Construction of a Dengue NanoLuc Reporter Virus for In Vivo Live Imaging in Mice |
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| Authors: | Enyue Fang Xiaohui Liu Miao Li Jingjing Liu Zelun Zhang Xinyu Liu Xingxing Li Wenjuan Li Qinhua Peng Yongxin Yu Yuhua Li |
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| Affiliation: | 1.Department of Arbovirus Vaccine, National Institutes for Food and Drug Control, Beijing 102629, China; (E.F.); (X.L.); (M.L.); (J.L.); (Z.Z.); (X.L.); (X.L.); (W.L.); (Q.P.);2.Wuhan Institute of Biological Products, Co., Ltd., Wuhan 430207, China |
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| Abstract: | Since the first isolation in 1943, the dengue virus (DENV) has spread throughout the world, but effective antiviral drugs or vaccines are still not available. To provide a more stable reporter DENV for vaccine development and antiviral drug screening, we constructed a reporter DENV containing the NanoLuc reporter gene, which was inserted into the 5′ untranslated region and capsid junction region, enabling rapid virus rescue by in vitro ligation. In addition, we established a live imaging mouse model and found that the reporter virus maintained the neurovirulence of prototype DENV before engineering. DENV-4 exhibited dramatically increased neurovirulence following a glycosylation site-defective mutation in the envelope protein. Significant mice mortality with neurological onset symptoms was observed after intracranial infection of wild-type (WT) mice, thus providing a visualization tool for DENV virulence assessment. Using this model, DENV was detected in the intestinal tissues of WT mice after infection, suggesting that intestinal lymphoid tissues play an essential role in DENV pathogenesis. |
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| Keywords: | flavivirus reporter virus in vivo imaging neurovirulence |
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