Jingjing Liu# , Fang Wang# , Jiangang Zhao , Yinglin Qi , Jitao Chang , Chao Sun , Zhigang Jiang , Junwei Ge , Xin Yin

Front Microbiol.2024 Nov 14:15:1458771.doi: 10.3389/fmicb.2024.1458771. eCollection 2024.

Abstract

Introduction: Akabane virus (AKAV) is a worldwide epidemic arbovirus belonging to the Bunyavirales order that predominantly infects livestock and causes severe congenital malformations. Reporter-expressing recombinant virus represents a powerful tool to characterize the viral biology  in vitro  and  in vivo .

Methods: In this study, we have successfully established a reverse genetics system for AKAV. The recued virus possessed similar growth characteristics to the parental virus  in vitro . Moreover, the recombinant AKAV reporter viruses expressing nanoluciferase (Nluc) or mWasabi were constructed by inserting into S segment, named rAKAV-Nluc and rAKAV-mWasabi, respectively.

Results: We investigated the virological characteristics of rAKAV-Nluc and rAKAV-mWasabi and found that rAKAV-Nluc displayed similar growth kinetics as the parental virus and could stably produce the nano-luciferase even after 10 rounds of serial passages. rAKAV-mWasabi also exhibited comparable growth kinetics and genetic stability as the parental virus. We further used the two reporter viruses to test the susceptibility of different cell lines to AKAV and found that cell lines derived from various host species, including human, swine, cattle, and monkey enables AKAV replication efficiently, accelerating our understanding of the AKAV cell tropism range.

Discussion: Taken together, our established reverse genetics system for AKAV provides more convenient screening tools and can be used to study AKAV virulence and tropism, and to elucidate the molecular biology of AKAV.

Keywords: Akabane virus; cell tropism; mWasabi; nanoluciferase; reporter-expressing virus; reverse genetics system.