miR-281, an abundant midgut-specific miRNA of the vector mosquito Aedes albopictus enhances dengue virus replication
Abstract Background Emerging evidence indicates that microRNAs (miRNAs) are involved in host-virus interaction. We previously reported that some miRNAs were differentially expressed in sugar-fed and blood-fed females of Aedes albopictus (Ae. albopictus). Here, we analysis the role in the host-virus...
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| Format: | Article |
| Language: | English |
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BMC
2014-10-01
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| Series: | Parasites & Vectors |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13071-014-0488-4 |
| _version_ | 1827933607743717376 |
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| author | Yanhe Zhou Yanxia Liu Hui Yan Yiji Li Hao Zhang Jiabao Xu Santhosh Puthiyakunnon Xiaoguang Chen |
| author_facet | Yanhe Zhou Yanxia Liu Hui Yan Yiji Li Hao Zhang Jiabao Xu Santhosh Puthiyakunnon Xiaoguang Chen |
| author_sort | Yanhe Zhou |
| collection | DOAJ |
| description | Abstract Background Emerging evidence indicates that microRNAs (miRNAs) are involved in host-virus interaction. We previously reported that some miRNAs were differentially expressed in sugar-fed and blood-fed females of Aedes albopictus (Ae. albopictus). Here, we analysis the role in the host-virus system of an abundant midgut-specific miRNA in the mosquito Ae. albopictus. Methods The expression profiles of miR-281 in different body parts of Ae. albopictus and following dengue virus infection were determined using RT-qPCR and Northern blot. miR-281 mimics, antagomiRs and corresponding negative controls were designed and their overexpression and knock-down efficiency were analyzed by qRT-PCR after transfecting the mosquito cell lines C6/36, and also by injecting female mosquitoes. Dengue virus serotype-2 (DENV-2) viral genomic RNA abundance was determined by RT-qPCR. The levels of DENV-2 E protein were detected using Western blot. Virus titers were tested using TCID50. RNAhybrid was used to predict targets of miR-281 in the DENV-2 genome. The EGFP plasmid-based reporter system was used to investigate the interaction between miR-281 and the predicted binding site in the C6/36 cell line. Results miR-281 is specifically expressed in the female midgut where dengue virus first invades. After DENV-2 infection, this miRNA is up-regulated in response to viral infection. Functional intervention analyses in vitro with specifically designed miR-281 mimics and corresponding antagomiRs indicated that miR-281 enhances DENV-2 viral replication. Further depletion of miR-281 in female mosquitoes by injection of its specific antagomiRs led to a significant reduction in DENV-2 abundance. The interaction between miR-281 and its predicted target sequence, the DENV-2 genomic 5'-untranslated region (UTR), is confirmed in the context of a plasmid-based reporter system. Conclusion These findings confirm that miR-281, an abundant midgut-specific miRNA, facilitates DENV-2 replication. |
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| id | doaj.art-2a879175cb1e48a99b0bfd0a5fbc87bd |
| institution | Directory Open Access Journal |
| issn | 1756-3305 |
| language | English |
| last_indexed | 2024-03-13T07:28:09Z |
| publishDate | 2014-10-01 |
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| series | Parasites & Vectors |
| spelling | doaj.art-2a879175cb1e48a99b0bfd0a5fbc87bd2023-06-04T11:16:03ZengBMCParasites & Vectors1756-33052014-10-017111110.1186/s13071-014-0488-4miR-281, an abundant midgut-specific miRNA of the vector mosquito Aedes albopictus enhances dengue virus replicationYanhe Zhou0Yanxia Liu1Hui Yan2Yiji Li3Hao Zhang4Jiabao Xu5Santhosh Puthiyakunnon6Xiaoguang Chen7Key Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical UniversityKey Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical UniversityKey Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical UniversityKey Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical UniversityKey Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical UniversityKey Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical UniversityKey Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical UniversityKey Laboratory of Prevention and Control for Emerging Infectious Diseases of Guangdong, Department of Pathogen Biology, School of Public Health and Tropical Medicine, Southern Medical UniversityAbstract Background Emerging evidence indicates that microRNAs (miRNAs) are involved in host-virus interaction. We previously reported that some miRNAs were differentially expressed in sugar-fed and blood-fed females of Aedes albopictus (Ae. albopictus). Here, we analysis the role in the host-virus system of an abundant midgut-specific miRNA in the mosquito Ae. albopictus. Methods The expression profiles of miR-281 in different body parts of Ae. albopictus and following dengue virus infection were determined using RT-qPCR and Northern blot. miR-281 mimics, antagomiRs and corresponding negative controls were designed and their overexpression and knock-down efficiency were analyzed by qRT-PCR after transfecting the mosquito cell lines C6/36, and also by injecting female mosquitoes. Dengue virus serotype-2 (DENV-2) viral genomic RNA abundance was determined by RT-qPCR. The levels of DENV-2 E protein were detected using Western blot. Virus titers were tested using TCID50. RNAhybrid was used to predict targets of miR-281 in the DENV-2 genome. The EGFP plasmid-based reporter system was used to investigate the interaction between miR-281 and the predicted binding site in the C6/36 cell line. Results miR-281 is specifically expressed in the female midgut where dengue virus first invades. After DENV-2 infection, this miRNA is up-regulated in response to viral infection. Functional intervention analyses in vitro with specifically designed miR-281 mimics and corresponding antagomiRs indicated that miR-281 enhances DENV-2 viral replication. Further depletion of miR-281 in female mosquitoes by injection of its specific antagomiRs led to a significant reduction in DENV-2 abundance. The interaction between miR-281 and its predicted target sequence, the DENV-2 genomic 5'-untranslated region (UTR), is confirmed in the context of a plasmid-based reporter system. Conclusion These findings confirm that miR-281, an abundant midgut-specific miRNA, facilitates DENV-2 replication.https://doi.org/10.1186/s13071-014-0488-4Aedes albopictusmiRNADengue virusPathogen-host interaction |
| spellingShingle | Yanhe Zhou Yanxia Liu Hui Yan Yiji Li Hao Zhang Jiabao Xu Santhosh Puthiyakunnon Xiaoguang Chen miR-281, an abundant midgut-specific miRNA of the vector mosquito Aedes albopictus enhances dengue virus replication Parasites & Vectors Aedes albopictus miRNA Dengue virus Pathogen-host interaction |
| title | miR-281, an abundant midgut-specific miRNA of the vector mosquito Aedes albopictus enhances dengue virus replication |
| title_full | miR-281, an abundant midgut-specific miRNA of the vector mosquito Aedes albopictus enhances dengue virus replication |
| title_fullStr | miR-281, an abundant midgut-specific miRNA of the vector mosquito Aedes albopictus enhances dengue virus replication |
| title_full_unstemmed | miR-281, an abundant midgut-specific miRNA of the vector mosquito Aedes albopictus enhances dengue virus replication |
| title_short | miR-281, an abundant midgut-specific miRNA of the vector mosquito Aedes albopictus enhances dengue virus replication |
| title_sort | mir 281 an abundant midgut specific mirna of the vector mosquito aedes albopictus enhances dengue virus replication |
| topic | Aedes albopictus miRNA Dengue virus Pathogen-host interaction |
| url | https://doi.org/10.1186/s13071-014-0488-4 |
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