Engineering zucchini yellow mosaic potyvirus as a non-pathogenic vector for expression of heterologous proteins in cucurbits

Arazi, Tzahi; Slutsky, Shalom Guy; Shiboleth, Yoel Moshe; Wang, Yongzeng; Rubinstein, Menachem; Barak, Sara; Yang, Jie; Gal‐On, Amit

doi:10.1016/s0168-1656(01)00229-2

Cited by 70 publications

(56 citation statements)

References 46 publications

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“…For example, we can mass-produce therapeutic proteins in their active forms in edible plants, enabling the oral-delivery of protein drugs (Arazi et al, 2001;Ma et al, 2005). Currently, heterologous genes can be expressed in plants using either transgenic plants or plant viral vectors (Matsumura et al, 2000;Menassa et al, 2000;Gils et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

Matsuo

Hong

Tabayashi

et al. 2006

Planta

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We have developed Cucumber mosaic virus (CMV) as a plant virus vector especially for production of pharmaceutical proteins. The CMV vector is a vector modifiable for different host plants and does not require further engineering steps. CMV contains three genomic RNA molecules (RNAs 1-3) necessary for infectivity. With this system, instead of creating different vector constructs for each plant we use, we take advantage of the formation of pseudrecombinants between two CMV isolates by simply reassembling a vector construct (RNA 2 base) and an RNA molecule containing the host determinant (mostly RNA 3). In this study, the gene for acidic fibroblast growth factor (aFGF), one of the human cytokines, was cloned under the control of the subgenomic promoter for RNA 4A of the CMV-based vector, C2-H1. Infected Nicotiana benthamiana plants produced aFGF at levels up to 5-8% of the total soluble protein. The tobacco-produced aFGF was purified, and its biological activity was confirmed. Using this system, which provides a versatile and viable strategy for production of therapeutic proteins in plants, we also demonstrated a high level of aFGF in Glycine max (soybean) and Arabidopsis thaliana.

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Section: Introductionmentioning

confidence: 99%

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

Matsuo

Hong

Tabayashi

et al. 2006

Planta

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“…Potyviral-based vectors expressing full-length individual proteins have been developed previously (Dolja et al, 1992;Guo et al, 1998;Choi et al, 2000;GermanRetana et al, 2000;Masuta et al, 2000;Arazi et al, 2001;Fernández-Fernández et al, 2001;Beauchemin et al, 2005). However, the utility of most constructs designed for the overproduction of proteins of interest could be limited by their genetic instability due to RNA recombination events that rapidly eliminate foreign sequences (Dolja et al, 1993;Guo et al, 1998;Choi et al, 2000;German-Retana et al, 2000).…”

Section: Discussionmentioning

confidence: 99%

“…The NIb-CP junction has also been used as the cloning site in order to obtain stable chimeras (Fernández-Fernández et al, 2001). Some authors (Arazi et al, 2001) claim that a foreign gene inserted between the NIb and CP genes is relatively less prone to recombination events than genes inserted between the P1 and HC-Pro.…”

Section: Discussionmentioning

confidence: 99%

“…In other potyviral-based vectors it was observed that uidA suffered spontaneous deletions after prolonged propagation or subsequent plant passage (Dolja et al, 1993;Guo et al, 1998;Choi et al, 2000;German-Retana et al, 2000). Although the instability of uidA may be caused by the length of the foreign sequence, some results indicate that other factors affect insert stability (Choi et al, 2000;Arazi et al, 2001;Beauchemin et al, 2005). The use of plant virus-based vectors instead of virusfree plant transformation for the production of foreign proteins presents several advantages (Scholthof et al, 1996;Arntzen, 1997), although both systems are of great value under different circumstances.…”

Section: Discussionmentioning

confidence: 99%

“…On top of the geometrical considerations, other attractive features of potyvirus-derived vectors are the fact that at least some potyviruses are infectious as cloned cDNA, thus facilitating the inoculation process, and viral expressed proteins can reach high percentages of the final protein present in the infected cells. All these traits have prompted several research groups to derive vectors form members of the Potyviruses (Dolja et al, 1992;Guo et al, 1998;Choi et al, 2000;GermanRetana et al, 2000;Masuta et al, 2000;Arazi et al, 2001;Beauchemin et al, 2005).…”

Section: Resumen Altos Niveles De Expresión De Proteínas Foráneas a Pmentioning

confidence: 99%

See 2 more Smart Citations

High expression of foreign proteins from a biosafe viral vector derived from Turnip mosaic virus

Touriño

Sánchez

Fereres

et al. 2008

Span. j. agric. res.

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A vector derived from an original infectious clone of Turnip mosaic potyvirus (TuMV) has been developed. The vector (p35Tunos-vec01) was made through the creation of a new unique insertion site for foreign genes between the NIb and the CP genes of the viral genome. The jellyfish green fluorescent protein (GFP) and E. coli beta-glucuronidase (GUS) were expressed in Arabidopsis thaliana plants infected with chimeric vectors carrying their corresponding genes. In the case of GUS, expression levels that were 15-50 fold higher than nuclear transgenic A. thaliana lines carrying the same gene were attained. Acceptable levels of gene stability were found in the infected plants, compatible with the use of the vector for protein production in plants. A non-aphid transmissible version of the vector was also made and its lack of aphid transmission was extensively shown. The vector was not transmitted through seeds or by contact between plants.

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A potyvirus vector efficiently targets recombinant proteins to chloroplasts, mitochondria and nuclei in plant cells when expressed at the amino terminus of the polyprotein

Majer

Navarro

Daròs

2015

Biotechnology Journal

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Plant virus-based expression systems allow quick and efficient production of recombinant proteins in plant biofactories. Among them, a system derived from tobacco etch virus (TEV; genus potyvirus) permits coexpression of equimolar amounts of several recombinant proteins. This work analyzed how to target recombinant proteins to different subcellular localizations in the plant cell using this system. We constructed TEV clones in which green fluorescent protein (GFP), with a chloroplast transit peptide (cTP), a nuclear localization signal (NLS) or a mitochondrial targeting peptide (mTP) was expressed either as the most amino-terminal product or embedded in the viral polyprotein. Results showed that cTP and mTP mediated efficient translocation of GFP to the corresponding organelle only when present at the amino terminus of the viral polyprotein. In contrast, the NLS worked efficiently at both positions. Viruses expressing GFP in the amino terminus of the viral polyprotein produced milder symptoms. Untagged GFPs and cTP and NLS tagged amino-terminal GFPs accumulated to higher amounts in infected tissues. Finally, viral progeny from clones with internal GFPs maintained the extra gene better. These observations will help in the design of potyvirus-based vectors able to coexpress several proteins while targeting different subcellular localizations, as required in plant metabolic engineering.

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Engineering zucchini yellow mosaic potyvirus as a non-pathogenic vector for expression of heterologous proteins in cucurbits

Cited by 70 publications

References 46 publications

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

Development of Cucumber mosaic virus as a vector modifiable for different host species to produce therapeutic proteins

High expression of foreign proteins from a biosafe viral vector derived from Turnip mosaic virus

A potyvirus vector efficiently targets recombinant proteins to chloroplasts, mitochondria and nuclei in plant cells when expressed at the amino terminus of the polyprotein

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