media, selection by different Geneticin (G418) concentrations, pre-culture period, Agrobacterium optical density, infection duration, co-cultivation period, and delayed selection. Transformed embryogenic calli were obtained through selection on medium containing 90 mg L−1 G418. Plant regeneration was achieved and selected via somatic embryogenesis on medium containing 15 mg L−1 G418. The optimal conditions included a pre-culture time of 2 days, a co-culture time of 3 days, an optimal infection time of 10 min, and a delayed selection time of 7 days. These conditions, combined with an OD600 value of 0.6, remarkably enhanced the transformation rate. The results of GUS chemical tissue staining, polymerase chain reaction (PCR), and southern blot analysis demonstrated that the GUS gene was successfully expressed and integrated into the Liriodendron hybrid genome.
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Production of recombinant β-glucocerebrosidase in wild-type and glycoengineered transgenic Nicotiana benthamiana root cultures with different N-glycan profiles

Production of recombinant β-glucocerebrosidase in wild-type and glycoengineered transgenic Nicotiana benthamiana root cultures with different N-glycan profiles | Plant Molecular Farming | Scoop.it

"Gaucher disease is an inherited lysosomal storage disorder caused by an insufficiency of active β-glucocerebrosidase (GCase). Exogenous recombinant GCase via enzyme replacement therapy is considered the most practical treatment for Gaucher disease. Mannose receptors mediate the efficient uptake of exogenous GCase into macrophages. Thus, terminal mannose residues on N-glycans are essential for the delivery of exogenous GCase. In this study, recombinant GCase was produced in root cultures of wild-type (WT) and glycoengineered transgenic Nicotiana benthamiana with downregulated N-acetylglucosaminyltransferase I expression. Root cultures of WT and glycoengineered transgenic N. benthamiana plants were successfully generated by the induction of plant hormones. Recombinant GCases produced in both root cultures possessed GCase enzyme activity. Purified GCases derived from both root cultures revealed different N-glycan profiles. The WT-derived GCase possessed the predominant plant-type N-glycans, which contain plant-specific sugars-linkages, specifically β1,2-xylose and α1,3-fucose residues. Notably, the mannosidic-type N-glycans with terminal mannose residues were abundant in the purified GCase derived from glycoengineered N. benthamiana root culture. This research provides a promising plant-based system for the production of recombinant GCase with terminal mannose residues on N-glycans."

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Reproducibility and flexibility of monoclonal antibody production with Nicotiana benthamiana

Reproducibility and flexibility of monoclonal antibody production with Nicotiana benthamiana | Plant Molecular Farming | Scoop.it
The ongoing SARS-CoV-2 coronavirus pandemic of 2020–2021 underscores the need for manufacturing platforms that can rapidly produce monoclonal antibody (mAb) therapies. As reported here, a platform based on Nicotiana benthamiana produced mAb therapeutics with high batch-to-batch reproducibility and flexibility, enabling production of 19 different mAbs of sufficient purity and safety for clinical application(s). With a single manufacturing run, impurities were effectively removed for a representative mAb product (the ZMapp component c4G7). Our results show for the first time the reproducibility of the platform for production of multiple batches of clinical-grade mAb, manufactured under current Good Manufacturing Practices, from Nicotiana benthamiana. The flexibility of the system was confirmed by the results of release testing of 19 different mAbs generated with the platform. The process from plant infection to product can be completed within 10 days. Therefore, with a constant supply of plants, response to the outbreak of an infectious disease could be initiated within a matter of weeks. Thus, these data demonstrated that this platform represents a reproducible, flexible system for rapid production of mAb therapeutics to support clinical development.
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Producing Consistent, High Quality mAbs with Plants

Producing Consistent, High Quality mAbs with Plants | Plant Molecular Farming | Scoop.it
Resources Producing Consistent, High Quality mAbs with Plants iBio’s new paper, “Quality Plant-Made mAbs with FastPharming®,” illustrates FastPharming’s ability to manufacture consistent, high-quality antibodies.
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Modifying Anthocyanins Biosynthesis in Tomato Hairy Roots: A Test Bed for Plant Resistance to Ionizing Radiation and Antioxidant Properties in Space 

Modifying Anthocyanins Biosynthesis in Tomato Hairy Roots: A Test Bed for Plant Resistance to Ionizing Radiation and Antioxidant Properties in Space  | Plant Molecular Farming | Scoop.it
Gene expression manipulation of specific metabolic pathways can be used to obtain bioaccumulation of valuable molecules and desired quality traits in plants. A single-gene approach to impact different traits would be greatly desirable in agrospace applications, where several aspects of plant physiology can be affected, influencing growth. In this work, MicroTom hairy root cultures expressing a MYB-like transcription factor that regulates the biosynthesis of anthocyanins in Petunia hybrida (PhAN4), were considered as a testbed for bio-fortified tomato whole plants aimed at agrospace applications. Ectopic expression of PhAN4 promoted biosynthesis of anthocyanins, allowing to profile 5 major derivatives of delphinidin and petunidin together with pelargonidin and malvidin-based anthocyanins, unusual in tomato. Consistent with PhAN4 features, transcriptomic profiling indicated upregulation of genes correlated to anthocyanin biosynthesis. Interestingly, a transcriptome reprogramming oriented to positive regulation of cell response to biotic, abiotic, and redox stimuli was evidenced. PhAN4 hairy root cultures showed the significant capability to counteract reactive oxygen species (ROS) accumulation and protein misfolding upon high-dose gamma irradiation, which is among the most potent pro-oxidant stress that can be encountered in space. These results may have significance in the engineering of whole tomato plants that can benefit space agriculture.
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Frontiers | Metabolic Engineering of Crocin Biosynthesis in Nicotiana Species | Plant Science

