Arellano-Anaya Zaira E, Huor Alvina, Leblanc Pascal, Andréoletti Olivier, Vilette Didier
INRA, UMR 1225, IHAP, 31076, Toulouse, France.
Université de Toulouse, INP, ENVT, UMR1225, IHAP, 31076, Toulouse, France.
Methods Mol Biol. 2017;1658:95-104. doi: 10.1007/978-1-4939-7244-9_8.
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.
培养细胞是在细胞水平研究朊病毒感染的有价值模型。不幸的是，绝大多数细胞系对朊病毒因子的增殖具有抗性。兔上皮RK13细胞系是少数允许朊病毒感染的细胞系之一。当通过基因工程改造以表达异源PrP蛋白时，RK13细胞对来自各种动物物种的几种朊病毒株变得敏感。在这里，我们描述了以诱导方式表达异源PrP蛋白的稳定RK13细胞克隆的产生、慢性感染培养物的建立和维持，以及适合基于细胞的朊病毒滴定的细胞克隆的选择。
