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Engineering Chlamydomonas reinhardtii for Expression of Functionally Active Human Interferon-α

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Abstract

Human interferon (IFN) are secreted cytokines that play a major regulatory role in response to various infections. Commercially, IFN-α has been approved to treat many chronic viral diseases as well as a variety of cancers and different types of leukemia. In this study, a binary vector containing human IFN-α2a gene under the regulation of the cauliflower mosaic virus 35S promoter was constructed. IFN-œ2a expression cassette was transferred to Chlamydomonas reinhardtii cells via Agrobacterium-mediated transformation method. Three independent transgenic C. reinhartii lines were generated and reported to produce a biologically active IFN-œ2a. The expressed IFN-œ2a was partially purified and tested for their antitumor and antiviral properties. Cytotoxicity and cell apoptosis assays involving the usage of the recombinant C. reinhardtii IFN-œ2a (Cr. IFN-œ2a) against the growth of Hep-G2 cells (human hepatocellular carcinoma), EAC-induced tumors (Ehrlich Ascites Carcinoma) in mice prove the functionality of the produced IFN-œ2a as an anticancer drug. Moreover, Cr.IFN-œ2a is shown to have significant inhibitory effects on the propagation of the vesicular stomatitis virus (VSV). The overall observed results support the application of C. reinhardtii expression system as a cost effective, eco-friendly, safe, and easy to employ compared to plant, bacterial and animal cell culture systems.

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Acknowledgements

This work was in part under the supervision of the Applied Scientific Research Center, Herbal and Medicinal plants research group, Taibah University. We are thankful to Dr. Ali Fahmy Head of R&D Sector at VACSERA, Egypt for his valuable help with the antitumor and antiviral assays.

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Authors and Affiliations

  1. Botany and Microbiology Department, Faculty of Science, Zagazig University, El-Gamaa Street 1, Zagazig, Sharkia, 44519, Egypt

    Yassin El-Ayouty

  2. Botany Department, Faculty of Science, Suez Canal Univeristy, Ismailia, Egypt

    Islam El-Manawy & Emad Hamdy

  3. Clinical Pathology Department, Kaser El-Aini Medical School - Cairo University, Cairo, Egypt

    Sherif Nasih

  4. Biology Department, Faculty of Science Yanbu, Taibah University, King Khalid Rd, Al amoedi, Yanbu El-Bahr, 46423, Saudi Arabia

    Rashad Kebeish

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  1. Yassin El-Ayouty
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  2. Islam El-Manawy
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  3. Sherif Nasih
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  4. Emad Hamdy
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  5. Rashad Kebeish
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Correspondence to Rashad Kebeish.

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El-Ayouty, Y., El-Manawy, I., Nasih, S. et al. Engineering Chlamydomonas reinhardtii for Expression of Functionally Active Human Interferon-α. Mol Biotechnol 61, 134–144 (2019). https://doi.org/10.1007/s12033-018-0143-y

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