Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection
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 gener...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2019-10-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2095311919627176 |
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| author | Dan-dan ZHAO Lei SHUAI Jin-ying GE Jin-liang WANG Zhi-yuan WEN Ren-qiang LIU Chong WANG Xi-jun WANG Zhi-gao BU |
| author_facet | Dan-dan ZHAO Lei SHUAI Jin-ying GE Jin-liang WANG Zhi-yuan WEN Ren-qiang LIU Chong WANG Xi-jun WANG Zhi-gao BU |
| author_sort | Dan-dan ZHAO |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-55cd8c4af5f64d2eb59ec0a0fe21690b |
| institution | Kabale University |
| issn | 2095-3119 |
| language | English |
| publishDate | 2019-10-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj-art-55cd8c4af5f64d2eb59ec0a0fe21690b2025-08-20T03:56:12ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192019-10-0118102361236810.1016/S2095-3119(19)62717-6Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infectionDan-dan ZHAO0Lei SHUAI1Jin-ying GE2Jin-liang WANG3Zhi-yuan WEN4Ren-qiang LIU5Chong WANG6Xi-jun WANG7Zhi-gao BU8State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.ChinaState Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, P.R.China; Correspondence BU Zhi-gaoThe 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.http://www.sciencedirect.com/science/article/pii/S2095311919627176rabies virusself-fluorescencebindinginternalization |
| spellingShingle | Dan-dan ZHAO Lei SHUAI Jin-ying GE Jin-liang WANG Zhi-yuan WEN Ren-qiang LIU Chong WANG Xi-jun WANG Zhi-gao BU Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection Journal of Integrative Agriculture rabies virus self-fluorescence binding internalization |
| title | Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection |
| title_full | Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection |
| title_fullStr | Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection |
| title_full_unstemmed | Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection |
| title_short | Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection |
| title_sort | generation of recombinant rabies virus era strain applied to virus tracking in cell infection |
| topic | rabies virus self-fluorescence binding internalization |
| url | http://www.sciencedirect.com/science/article/pii/S2095311919627176 |
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