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.


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 (OPneon) and its derivative V-protein-knockout variant (OPneon-Vko) 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


  • DNA, Complementary
  • Nucleocapsid Proteins
  • Neon