Construction of HiBiT-tagged porcine deltacoronavirus via NanoBiT split-luciferase system and its utility in antiviral research.

Abstract

Porcine deltacoronavirus (PDCoV) is a novel coronavirus that causes acute diarrhea and high mortality in piglets, and poses a significant threat to public health due to its potential for cross-species transmission. HiBiT is a tag that can bind to LgBiT to restore NanoBiT activity, and it can be inserted into viral genomes to generate reporter viruses. In this study We successfully constructed an infectious clone of PDCoV using the transformation-associated recombination (TAR) technique in yeast. The rescued virus exhibited biological characteristics similar to those of the wild-type strain. Based on this infectious clone, we insert a HiBiT tag into the NS6 gene and successfully rescued the recombinant virus, which also showed comparable biological properties to the wild-type virus. The functionality of the HiBiT reporter gene was validated using two antiviral compounds and compared with an eGFP-tagged reporter virus. The results demonstrated that the PDCoV reporter virus carrying the HiBiT tag provided stronger peak signals, higher sensitivity, and better genetic stability for antiviral drug screening These findings suggest that the HiBiT-tagged PDCoV is a robust tool for antiviral drug discovery and can facilitate the development of therapeutics against PDCoV.