Development of a reporter HBoV1 strain for antiviral drug screening and life cycle studies

Jielin Tang; Sijie Chen; Yi Zhong; Yijun Deng; Dan Huang; Junjun Liu; Yi Zheng; Jiyuan Xu; Bao Xue; Fan Wang; Yuan Zhou; Hanzhong Wang; Qi Yang; Xinwen Chen

doi:10.1016/j.virs.2025.03.009

April 2025

Citation: Jielin Tang, Sijie Chen, Yi Zhong, Yijun Deng, Dan Huang, Junjun Liu, Yi Zheng, Jiyuan Xu, Bao Xue, Fan Wang, Yuan Zhou, Hanzhong Wang, Qi Yang, Xinwen Chen. Development of a reporter HBoV1 strain for antiviral drug screening and life cycle studies .VIROLOGICA SINICA, 2025, 40(2) : 275-283. http://dx.doi.org/10.1016/j.virs.2025.03.009

Development of a reporter HBoV1 strain for antiviral drug screening and life cycle studies

Jielin Tang ^{a,b
,,} ,
Sijie Chen ^a,b ,
Yi Zhong ^a ,
Yijun Deng ^a,b ,
Dan Huang ^a,c ,
Junjun Liu ^a ,
Yi Zheng ^c ,
Jiyuan Xu ^a,c ,
Bao Xue ^a,c ,
Fan Wang ^d ,
Yuan Zhou ^a ,
Hanzhong Wang ^c ,
Qi Yang ^{a,b
,,} ,
Xinwen Chen ^a,b

a. Guangzhou National Laboratory, Guangzhou 510005, China;
b. State Key Laboratory of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou 511436, China;
c. State Key Laboratory of Virology, Wuhan Institute of Virology, Center for Biosafety Mega-Science, Chinese Academy of Sciences, Wuhan 430071, China;
d. GMU-GIBH Joint School of Life Sciences, Guangzhou Medical University, Guangzhou 511436, China

Corresponding author: Jielin Tang, tang_jielin@gzlab.ac.cn
Qi Yang, yang_qi@gzlab.ac.cn
Received Date: 30 September 2024
Accepted Date: 21 March 2025
Available online: 25 March 2025

Abstract

Human bocavirus 1 (HBoV1; family: Parvoviridae) causes a wide spectrum of respiratory diseases in children and gastroenteritis in adults. A lack of sensitive cell lines and efficient animal models hinders research on HBoV, including the development of anti-HBoV drugs or vaccines. Although the construction of a wild-type HBoV1 infectious clone has been reported, generating HBoV1 infectious clone carrying foreign reporter genes with suitable insertion sites in its genome while retaining replicative ability remains challenging. Here, HBoV1 infectious clones harboring the 11-amino-acid HiBiT tag at five distinct insertion sites were constructed and evaluated. Only the recombinant HBoV1 carrying the HiBiT tag in the N-terminus of the NS1 protein (HBoV1-HiBiT_NS1) displayed comparable characteristics to wild-type HBoV1 as determined via the analysis of viral DNA copy number, NanoLuc activity, viral protein expression, and the formation of replication intermediates. Notably, the replication kinetics of HBoV1-HiBiT_NS1 could be examined by monitoring NanoLuc activity, which was noted to be correlated with the viral DNA level. Additionally, we successfully applied HiBiT-tagged HBoV1 for the evaluation of antiviral drug activity and identified ivermectin (EC50 = 2.27 μM) as a potent anti-HBoV1 replication drug. Overall, our study demonstrated that the HBoV1-HiBiT_NS1 reporter can serve as a convenient platform for screening candidate drugs targeting HBoV1 replication and may also be useful for investigating the life cycle of the virus.
- Human bocavirus (HBoV)
- , HiBiT tag
- , Infectious clone
- , Antiviral drug evaluation

Electronic Supplementary Material

10.1016j.virs.2025.03.009-ESM.docx
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