Overexpression of human erythropoietin (EPO) affects plant morphologies: retarded vegetative growth in tobacco and male sterility in tobacco and Arabidopsis

Cheon, Ban Yoon; Kim, Hae Jin; Oh, Ki-Won; Bahn, Sung Chul; Ahn, Ji Hoon; Choi, Jang Won; Ok, Sung Han; Bae, Jung Myung; Shin, Jeong Sheop

doi:10.1007/s11248-004-2737-3

Cited by 59 publications

(59 citation statements)

References 36 publications

(29 reference statements)

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“…Among other tested approaches, the codon optimization provided no benefit, EPO native signal peptide was processed more efficiently compared with tobacco signal peptide, and glycosylation was very important for EPO stability in plant tissue. Contrary to our findings with LicKM fusion, others reported that fusion to the elastin-like polypeptide (ELP) carrier protein had a negligible effect on EPO accumulation, which the authors explain by a potential toxicity of the EPO-ELP fusion to the plant [76,79]. This finding is also in contradiction with the potent enhancing effect of fusion to ELP on the expression of human IL-10 and murine IL-4 [80], spider silk protein [80,81], full-size anti-HIV-1 antibody 2F5 [82] in tobacco leaves, and single-chain variable The average of four determinations is shown.…”

Section: Fig 11contrasting

confidence: 99%

“…When expressed by transgenic Physcomitrella patens lines, EPO was secreted through cell wall into culture medium and over 6 days, accumulated approximately 250 mg/g dry weight of moss [75]. In transgenic tobacco plants, high-level expression of non-fused human EPO containing its native signal peptide has been shown to affect plant morphology, causing retarded vegetative growth and male sterility [76]. However, by modifying the tobacco transformation system (eg, reducing cytokinin 6-BA concentration and changing a strain of Agrobacterium) and the expression cassette (flanking the EPO gene with the His tag and KDEL sequences), Musa et al [77] have developed transgenic tobacco overexpressing EPO with no plant health abnormality.…”

Section: Fig 11mentioning

confidence: 99%

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Plant-Produced Human Recombinant Erythropoietic Growth Factors Support Erythroid Differentiation In Vitro

Musiychuk

Sivalenka²,

Jaje

et al. 2013

Stem Cells and Development

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Clinically available red blood cells (RBCs) for transfusions are at high demand, but in vitro generation of RBCs from hematopoietic stem cells requires significant quantities of growth factors. Here, we describe the production of four human growth factors: erythropoietin (EPO), stem cell factor (SCF), interleukin 3 (IL-3), and insulin-like growth factor-1 (IGF-1), either as non-fused proteins or as fusions with a carrier molecule (lichenase), in plants, using a Tobacco mosaic virus vector-based transient expression system. All growth factors were purified and their identity was confirmed by western blotting and peptide mapping. The potency of these plant-produced cytokines was assessed using TF1 cell (responsive to EPO, IL-3 and SCF) or MCF-7 cell (responsive to IGF-1) proliferation assays. The biological activity estimated here for the cytokines produced in plants was slightly lower or within the range cited in commercial sources and published literature. By comparing EC 50 values of plant-produced cytokines with standards, we have demonstrated that all four plant-produced growth factors stimulated the expansion of umbilical cord blood-derived CD34 + cells and their differentiation toward erythropoietic precursors with the same potency as commercially available growth factors. To the best of our knowledge, this is the first report on the generation of all key bioactive cytokines required for the erythroid development in a cost-effective manner using a plant-based expression system.

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Section: Fig 11contrasting

confidence: 99%

Section: Fig 11mentioning

confidence: 99%

Plant-Produced Human Recombinant Erythropoietic Growth Factors Support Erythroid Differentiation In Vitro

Musiychuk

Sivalenka²,

Jaje

et al. 2013

Stem Cells and Development

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“…For example, constitutive expression of the human epo gene affects vegelarive growth and male sterility of transgenic tobacco (Cheon et al, 2004). In our study, few AP-transformed plants bore seeds, suggesting that high constitutive hGM-CSF expression during early seed development may have harmed fertility.…”

Section: Discuss|onmentioning

confidence: 62%

Aptamers improve the expression of a human granulocyte-macrophage colony stimulating factor in transgenicArabidopsis thaliana seeds

Wang

2007

J. Plant Biol.

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Domain IV nucleotides of human 7SL RNA comprise an approximately 50-nucleotide region with a highly conserved secondary structure consisting of two internal loops. We inserted these into a modified 3'-untranslated region of the human granulocyte-macrophage colony stimulating factor (hGM-CSF) gene. The expression vector, under the control of a tissue-specific promoter derived from the soybean ~' subunit of ~3-conglycinin, was introduced into Arabidopsis thaliana by Agrobacteriurn.

