A CRISPR/Cas9 Generated Bovine CD46-knockout Cell Line—A Tool to Elucidate the Adaptability of Bovine Viral Diarrhea Viruses (BVDV).

Kevin P. Szillat , Susanne Koethe, Kerstin Wernike , Dirk Höper and Martin Beer

Cellular CD46 is a type 1 transmembrane glycoprotein expressed on all nucleated cells that can serve as a binding partner for pathogens like certain adenoviruses, bovine viral diarrhea virus (BVDV) and classical swine fever virus. To study adaptation mechanisms of pestiviruses, a CD46-negative cellular environment was created using CRISPR/Cas9 and Madin–Darby Bovine Kidney cells. In the absence of CD46, pestivirus A and B species (BVDV-1 and -2) could still enter the cell, although it was only a fraction of the virus particles. Continuous passaging enriched for mutant viruses with crucial compensatory amino acid substitutions in the viral ERNS glycoprotein.

Published, August 2020, in Viruses, in the Special Issue Bovine Viral Diarrhea Virus and Related Pestiviruses

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Host switching pathogens, infectious outbreaks and zoonosis; a Marie Sklodowska-Curie Innovative Training Network.

This project has received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement No 721367.