Efficient recovery of attenuated canine distemper virus from cDNA

Marianne Wyss; Vaiva Gradauskaite; Nadine Ebert; Volker Thiel; Andreas Zurbriggen; Philippe Plattet

doi:10.1016/j.virusres.2022.198796

Efficient recovery of attenuated canine distemper virus from cDNA

Virus Res. 2022 Jul 15:316:198796. doi: 10.1016/j.virusres.2022.198796. Epub 2022 May 11.

Authors

Marianne Wyss¹, Vaiva Gradauskaite², Nadine Ebert³, Volker Thiel³, Andreas Zurbriggen¹, Philippe Plattet⁴

Affiliations

¹ Division of Neurological Sciences, Vetsuisse Faculty, University of Bern, Switzerland.
² Division of Neurological Sciences, Vetsuisse Faculty, University of Bern, Switzerland; Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland.
³ Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, University of Bern, Bern, Switzerland; Institute of Virology and Immunology, Mittelhäusern, Bern, Switzerland.
⁴ Division of Neurological Sciences, Vetsuisse Faculty, University of Bern, Switzerland. Electronic address: philippe.plattet@vetsuisse.unibe.ch.

PMID: 35568090
DOI: 10.1016/j.virusres.2022.198796

Abstract

To provide insights into the biology of the attenuated canine distemper virus (CDV) Onderstepoort (OP) strain (large plaque forming variant), design next-generation multivalent vaccines, or further investigate its promising potential as an oncolytic vector, we employed contemporary modifications to establish an efficient OP-CDV-based reverse genetics platform. Successful viral rescue was obtained however only upon recovery of a completely conserved charged residue (V13E) residing at the N-terminal region of the large protein (L). Although L-V13 and L-V13E did not display drastic differences in cellular localization and physical interaction with P, efficient polymerase complex (P+ L) activity was recorded only with L-V13E. Interestingly, grafting mNeonGreen to the viral N protein via a P2A ribosomal skipping sequence (OP^neon) and its derivative V-protein-knockout variant (OP^neon-V^ko) exhibited delayed replication kinetics in cultured cells. Collectively, we established an efficient OP-CDV-based reverse genetics system that enables the design of various strategies potentially contributing to veterinary medicine and research.

Keywords: Large protein; Morbillivirus; Phosphoprotein; Replication; Reverse genetics; V protein.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line
DNA, Complementary
Distemper Virus, Canine* / genetics
Distemper*
Dogs
Neon
Nucleocapsid Proteins

Substances

DNA, Complementary
Nucleocapsid Proteins
Neon