tetano
Editor, Senior Moderator
Sci Rep. 2019 Nov 4;9(1):15899. doi: 10.1038/s41598-019-52258-7. [h=1]Nanobody-based sandwich reporter system for living cell sensing influenza A virus infection.[/h]
Cao J[SUP]1[/SUP], Zhong N[SUP]2[/SUP], Wang G[SUP]1[/SUP], Wang M[SUP]1[/SUP], Zhang B[SUP]3[/SUP], Fu B[SUP]3[/SUP], Wang Y[SUP]3[/SUP], Zhang T[SUP]1,[/SUP][SUP]3[/SUP], Zhang Y[SUP]1,[/SUP][SUP]3[/SUP], Yang K[SUP]4[/SUP], Chen Y[SUP]5[/SUP], Yuan Q[SUP]6[/SUP], Xia N[SUP]1,[/SUP][SUP]3[/SUP].
[h=3]Author information[/h] 1 State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen, 361102, P.R. China. 2 Concordia International School Shanghai, 345 Huangyang Road Pudong, Shanghai, 201206, P.R. China. 3 State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China. 4 Xiamen International Travel Healthcare Center, Xiamen, China. 5 State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen, 361102, P.R. China. yxchen2008@xmu.edu.cn. 6 State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China. yuanquan@xmu.edu.cn.
[h=3]Abstract[/h] The influenza epidemic is a huge burden to public health. Current influenza vaccines provide limited protection against new variants due to frequent mutation of the virus. The continual emergence of novel variants necessitates the method rapidly monitoring influenza virus infection in experimental systems. Although several replication-competent reporter viruses carrying fluorescent proteins or small luciferase have been generated in previous studies, visualizing influenza virus infection via such strategy requires reverse genetic modification for each viral strain which is usually time-consuming and inconvenient. Here, we created a novel influenza A nucleoprotein (NP) dependent reporter gene transcription activation module using NP-specific nanobodies. Our results demonstrated the modular design allowed reporter genes (mNeonGreen fluorescent protein and Gaussia luciferase) specifically expressing to detect intracellular NP protein, and therefore acts as a universal biosensor to monitor infection of various influenza A subtypes in living cells. The new system may provide a powerful tool to analyze influenza A infections at the cellular level to facilitate new antiviral drug discovery. Moreover, this approach may easily extend to develop live-cell biosensors for other viruses.
PMID: 31685871 DOI: 10.1038/s41598-019-52258-7
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Cao J[SUP]1[/SUP], Zhong N[SUP]2[/SUP], Wang G[SUP]1[/SUP], Wang M[SUP]1[/SUP], Zhang B[SUP]3[/SUP], Fu B[SUP]3[/SUP], Wang Y[SUP]3[/SUP], Zhang T[SUP]1,[/SUP][SUP]3[/SUP], Zhang Y[SUP]1,[/SUP][SUP]3[/SUP], Yang K[SUP]4[/SUP], Chen Y[SUP]5[/SUP], Yuan Q[SUP]6[/SUP], Xia N[SUP]1,[/SUP][SUP]3[/SUP].
[h=3]Author information[/h] 1 State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen, 361102, P.R. China. 2 Concordia International School Shanghai, 345 Huangyang Road Pudong, Shanghai, 201206, P.R. China. 3 State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China. 4 Xiamen International Travel Healthcare Center, Xiamen, China. 5 State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Life Sciences, Xiamen University, Xiamen, 361102, P.R. China. yxchen2008@xmu.edu.cn. 6 State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, School of Public Health, Xiamen University, Xiamen, 361102, China. yuanquan@xmu.edu.cn.
[h=3]Abstract[/h] The influenza epidemic is a huge burden to public health. Current influenza vaccines provide limited protection against new variants due to frequent mutation of the virus. The continual emergence of novel variants necessitates the method rapidly monitoring influenza virus infection in experimental systems. Although several replication-competent reporter viruses carrying fluorescent proteins or small luciferase have been generated in previous studies, visualizing influenza virus infection via such strategy requires reverse genetic modification for each viral strain which is usually time-consuming and inconvenient. Here, we created a novel influenza A nucleoprotein (NP) dependent reporter gene transcription activation module using NP-specific nanobodies. Our results demonstrated the modular design allowed reporter genes (mNeonGreen fluorescent protein and Gaussia luciferase) specifically expressing to detect intracellular NP protein, and therefore acts as a universal biosensor to monitor infection of various influenza A subtypes in living cells. The new system may provide a powerful tool to analyze influenza A infections at the cellular level to facilitate new antiviral drug discovery. Moreover, this approach may easily extend to develop live-cell biosensors for other viruses.
PMID: 31685871 DOI: 10.1038/s41598-019-52258-7
Free full text