Frontiers | Metabolic Engineering of Crocin Biosynthesis in Nicotiana Species | Plant Science | Plant Molecular Farming | Scoop.it
Crocins are high-value soluble pigments that are used as colorants and supplements, their presence in nature is extremely limited and, consequently, the high cost of these metabolites hinders their use by other sectors, such as the pharmaceutical and cosmetic industries. The carotenoid cleavage dioxygenase 2L (CsCCD2L) is the key enzyme in the biosynthetic pathway of crocins in Crocus sativus. In this study, CsCCD2L was introduced into Nicotiana tabacum and Nicotiana glauca for the production of crocins. In addition, a chimeric construct containing the Brevundimonas sp. β-carotene hydroxylase (BrCrtZ), the Arabidopsis thaliana ORANGE mutant gene (AtOrMut), and CsCCD2L was also introduced into N. tabacum. Quantitative and qualitative studies on carotenoids and apocarotenoids in the transgenic plants expressing CsCCD2L alone showed higher crocin level accumulation in N. glauca transgenic plants, reaching almost 400 μg/g DW in leaves, while in N. tabacum 36 μg/g DW was obtained. In contrast, N. tabacum plants coexpressing CsCCD2L, BrCrtZ, and AtOrMut accumulated, 3.5-fold compared to N. tabacum plants only expressing CsCCD2L. Crocins with three and four sugar molecules were the main molecular species in both host systems. Our results demonstrate that the production of saffron apocarotenoids is feasible in engineered Nicotiana species and establishes a basis for the development of strategies that may ultimately lead to the commercial exploitation of these valuable pigment
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Online workshop on: Plant-made pharmaceuticals for mucosal administration - Register Now, for FREE!

Online workshop on: Plant-made pharmaceuticals for mucosal administration - Register Now, for FREE! | Plant Molecular Farming | Scoop.it
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Plant-made products for the win!

Plant-made products for the win! | Plant Molecular Farming | Scoop.it
AzarGen Biotechnologies became one of only a few South African biotechs to be granted a pre-IND meeting with the US-FDA and to receive important guidance for manufacturing, pre-clinical and clinical development of our plant-made rituximab, AZ2000™, earmarked for further development to treat Rheumatoid Arthritis and eventual use in Non-Hodgkin’s Lymphoma (NHL patients). The AZ2000™ type-B meeting briefing package comprised of manufacturing, quality, in vitro, toxicology and efficacy data, were generated and compiled during the two years of international COVID-19 pandemic restrictions. This achievement was supported and enabled by AzarGen’s development partners: iBio, Inc., Envol Biomedical and CBR International, and funding from South Africa’s Industrial Development Corporation. The AzarGen management team, Mauritz Venter and Cobus Zwiegelaar consider feedback from the US-FDA as a major milestone, which serves as a stepping stone towards sharing knowledge and lessons learned with the South African biotech and drug development community.
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Benthamiana rhapsody

Benthamiana rhapsody | Plant Molecular Farming | Scoop.it
How benthi — this wild tobacco plant from the Australian outback — became a model experimental system that is accelerating research and saving lives. If you’re a natural history buff like I am, you…
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 Editorial: Plant Science's Contribution to Fighting Viral Pandemics: COVID-19 as a Case Study | Plant Science

 Editorial: Plant Science's Contribution to Fighting Viral Pandemics: COVID-19 as a Case Study | Plant Science | Plant Molecular Farming | Scoop.it
The use of plants to treat human diseases can be traced back to 6000 years ago. The enormous biochemical arsenal of plants known as the secondary (or specialized) metabolome has enabled the production of medicinal compounds to cure and/or ameliorate symptoms of many human and animal diseases, that are still prevalent today. The enormous progress experienced by plant biotechnology in the last thirty years further boosted the potential of plants to cope with diseases thanks to the advancement of molecular pharming, i.e. the use of plants to produce, by transient or stable transformation, biopharmaceuticals.Given the current pandemic caused by the SARS-CoV-2, the international scientific community has gathered together in an urgent search for solutions to the current COVID-19 outbreak. So far, the plant-derived compounds chloroquine phosphate and artemisinin, previously used against malaria, are being repurposed for the treatment of COVID-19. While Medicago is developing a candidate vaccine in plant factories, based on Virus-Like Particles (VLPs) of SARS-CoV-2, that is currently in Phase III clinical trial and it holds promises to be one of the first subunit vaccines to enter the market. Furthermore, ongoing research supports plants as bio-factories for the production of eagerly-awaited SARS-CoV-2 monoclonal antibodies for passive immunization meant to manage the disease.In this emergency scenario, our topic aims to gather all the relevant information in plant sciences t
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Frontiers | Utilizing Plant Synthetic Biology to Improve Human Health and Wellness | Plant Science

Frontiers | Utilizing Plant Synthetic Biology to Improve Human Health and Wellness | Plant Science | Plant Molecular Farming | Scoop.it
Plants offer a vast source of bioactive chemicals with the potential to improve human health through the prevention and treatment of disease. However, many potential therapeutics are produced in small amounts or in species that are difficult to cultivate. The rapidly evolving field of plant synthetic biology provides tools to capitalize on the inventive chemistry of plants by transferring metabolic pathways for therapeutics into far more tenable plants, increasing our ability to produce complex pharmaceuticals in well-studied plant systems. Plant synthetic biology also provides methods to enhance the ability to fortify crops with nutrients and nutraceuticals. In this review, we discuss (1) the potential of plant synthetic biology to improve human health by generating plants that produce pharmaceuticals, nutrients, and nutraceuticals and (2) the technological challenges hindering our ability to generate plants producing health-promoting small molecules.
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