Abstract
Cultured cells are valuable models to study prion infections at the cellular level. Unfortunately, the vast majority of cell lines are resistant to the propagation of prion agents. The rabbit epithelial RK13 cell line is among the few cell lines permissive to prion infection. When genetically engineered to express heterologous PrP proteins, RK13 cells become permissive to several strains of prions from various animal species. Here, we describe the generation of stable RK13 cell clones expressing a heterologous PrP protein in an inducible manner, the establishment and maintenance of chronically infected cultures, and the selection of cell clones suitable for cell-based titration of prions.
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References
Krauss S, Vorberg I (2013) Prions ex vivo: what cell culture models tell us about infectious proteins. Int J Cell Biol 2013:704546
Vilette D (2008) Cell models of prion infection. Vet Res 39:10
Cronier S, Laude H, Peyrin JM (2004) Prions can infect primary cultured neurons and astrocytes and promote neuronal cell death. Proc Natl Acad Sci U S A 101:12271–12276
Bian J, Napier D, Khaychuck V, Angers R, Graham C et al (2010) Cell-based quantification of chronic wasting disease prions. J Virol 84:8322–8326
Arellano-Anaya ZE, Savistchenko J, Mathey J, Huor A, Lacroux C et al (2011) A simple, versatile and sensitive cell-based assay for prions from various species. PLoS One 6:e20563
Klohn PC, Stoltze L, Flechsig E, Enari M, Weissmann C (2003) A quantitative, highly sensitive cell-based infectivity assay for mouse scrapie prions. Proc Natl Acad Sci U S A 100:11666–11671
Neale MH, Mountjoy SJ, Edwards JC, Vilette D, Laude H et al (2010) Infection of cell lines with experimental and natural ovine scrapie agents. J Virol 84:2444–2452
Vilette D, Andreoletti O, Archer F, Madelaine MF, Vilotte JL et al (2001) Ex vivo propagation of infectious sheep scrapie agent in heterologous epithelial cells expressing ovine prion protein. Proc Natl Acad Sci U S A 98:4055–4059
Courageot MP, Daude N, Nonno R, Paquet S, Di Bari MA et al (2008) A cell line infectible by prion strains from different species. J Gen Virol 89:341–347
Lawson VA, Vella LJ, Stewart JD, Sharples RA, Klemm H et al (2008) Mouse-adapted sporadic human Creutzfeldt-Jakob disease prions propagate in cell culture. Int J Biochem Cell Biol 40:2793–2801
Paitel E, Alves da Costa C, Vilette D, Grassi J, Checler F (2002) Overexpression of PrPc triggers caspase 3 activation: potentiation by proteasome inhibitors and blockade by anti-PrP antibodies. J Neurochem 83:1208–1214
Rachidi W, Vilette D, Guiraud P, Arlotto M, Riondel J et al (2003) Expression of prion protein increases cellular copper binding and antioxidant enzyme activities but not copper delivery. J Biol Chem 278:9064–9072
Acknowledgments
This work was partly supported by a grant from ANR-08-MIEN-010 (France) and Z.E. Arellano-Anaya was a recipient of a fellowship from Mexican CONACYT.
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Arellano-Anaya, Z.E., Huor, A., Leblanc, P., Andréoletti, O., Vilette, D. (2017). Expression of Heterologous PrP and Prion Propagation in RK13 Cells. In: Lawson, V. (eds) Prions. Methods in Molecular Biology, vol 1658. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7244-9_8
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DOI: https://doi.org/10.1007/978-1-4939-7244-9_8
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