Expression of Heterologous PrP and Prion Propagation in RK13 Cells

Methods Mol Biol. 2017:1658:95-104. doi: 10.1007/978-1-4939-7244-9_8.

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.

Keywords: Cell models; Inducible expression; PrP protein; Prion; Prion titration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / metabolism
  • Brain / pathology
  • Cell Culture Techniques
  • Cell Line
  • Clone Cells
  • Cloning, Molecular
  • Endopeptidase K / chemistry
  • Epithelial Cells / metabolism*
  • Epithelial Cells / pathology
  • Founder Effect*
  • Gene Expression
  • Humans
  • Immunoblotting / methods*
  • Mice
  • Plasmids / chemistry
  • Plasmids / metabolism
  • PrPSc Proteins / chemistry
  • PrPSc Proteins / genetics*
  • PrPSc Proteins / metabolism
  • Protein Folding
  • Rabbits
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • PrPSc Proteins
  • Recombinant Proteins
  • Endopeptidase K