Author: Hölzer, Martin; Schoen, Andreas; Wulle, Julia; Müller, Marcel A.; Drosten, Christian; Marz, Manja; Weber, Friedemann
Title: Virus- and Interferon Alpha-Induced Transcriptomes of Cells from the Microbat Myotis daubentonii Document date: 2019_8_10
ID: 0co6m9af_9
Snippet: To elucidate the overall antiviral potency and the unimpeded ability to produce IFN in response to infection, we employed a mutant of Rift Valley fever virus that has its IFN suppressor NSs replaced by the Renilla luciferase gene (RVFVDNSs::Renilla) (Kuri et al., 2010) . RVFVDNSs::Renilla is a strong inducer of type I IFNs, is sensitive to the antiviral action of IFNs, and able to infect cells from a wide range of vertebrates (Kuri et al., 2010) .....
Document: To elucidate the overall antiviral potency and the unimpeded ability to produce IFN in response to infection, we employed a mutant of Rift Valley fever virus that has its IFN suppressor NSs replaced by the Renilla luciferase gene (RVFVDNSs::Renilla) (Kuri et al., 2010) . RVFVDNSs::Renilla is a strong inducer of type I IFNs, is sensitive to the antiviral action of IFNs, and able to infect cells from a wide range of vertebrates (Kuri et al., 2010) . The built-in Renilla luciferase facilitates rapid and quantitative measurements of viral replication. Also, the M. daubentonii MyDauNi/2c cells could be infected with our reporter virus, permitting replication levels similar as in human A549 lung cell line or the fetal R. aegytiacus line Ro6E-J (Jordan et al., 2009) (Figure S1B) . Moreover, treatment with the IFN signaling inhibitor Ruxolitinib increased RVFVDNSs::Renilla replication, whereas IFN-a suppressed it. Although the cell lines are from different organs, these results, taken together, demonstrate that our immortalized Yangochiroptera/Vespertilionidae cell line MyDauNi/2c possesses a fully functional IFN system that is comparable with the one of human and megabat cell lines. Figure 1A gives an overview of the experimental setup. The MyDauNi/2c cells were either mock treated, infected with a strong IFN inducer (RVFV NSs mutant Clone 13) , or treated with 1,000 U/ml IFN-a. Total cell RNAs were isolated at 6 and 24 h post infection, and rRNA-depleted cDNA libraries were sequenced with 60-70 million strand-specific single-end reads per sample (Table S1 ). Of note, all experiments were performed in three independent, biological replicates.
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