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“…Recently developed plant biotechnology offers additional advantages in production scale, economy, product safety, and ways of delivery (1)(2)(3)(4)(5)(6). At present, the main goal is to increase the overall expression yield of functional plant-derived proteins, especially those of viral origin, which sometimes seriously impair transgenic plant growth and development (42)(43)(44).…”

Section: Discussionmentioning

confidence: 99%

Smallpox subunit vaccine produced in planta confers protection in mice

Golovkin

Spitsin

Andrianov

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

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We report here the in planta production of the recombinant vaccinia virus B5 antigenic domain (pB5), an attractive component of a subunit vaccine against smallpox. The antigenic domain was expressed by using efficient transient and constitutive plant expression systems and tested by various immunization routes in two animal models. Whereas oral administration in mice or the minipig with collard-derived insoluble pB5 did not generate an anti-B5 immune response, intranasal administration of soluble pB5 led to a rise of B5-specific immunoglobulins, and parenteral immunization led to a strong anti-B5 immune response in both mice and the minipig. Mice immunized i.m. with pB5 generated an antibody response that reduced virus spread in vitro and conferred protection from challenge with a lethal dose of vaccinia virus. These results indicate the feasibility of producing safe and inexpensive subunit vaccines by using plant production systems.plant biotechnology ͉ transgenic plants ͉ B5 glycoprotein ͉ recombinant antigen P lants have emerged as an excellent alternative to other expression systems for the production of complex pharmaceutical proteins, including recombinant subunit vaccines (1-6). It was shown that some plant-derived antigens can induce systemic and mucosal immune responses and, in some cases, confer protection against challenge (1-3). Plants provide the additional advantage of direct delivery through oral or other mucosal routes (1, 6). Despite some difficulties with the expression of certain recombinant proteins, especially those of viral origin, plant biotechnology holds the promise of producing medicinal proteins to be used in vaccine formulations.Interest in a safe smallpox vaccine has been reawakened by the threat of bioterrorism (7, 8) and continuous outbreaks of orthopoxvirus diseases (9, 10). A live vaccinia virus (VV)-based vaccine has been used to eventually eradicate smallpox disease (11, 12), but does display side effects (13). Although one approach for developing a safer vaccine is to use a highly attenuated live virus, recombinant protein-based vaccines are likely to be safer. For orthopoxviruses, there are several candidate antigens that can protect mice and nonhuman primates from lethal challenge (14-20). The extracellular virus (EV)-specific membrane glycoprotein encoded by the B5R gene (21-24) is the main target of EV-neutralizing antibodies present in human-derived vaccinia gamma globulin used to treat complications arising from smallpox vaccination (25).In this study, we demonstrate that the VV B5 protein can be produced in two plant expression systems. The use of the magnifection transient expression system (26-28) enabled rapid high-yield production of soluble B5 as well as selection of optimal subcellular targeting signals to use in stable plant transformation. Preparations of purified plant-derived B5 antigen (pB5) induced a strong immune response when administered parenterally and intranasally, and mice vaccinated i.m. with pB5 were protected from VV challenge. ResultsExpression Ca...

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Overexpression of human erythropoietin (EPO) affects plant morphologies: retarded vegetative growth in tobacco and male sterility in tobacco and Arabidopsis

Cited by 59 publications

References 36 publications

Plant-Produced Human Recombinant Erythropoietic Growth Factors Support Erythroid Differentiation In Vitro

Plant-Produced Human Recombinant Erythropoietic Growth Factors Support Erythroid Differentiation In Vitro

Aptamers improve the expression of a human granulocyte-macrophage colony stimulating factor in transgenicArabidopsis thaliana seeds

Smallpox subunit vaccine produced in planta confers protection in mice

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