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Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection

作者：

Zhao Dan-dan;Shuai Lei;Ge Jin-ying;Wang Jin-liang;Wen Zhi-yuan;Liu Ren-qiang;Wang Chong;Wang Xi-jun;Bu Zhi-gao

单位：

Yangzhou Univ, Jiangsu Coinnovat Ctr Prevent & Control Importan, Yangzhou 225009, Jiangsu, Peoples R China;Chinese Acad Agr Sci, State Key Lab Vet Biotechnol, Harbin Vet Res Inst, Harbin 150069, Heilongjiang, Peoples R China

关键词：

rabies virus;self-fluorescence;binding;internalization

摘要：

The mechanism of rabies virus (RABV) infection still needs to be further characterized. RABV particle with self-fluorescent is a powerful viral model to visualize the viral infection process in cells. Herein, based on a reverse genetic system of the Evelyn-Rokitnicki-Abelseth (rERA) strain, we generated a recombinant RABV rERA-N/mCherry strain that stably expresses an additional ERA nucleoprotein that fuses with the red fluorescent protein mCherry (N/mCherry). The rERA-N/mCherry strain retained growth property similar to the parent strain rERA in vitro. The N/mCherry expression showed genetic stability during passage into mouse neuroblastoma (NA) cells and did not change the virulence of the vector. The rERA-N/mCherry strain was then utilized as a visual viral model to study the RABV-cell binding and internalization. We directly observed the red self-fluorescence of rERA-N/mCherry particles binding to the cell surface, and further co-localizing with clathrin in the early stage of infection in NA cells by fluorescence microscopy. Our results showed that the rERA-N/mCherry strain uses clathrin-dependent endocytosis to enter cells, which is consistent with the well-known mechanism of RABV invasion. The recombinant RABV rERA-N/mCherry thus appears to have the potential to be an effective viral model to further explore the fundamental molecular mechanism of rabies neuropathogenesis.

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Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection

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