Kebeish Rashad, Hamdy Emad, Al-Zoubi Omar, Habeeb Talaat, Osailan Raha, El-Ayouty Yassin
Botany and Microbiology Department, Faculty of Science, Zagazig University, Zagazig 44519, Egypt.
Biology Department, Faculty of Science Yanbu, Taibah University, Yanbu El-Bahr 46423, Saudi Arabia.
Bioengineering (Basel). 2022 Aug 10;9(8):381. doi: 10.3390/bioengineering9080381.
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 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. -mediated floral dip transformation was used to introduce the IFN-α2a expression cassette into the nuclear genome of red and white rooted 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 L. plants as a safe, cost-effective, and easy-to-use expression system for generating active human IFN-α2a.
人干扰素（IFN）是一种细胞因子，可调节免疫系统对病毒和细菌感染的反应。重组IFN-α已被批准用于治疗多种病毒感染，以及作为治疗各种形式白血病的抗癌药物。本研究的目的是从转基因L.植物中生产功能活性重组人IFN-α2a。因此，构建了一个二元植物表达载体，其包含在花椰菜花叶病毒35SS启动子调控下的IFN-α2a基因编码序列。采用农杆菌介导的花浸泡转化法将IFN-α2a表达盒导入红白根L.植物的核基因组中。从每种基因型中建立了三个独立的转基因株系。检测了部分纯化的重组IFN-α2a蛋白的抗癌和抗病毒活性。分离出的IFN-α2a已被证明可抑制水疱性口炎病毒（VSV）的传播。此外，针对Hep-G2细胞（人肝癌细胞）的细胞毒性和细胞凋亡分析表明，所产生的IFN-α2a作为抗癌剂具有疗效。与细菌、酵母和动物细胞培养系统相比，总体观察结果表明，利用L.植物作为一种安全、经济高效且易于使用的表达系统来生产活性人IFN-α2a是有效的。
