Generation of a GFP reporter Akabane virus with enhanced fluorescence intensity by modification of artificial ambisense S genome
Akiko Takenaka-Uema, Shin Murakami, Nanako Ushio, Tomoya Kobayashi-Kitamura, Masashi Uema, Kazuyuki Uchida, Taisuke Horimoto
We previously generated a recombinant reporter Akabane virus expressing enhanced green fluorescence protein (eGFP-AKAV), with an artificial S genome encoding eGFP in the ambisense RNA. Although the eGFP-AKAV was able to detect infected cells in in vivo histopathological study, its fluorescent signal was too weak to apply to in vivo imaging study. Here, we successfully generated a modified reporter, eGFP/38-AKAV, with 38-nucleotide deletion of the internal region of the 5′ untranslated region of S RNA. The eGFP/38-AKAV expressed higher intensity of eGFP fluorescence both in vitro and in vivo than the original eGFP-AKAV did. In addition, eGFP/38-AKAV was pathogenic in mice at a comparable level to that in wild-type AKAV. In the mice infected with eGFP/38-AKAV, the fluorescent signals, i.e., the virus-infected cells, were detected in the central nervous system using the whole-organ imaging. Our findings indicate that eGFP/38-AKAV could be used as a powerful tool to help elucidate the dynamics of AKAV in vivo.
- : Viruses
- : 10.3390/v11070634
- : https://www.mdpi.com/1999-4915/11/7/634/htm