A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-α from <i>Raphanus sativus</i> L. Plants
Human interferon (IFN) is a type of cytokine that regulates the immune system’s response to viral and bacterial infections. Recombinant IFN-α has been approved for use in the treatment of a variety of viral infections as well as an anticancer medication for various forms of leukemia. The objective o...
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MDPI AG
2022-08-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/9/8/381 |
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| author | Rashad Kebeish Emad Hamdy Omar Al-Zoubi Talaat Habeeb Raha Osailan Yassin El-Ayouty |
| author_facet | Rashad Kebeish Emad Hamdy Omar Al-Zoubi Talaat Habeeb Raha Osailan Yassin El-Ayouty |
| author_sort | Rashad Kebeish |
| collection | DOAJ |
| description | Human interferon (IFN) is a type of cytokine that regulates the immune system’s response to viral and bacterial infections. Recombinant IFN-α has been approved for use in the treatment of a variety of viral infections as well as an anticancer medication for various forms of leukemia. The objective of the current study is to produce a functionally active recombinant human IFN-α2a from transgenic <i>Raphanus sativus</i> L. plants. Therefore, a binary plant expression construct containing the IFN-α2a gene coding sequence, under the regulation of the cauliflower mosaic virus 35SS promoter, was established. <i>Agrobacterium</i>-mediated floral dip transformation was used to introduce the IFN-α2a expression cassette into the nuclear genome of red and white rooted <i>Raphanus sativus</i> L. plants. From each genotype, three independent transgenic lines were established. The anticancer and antiviral activities of the partially purified recombinant IFN-α2a proteins were examined. The isolated IFN-α2a has been demonstrated to inhibit the spread of the Vesicular Stomatitis Virus (VSV). In addition, cytotoxicity and cell apoptosis assays against Hep-G2 cells (Human Hepatocellular Carcinoma) show the efficacy of the generated IFN-α2a as an anticancer agent. In comparison to bacterial, yeast, and animal cell culture systems, the overall observed results demonstrated the efficacy of using <i>Raphanus sativus</i> L. plants as a safe, cost-effective, and easy-to-use expression system for generating active human IFN-α2a. |
| first_indexed | 2024-03-09T10:01:49Z |
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| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-09T10:01:49Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Bioengineering |
| spelling | doaj.art-ae7b03eb00334c1191558d7bb6060d242023-12-01T23:25:14ZengMDPI AGBioengineering2306-53542022-08-019838110.3390/bioengineering9080381A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-α from <i>Raphanus sativus</i> L. PlantsRashad Kebeish0Emad Hamdy1Omar Al-Zoubi2Talaat Habeeb3Raha Osailan4Yassin El-Ayouty5Botany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig 44519, EgyptBotany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig 44519, EgyptBiology Department, Faculty of Science Yanbu, Taibah University, Yanbu El-Bahr 46423, Saudi ArabiaBiology Department, Faculty of Science Yanbu, Taibah University, Yanbu El-Bahr 46423, Saudi ArabiaBiology Department, Faculty of Science Yanbu, Taibah University, Yanbu El-Bahr 46423, Saudi ArabiaBotany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig 44519, EgyptHuman interferon (IFN) is a type of cytokine that regulates the immune system’s response to viral and bacterial infections. Recombinant IFN-α has been approved for use in the treatment of a variety of viral infections as well as an anticancer medication for various forms of leukemia. The objective of the current study is to produce a functionally active recombinant human IFN-α2a from transgenic <i>Raphanus sativus</i> L. plants. Therefore, a binary plant expression construct containing the IFN-α2a gene coding sequence, under the regulation of the cauliflower mosaic virus 35SS promoter, was established. <i>Agrobacterium</i>-mediated floral dip transformation was used to introduce the IFN-α2a expression cassette into the nuclear genome of red and white rooted <i>Raphanus sativus</i> L. plants. From each genotype, three independent transgenic lines were established. The anticancer and antiviral activities of the partially purified recombinant IFN-α2a proteins were examined. The isolated IFN-α2a has been demonstrated to inhibit the spread of the Vesicular Stomatitis Virus (VSV). In addition, cytotoxicity and cell apoptosis assays against Hep-G2 cells (Human Hepatocellular Carcinoma) show the efficacy of the generated IFN-α2a as an anticancer agent. In comparison to bacterial, yeast, and animal cell culture systems, the overall observed results demonstrated the efficacy of using <i>Raphanus sativus</i> L. plants as a safe, cost-effective, and easy-to-use expression system for generating active human IFN-α2a.https://www.mdpi.com/2306-5354/9/8/381<i>Raphanus sativus</i> L.recombinant IFN-α2aapoptosisantiviral and antitumor activity |
| spellingShingle | Rashad Kebeish Emad Hamdy Omar Al-Zoubi Talaat Habeeb Raha Osailan Yassin El-Ayouty A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-α from <i>Raphanus sativus</i> L. Plants Bioengineering <i>Raphanus sativus</i> L. recombinant IFN-α2a apoptosis antiviral and antitumor activity |
| title | A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-α from <i>Raphanus sativus</i> L. Plants |
| title_full | A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-α from <i>Raphanus sativus</i> L. Plants |
| title_fullStr | A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-α from <i>Raphanus sativus</i> L. Plants |
| title_full_unstemmed | A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-α from <i>Raphanus sativus</i> L. Plants |
| title_short | A Biotechnological Approach for the Production of Pharmaceutically Active Human Interferon-α from <i>Raphanus sativus</i> L. Plants |
| title_sort | biotechnological approach for the production of pharmaceutically active human interferon α from i raphanus sativus i l plants |
| topic | <i>Raphanus sativus</i> L. recombinant IFN-α2a apoptosis antiviral and antitumor activity |
| url | https://www.mdpi.com/2306-5354/9/8/381 |
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