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

Overview

Journal Mol Biotechnol

Publisher Springer

Specialties Biotechnology
Molecular Biology

Date 2018 Dec 14

PMID 30543053

Citations 11

Authors

Yassin El-Ayouty

Islam El-Manawy

Sherif Nasih

Emad Hamdy

Rashad Kebeish

Affiliations

Soon will be listed here.

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.

Citing Articles

Towards chlorocytes for therapeutic intravascular photosynthesis.

Vargas-Torres V, Becerra D, Boric M, Egana J Appl Microbiol Biotechnol. 2024; 108(1):489.

PMID: 39417888 PMC: 11486813. DOI: 10.1007/s00253-024-13285-1.

Harnessing the potential of microalgae for the production of monoclonal antibodies and other recombinant proteins.

Rajput B, Ikram S, Tripathi B Protoplasma. 2024; 261(6):1105-1125.

PMID: 38970700 DOI: 10.1007/s00709-024-01967-6.

The Clinical Promise of Microalgae in Rheumatoid Arthritis: From Natural Compounds to Recombinant Therapeutics.

Cutolo E, Caferri R, Campitiello R, Cutolo M Mar Drugs. 2023; 21(12).

PMID: 38132951 PMC: 10745133. DOI: 10.3390/md21120630.

Current Nuclear Engineering Strategies in the Green Microalga .

Perozeni F, Baier T Life (Basel). 2023; 13(7).

PMID: 37511941 PMC: 10381326. DOI: 10.3390/life13071566.

Comparing the Ability of Secretory Signal Peptides for Heterologous Expression of Anti-Lipopolysaccharide Factor 3 in .

Zhuang H, Ou Y, Chen R, Huang D, Wang C Mar Drugs. 2023; 21(6).

PMID: 37367671 PMC: 10301575. DOI: 10.3390/md21060346.

References

Llovet J, Sala M, Castells L, Suarez Y, Vilana R, Bianchi L . Randomized controlled trial of interferon treatment for advanced hepatocellular carcinoma. Hepatology. 1999; 31(1):54-8. DOI: 10.1002/hep.510310111. View

Frank C, Mohamed M, Strickland G, Lavanchy D, Arthur R, Magder L . The role of parenteral antischistosomal therapy in the spread of hepatitis C virus in Egypt. Lancet. 2001; 355(9207):887-91. DOI: 10.1016/s0140-6736(99)06527-7. View

Melen K, Keskinen P, Lehtonen A, Julkunen I . Interferon-induced gene expression and signaling in human hepatoma cell lines. J Hepatol. 2000; 33(5):764-72. DOI: 10.1016/s0168-8278(00)80308-6. View

Gamero A, Larner A . Vanadate facilitates interferon alpha-mediated apoptosis that is dependent on the Jak/Stat pathway. J Biol Chem. 2001; 276(17):13547-53. DOI: 10.1074/jbc.M007948200. View

Harris E . CHLAMYDOMONAS AS A MODEL ORGANISM. Annu Rev Plant Physiol Plant Mol Biol. 2001; 52:363-406. DOI: 10.1146/annurev.arplant.52.1.363. View

Shepard C, Finelli L, Alter M . Global epidemiology of hepatitis C virus infection. Lancet Infect Dis. 2005; 5(9):558-67. DOI: 10.1016/S1473-3099(05)70216-4. View

Elshal M, McCoy J . Multiplex bead array assays: performance evaluation and comparison of sensitivity to ELISA. Methods. 2006; 38(4):317-23. PMC: 1534009. DOI: 10.1016/j.ymeth.2005.11.010. View

Lansky E, Newman R . Punica granatum (pomegranate) and its potential for prevention and treatment of inflammation and cancer. J Ethnopharmacol. 2006; 109(2):177-206. DOI: 10.1016/j.jep.2006.09.006. View

Griesbeck C, Kobl I, Heitzer M . Chlamydomonas reinhardtii: a protein expression system for pharmaceutical and biotechnological proteins. Mol Biotechnol. 2006; 34(2):213-23. DOI: 10.1385/MB:34:2:213. View

10.

Abdel-Hamid M, El-Daly M, Molnegren V, El-Kafrawy S, Abdel-Latif S, Esmat G . Genetic diversity in hepatitis C virus in Egypt and possible association with hepatocellular carcinoma. J Gen Virol. 2007; 88(Pt 5):1526-1531. DOI: 10.1099/vir.0.82626-0. View

11.

Scarzello A, Romero-Weaver A, Maher S, Veenstra T, Zhou M, Qin A . A Mutation in the SH2 domain of STAT2 prolongs tyrosine phosphorylation of STAT1 and promotes type I IFN-induced apoptosis. Mol Biol Cell. 2007; 18(7):2455-62. PMC: 1924825. DOI: 10.1091/mbc.e06-09-0843. View

12.

Zhou Z, Hamming O, Ank N, Paludan S, Nielsen A, Hartmann R . Type III interferon (IFN) induces a type I IFN-like response in a restricted subset of cells through signaling pathways involving both the Jak-STAT pathway and the mitogen-activated protein kinases. J Virol. 2007; 81(14):7749-58. PMC: 1933366. DOI: 10.1128/JVI.02438-06. View

13.

Niessen M, Thiruveedhi K, Rosenkranz R, Kebeish R, Hirsch H, Kreuzaler F . Mitochondrial glycolate oxidation contributes to photorespiration in higher plants. J Exp Bot. 2007; 58(10):2709-15. DOI: 10.1093/jxb/erm131. View

14.

Hagiwara S, Kudo M, Nakatani T, Sakaguchi Y, Nagashima M, Fukuta N . Combination therapy with PEG-IFN-alpha and 5-FU inhibits HepG2 tumour cell growth in nude mice by apoptosis of p53. Br J Cancer. 2007; 97(11):1532-7. PMC: 2360253. DOI: 10.1038/sj.bjc.6604058. View

15.

Antonelli G . Biological basis for a proper clinical application of alpha interferons. New Microbiol. 2008; 31(3):305-18. View

16.

Liu C, Chuang W, Lee C, Yu M, Lu S, Wu S . Peginterferon alfa-2a plus ribavirin for the treatment of dual chronic infection with hepatitis B and C viruses. Gastroenterology. 2008; 136(2):496-504.e3. DOI: 10.1053/j.gastro.2008.10.049. View

17.

Trinchieri G . Type I interferon: friend or foe?. J Exp Med. 2010; 207(10):2053-63. PMC: 2947062. DOI: 10.1084/jem.20101664. View

18.

Obembe O, Popoola J, Leelavathi S, Reddy S . Advances in plant molecular farming. Biotechnol Adv. 2010; 29(2):210-22. DOI: 10.1016/j.biotechadv.2010.11.004. View

19.

Rosales-Mendoza S, Paz-Maldonado L, Soria-Guerra R . Chlamydomonas reinhardtii as a viable platform for the production of recombinant proteins: current status and perspectives. Plant Cell Rep. 2011; 31(3):479-94. DOI: 10.1007/s00299-011-1186-8. View

20.

Li H, Liu Q, Cui K, Liu J, Ren Y, Shi D . Expression of biologically active human interferon alpha 2b in the milk of transgenic mice. Transgenic Res. 2012; 22(1):169-78. DOI: 10.1007/s11248-012-9623-1. View