Growing Uses of 2A in Plant Biotechnology

Luke, Garry A.; Roulston, Claire; Tilsner, Jens; Ryan, Martin

doi:10.5772/59878

Cited by 13 publications

(13 citation statements)

References 150 publications

(194 reference statements)

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“…Additionally, multimeric protein subunits or different independent proteins can be expressed and assembled in planta using a single ORF (sORF) through linkage to the 2A peptide from foot-and-mouth virus (Luke et al 2015) or inteins (interspacing polypeptide blocks of functional protein parts) (Evans et al 2005). The 2A strategy is largely used for producing important proteins, such as a multi-epitope and low-cost candidate vaccine against cysticercosis using the so-called Helios2A polyprotein system .…”

Section: Expression Cassette Collectionmentioning

confidence: 99%

Optimization of inside and outside factors to improve recombinant protein yield in plant

Habibi

Prado

Pelegrini³

et al. 2017

Plant Cell Tiss Organ Cult

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an improvement of critical features, including transcription, translation and post-translational modifications, in plant cells could make plant systems a safe and economical alternative for biopharmaceutical production. Hence, in this review, the most recent advances influencing the upstream and downstream processes involved in recombinant protein accumulation in plant cells are described. We also discuss how plant systems are becoming the benchmark for the production of several biopharmaceuticals.Keywords Heterologous expression · Host systems · Post-translational modifications · Plant-based pharmaceuticals · Recombinant protein identification and characterization of a promising regulatory pathway for the large-scale production of biopharmaceuticals could strongly contribute to the benefits of this system (Fischer et al. 2012). Molecular pharming technology presents several advantages, including (a) the production of low-cost biomass; (b) end-products lacking human toxicity; (c) the accumulation of complex proteins with correct and proper folding; and (d) straightforward methods for protein purification (Moustafa et al. 2015). Furthermore, molecular pharming offers a flexible, scalable and diverse alternative method for producing new, Abstract The use of plant systems as factories for recombinant protein production became a prominent alternative for pharmaceutical industries due to their high potential for protein accumulation. In the last decades, the application of plants for protein production has gained more attention, as plants represent an economic strategy that leads to high levels of purified and active proteins for the pharmaceutical sector. Currently, FDA approval of the first generation of recombinant proteins produced in carrot cells, taliglucerase alfa, demonstrated that plant cells have a significant capacity to express complex proteins for therapeutic use. Although plant systems still have technical and economic barriers that require improvements in future years, the optimization of upstream and downstream components affecting protein accumulation is considered a key feature in the development of new pharmaceutical proteins. Therefore,

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Section: Expression Cassette Collectionmentioning

confidence: 99%

Optimization of inside and outside factors to improve recombinant protein yield in plant

Habibi

Prado

Pelegrini³

et al. 2017

Plant Cell Tiss Organ Cult

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“…Besides the above mentioned linkers, currently, the most popular type of linker is the self-cleaving 2A peptide. For example, a sequence coding for a foot-and-mouth disease virus (FMDV) self-cleaving 2A (F2A) peptide has been used to link two genes to generate a synthetic polyprotein in plants, which could produce individual functional proteins by 2A peptide undergoing self-cleaving mechanism ( Halpin et al, 1999 ; Ralley et al, 2004 ; Luke et al, 2015 ). The F2A peptide has 19 amino acids while some other 2A peptides have 18–22 amino acids.…”

