Plant Molecular Biology

, Volume 97, Issue 1–2, pp 103–112 | Cite as

Expression of VGRNb-PE immunotoxin in transplastomic lettuce (Lactuca sativa L.)

  • Malihe Mirzaee
  • Mokhtar Jalali-Javaran
  • Ahmad Moieni
  • Sirous Zeinali
  • Mahdi Behdani


Key message

This research has shown, for the first time, that plant chloroplasts are a suitable compartment for synthesizing recombinant immunotoxins and the transgenic immunotoxin efficiently causes the inhibition of VEGFR2 overexpression, cell growth and proliferation.


Angiogenesis refers to the formation of new blood vessels, which resulted in the growth, invasion and metastasis of cancer. The vascular endothelial growth factor receptor 2 (VEGFR2) plays a major role in angiogenesis and blocking of its signaling inhibits neovascularization and tumor metastasis. Immunotoxins are promising therapeutics for targeted cancer therapy. They consist of an antibody linked to a protein toxin and are designed to specifically kill the tumor cells. In our previous study, VGRNb-PE immunotoxin protein containing anti-VEGFR2 nanobody fused to the truncated form of Pseudomonas exotoxin A has been established. Here, we expressed this immunotoxin in lettuce chloroplasts. Chloroplast genetic engineering offers several advantages, including high levels of transgene expression, multigene engineering in a single transformation event and maternal inheritance of the transgenes. Site specific integration of transgene into chloroplast genomes, and homoplasmy were confirmed. Immunotoxin levels reached up to 1.1% of total soluble protein or 33.7 µg per 100 mg of leaf tissue (fresh weight). We demonstrated that transgenic immunotoxin efficiently causes the inhibition of VEGFR2 overexpression, cell growth and proliferation. These results indicate that plant chloroplasts are a suitable compartment for synthesizing recombinant immunotoxins.


Angiogenesis Immunotoxin Chloroplast transformation Lettuce 



The authors gratefully thank Dr. Babak Latif for creation of pBL102 vector, Dr. Mostafa Modarresi for assistance in creating pBL VGRNb-PE, Reza Moazzami for assistance with protein purification, and Ehsan Ali-Rahimi for help with the cell culture experiments.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Malihe Mirzaee
    • 1
  • Mokhtar Jalali-Javaran
    • 1
  • Ahmad Moieni
    • 1
  • Sirous Zeinali
    • 2
  • Mahdi Behdani
    • 3
  1. 1.Department of Plant Breeding & Biotechnology, Faculty of AgricultureTarbiat Modares UniversityTehranIran
  2. 2.Department of Molecular MedicinePasteur Institute of IranTehranIran
  3. 3.Biotechnology Research Center, Biotechnology Department, Venom & Biotherapeutics Molecules Lab.Pasteur Institute of IranTehranIran

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