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

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.