Section: Introductionmentioning

confidence: 99%

An Effective Way of Producing Fully Assembled Antibody in Transgenic Tobacco Plants by Linking Heavy and Light Chains via a Self-Cleaving 2A Peptide

et al. 2018

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Therapeutic monoclonal antibodies (mAbs) have evolved into an important class of effective medicine in treatment of various diseases. Since the antibody molecule consists of two identical heavy chains (HC) and two light chains (LC), each chain encoded by two different genes, their expressions at similar levels are critical for efficient assembly of functional recombinant mAbs. Although the plant-based expression system has been tested to produce fully assembled recombinant mAbs, coordinately expressing HC and LC at similar levels in a transgenic plant remains a challenge. A sequence coding for a foot-and-mouth disease virus (FMDV) 2A peptide has been successfully used to link two or more genes, which enable the translated polyprotein to be “self-cleaved” into individual protein in various genetically modified organisms. In the present study, we exploited the usage of F2A in Ebola virus monoclonal antibody (EBOV mAb) production. We found that transgenic tobacco plants carrying a transcription unit containing HC and LC linked by 2A not only produced similar levels of HC and LC but also rendered a higher yield of fully assembled EBOV mAb compared to those expressing HC and LC in two independent transcription units. Purified EBOV mAb bound to an Ebola epitope peptide with apparent Kd-values of 90.13–149.2 nM, indicating its proper assembly and high affinity binding to Ebola epitope peptide. To our knowledge, this is the first report showing mAb production by overexpressing a single transcription unit consisting of HC, LC and 2A in stable transformed tobacco plants.

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“…F2A is the most widely used 2A sequence in plant biotechnology and has been used to target multiple proteins to various subcellular compartments [13,34,45,46], to improve disease resistance [47,48], drought-resistance [49] and nutritional value through metabolome engineering [10,50]. Vitamin A deficiency (VAD) is a major global health issue which affects hundreds of millions of people.…”

Section: Biotechnological and Biomedical Applicationsmentioning

confidence: 99%

“…The Foot-and-mouth disease virus (FMDV) 2A sequence (hereafter "F2A") mediates "self-processing" by a novel translational effect variously referred to as 'ribosome skipping' , 'stop-go' and 'stop-carry on' translation [10][11][12]. 2A peptide cleavage has been studied in various cell types using various recombinant polyproteins and artificial reporter polyprotein systems comprising chloramphenicol acetyltransferase (CAT), β-glucuronidase (GUS), and fluorescent proteins (FPs e.g.…”

mentioning

confidence: 99%

“…Importantly, the co-translational nature of this cleavage means that by including various signal sequences within 2A polyproteins, either up-or downstream of 2A, proteins can be either targeted co-translationally to the exocytic pathway or post-translationally to different cellular compartments [34][35][36]. Of the many 2A peptides identified to date [10,27,37], four have been widely used in biotechnology and biomedicine: F2A, equine rhinitis A virus ("E2A"), porcine teschovirus-1 ("P2A") and Thosea asigna virus ("T2A") ( Table 1) [2][3][4]36,[38][39][40][41][42][43].…”

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confidence: 99%

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2A - The Ã¢ÂÂgo-toÃ¢ÂÂ Technology for Transgene Co-expression

Minskaia¹,

Luke²

2015

Single Cell Biol

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Growing Uses of 2A in Plant Biotechnology

Cited by 13 publications

References 150 publications

Optimization of inside and outside factors to improve recombinant protein yield in plant

Optimization of inside and outside factors to improve recombinant protein yield in plant

An Effective Way of Producing Fully Assembled Antibody in Transgenic Tobacco Plants by Linking Heavy and Light Chains via a Self-Cleaving 2A Peptide

2A - The Ã¢ÂÂgo-toÃ¢ÂÂ Technology for Transgene Co-expression

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Growing Uses of 2A in Plant Biotechnology

Cited by 13 publications

References 150 publications

Optimization of inside and outside factors to improve recombinant protein yield in plant

Optimization of inside and outside factors to improve recombinant protein yield in plant

An Effective Way of Producing Fully Assembled Antibody in Transgenic Tobacco Plants by Linking Heavy and Light Chains via a Self-Cleaving 2A Peptide

2A - The Ã¢ÂÂgo-toÃ¢ÂÂ Technology for Transgene Co-expression

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2A - The Ã¢ÂÂgo-toÃ¢ÂÂ Technology for Transgene Co-expression