Cloning and expression of an anti-cancerous cytokine: human IL-29 gene in Chlamydomonas reinhardtii

Abstract

Green algae, Chlamydomonas reinhardtii, with low cultivation cost, absence of endotoxins and insusceptibility to human pathogens is emerging as a potential system for the future production of recombinant proteins. The recent development of molecular tools enabling recombinant protein expression in algae chloroplast has provided new research and advance opportunities for developing low-cost therapeutic proteins. In the present study, algae chloroplast expression system was evaluated for the recombinant production of an anti-cancerous therapeutic protein, Interleukin 29 (IL29). The IL29 gene was cloned into algae chloroplast expression vector (pSRSapI). After the transformation, the positive clones were screened for homoplasmy and the presence of the IL29 gene by spot test and PCR analysis, respectively. The expressed SDS-PAGE and western blotting assay characterized IL-29. The algae expressed IL-29 was biologically active in an anti-proliferating bioassay using HepG2 cells. The results suggest that the Chlamydomonas reinhardtii expression system is convenient, low-cost, eco-friendly, and safe to express IL29.

Enhanced soluble expression of active recombinant human interleukin-29 using champion pET SUMO system

“…The yield in current study was higher (98 mg/l) in comparison to other studies who have reported as 60 mg/l (Li & He 2006 ), 65 mg/l (Xie et al 2007 ), and 70 mg/l (Shaldzhyan et al 2021 ). The antiproliferation activity of rhIL-29 in HepG2, HCT-116, and MCF-7 cell lines was consistent with previous reports (Fujie et al 2011 ; Balabanov et al 2019 ; Akram et al 2023 ; Rahman et al 2023 ), indicating that our research is successful in producing functional IL-29.…”

Accelerating Chloroplast Engineering: A New System for Rapid Generation of Marker-Free Transplastomic Lines of Chlamydomonas reinhardtii

“…Furthermore, the deleted plastome region was designed to extend as far as a well-established neutral locus for transgene insertion, ensuring that psbH restoration was always accompanied by transgene insertion. TN72 has been used by several groups to create marker-free transgenic lines producing recombinant therapeutics [ 14 , 15 , 16 , 17 , 18 , 19 ]. However, TN72 has several drawbacks as a platform strain.…”

Functional and Extracellular Production and Antitumor Activity of Mouse Alpha Klotho in Model Microalga Chlamydomonas reinhardtii

“…Microalgae stands as a promising biological platform for the cost‐effective and sustainable production of recombinant proteins and bioproducts using only water, CO 2, and sunlight. The microalgae C. reinhardtii is one of the most studied algae species and has been developed as a potent and photosynthetic expression system for the production of recombinant proteins, antigens, fatty acids, and chemicals with wide industrial applications [21–24] . In addition, C. reinhardtii can be grown autotrophically, heterotrophically, or even mixotrophically using various carbon sources [25] .…”

“…The microalgae C. reinhardtii is one of the most studied algae species and has been developed as a potent and photosynthetic expression system for the production of recombinant proteins, antigens, fatty acids, and chemicals with wide industrial applications. [21][22][23][24] In addition, C. reinhardtii can be grown autotrophically, heterotrophically, or even mixotrophically using various carbon sources. [25] Well-documented molecular tools serving the efficient and high-level expression of transgenes in nuclear and chloroplast genomes have also been developed, making microalgae suitable hosts for synthetic biology and biotechnology.…”