Bioluminescent and fluorescent reporter-expressing recombinant Akabane virus (AKAV): an excellent tool to dissect viral replication
IntroductionAkabane virus (AKAV) is a worldwide epidemic arbovirus belonging to the Bunyavirales order that predominantly infects livestock and causes severe congenital malformations. Reporter-expressing recombinant virus represents a powerful tool to characterize the viral biology in vitro and in v...
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Frontiers Media S.A.
2024-11-01
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| author | Jingjing Liu Jingjing Liu Fang Wang Jiangang Zhao Jiangang Zhao Yinglin Qi Jitao Chang Jitao Chang Chao Sun Zhigang Jiang Junwei Ge Xin Yin |
| author_facet | Jingjing Liu Jingjing Liu Fang Wang Jiangang Zhao Jiangang Zhao Yinglin Qi Jitao Chang Jitao Chang Chao Sun Zhigang Jiang Junwei Ge Xin Yin |
| author_sort | Jingjing Liu |
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| description | IntroductionAkabane virus (AKAV) is a worldwide epidemic arbovirus belonging to the Bunyavirales order that predominantly infects livestock and causes severe congenital malformations. Reporter-expressing recombinant virus represents a powerful tool to characterize the viral biology in vitro and in vivo.MethodsIn this study, we have successfully established a reverse genetics system for AKAV. The recued virus possessed similar growth characteristics to the parental virus in vitro. Moreover, the recombinant AKAV reporter viruses expressing nanoluciferase (Nluc) or mWasabi were constructed by inserting into S segment, named rAKAV-Nluc and rAKAV-mWasabi, respectively.ResultsWe investigated the virological characteristics of rAKAV-Nluc and rAKAV-mWasabi and found that rAKAV-Nluc displayed similar growth kinetics as the parental virus and could stably produce the nano-luciferase even after 10 rounds of serial passages. rAKAV-mWasabi also exhibited comparable growth kinetics and genetic stability as the parental virus. We further used the two reporter viruses to test the susceptibility of different cell lines to AKAV and found that cell lines derived from various host species, including human, swine, cattle, and monkey enables AKAV replication efficiently, accelerating our understanding of the AKAV cell tropism range.DiscussionTaken together, our established reverse genetics system for AKAV provides more convenient screening tools and can be used to study AKAV virulence and tropism, and to elucidate the molecular biology of AKAV. |
| format | Article |
| id | doaj-art-4fee0fdfaba24cac8ca527830ae9016b |
| institution | OA Journals |
| issn | 1664-302X |
| language | English |
| publishDate | 2024-11-01 |
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| series | Frontiers in Microbiology |
| spelling | doaj-art-4fee0fdfaba24cac8ca527830ae9016b2025-08-20T02:14:57ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2024-11-011510.3389/fmicb.2024.14587711458771Bioluminescent and fluorescent reporter-expressing recombinant Akabane virus (AKAV): an excellent tool to dissect viral replicationJingjing Liu0Jingjing Liu1Fang Wang2Jiangang Zhao3Jiangang Zhao4Yinglin Qi5Jitao Chang6Jitao Chang7Chao Sun8Zhigang Jiang9Junwei Ge10Xin Yin11State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaCollege of Veterinary Medicine, Northeast Agricultural University, Harbin, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaCollege of Animal Science and Technology, Tarim University, Alar, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaInstitute of Western Agriculture, The Chinese Academy of Agricultural Sciences, Changji, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaCollege of Veterinary Medicine, Northeast Agricultural University, Harbin, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaIntroductionAkabane virus (AKAV) is a worldwide epidemic arbovirus belonging to the Bunyavirales order that predominantly infects livestock and causes severe congenital malformations. Reporter-expressing recombinant virus represents a powerful tool to characterize the viral biology in vitro and in vivo.MethodsIn this study, we have successfully established a reverse genetics system for AKAV. The recued virus possessed similar growth characteristics to the parental virus in vitro. Moreover, the recombinant AKAV reporter viruses expressing nanoluciferase (Nluc) or mWasabi were constructed by inserting into S segment, named rAKAV-Nluc and rAKAV-mWasabi, respectively.ResultsWe investigated the virological characteristics of rAKAV-Nluc and rAKAV-mWasabi and found that rAKAV-Nluc displayed similar growth kinetics as the parental virus and could stably produce the nano-luciferase even after 10 rounds of serial passages. rAKAV-mWasabi also exhibited comparable growth kinetics and genetic stability as the parental virus. We further used the two reporter viruses to test the susceptibility of different cell lines to AKAV and found that cell lines derived from various host species, including human, swine, cattle, and monkey enables AKAV replication efficiently, accelerating our understanding of the AKAV cell tropism range.DiscussionTaken together, our established reverse genetics system for AKAV provides more convenient screening tools and can be used to study AKAV virulence and tropism, and to elucidate the molecular biology of AKAV.https://www.frontiersin.org/articles/10.3389/fmicb.2024.1458771/fullAkabane virusreverse genetics systemreporter-expressing virusnanoluciferasemWasabicell tropism |
| spellingShingle | Jingjing Liu Jingjing Liu Fang Wang Jiangang Zhao Jiangang Zhao Yinglin Qi Jitao Chang Jitao Chang Chao Sun Zhigang Jiang Junwei Ge Xin Yin Bioluminescent and fluorescent reporter-expressing recombinant Akabane virus (AKAV): an excellent tool to dissect viral replication Frontiers in Microbiology Akabane virus reverse genetics system reporter-expressing virus nanoluciferase mWasabi cell tropism |
| title | Bioluminescent and fluorescent reporter-expressing recombinant Akabane virus (AKAV): an excellent tool to dissect viral replication |
| title_full | Bioluminescent and fluorescent reporter-expressing recombinant Akabane virus (AKAV): an excellent tool to dissect viral replication |
| title_fullStr | Bioluminescent and fluorescent reporter-expressing recombinant Akabane virus (AKAV): an excellent tool to dissect viral replication |
| title_full_unstemmed | Bioluminescent and fluorescent reporter-expressing recombinant Akabane virus (AKAV): an excellent tool to dissect viral replication |
| title_short | Bioluminescent and fluorescent reporter-expressing recombinant Akabane virus (AKAV): an excellent tool to dissect viral replication |
| title_sort | bioluminescent and fluorescent reporter expressing recombinant akabane virus akav an excellent tool to dissect viral replication |
| topic | Akabane virus reverse genetics system reporter-expressing virus nanoluciferase mWasabi cell tropism |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2024.1458771/full |
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