Establishment of an in vitro reporter system for screening HBx-targeting molecules

Establishment of an in vitro reporter system for

screening HBx-targeting molecules

Chenjian Gu;Shuai Tao;Kongying Hu;Lijun Ming;Mengjun Luo;Huimin Guo;Yu Su;Jing Liu;Youhua Xie

【期刊名称】《生物化学与生物物理学报：英文版》 【年(卷),期】2024(051)004

【摘要】Chronic hepatitis B virus (HBV) infection remains a global public health problem.HBV-encoded X protein (HBx) is a multifunctional regulator that is required to initiate and maintain productive HBV infection,and is involved in HBV-related hepatocellular carcinoma (HCC).Inhibitors that interfere with the functions of HBx could be useful not only for the inhibition of HBV replication but also for the prevention or treatment of HBV-related HCC.To screen molecules that target HBx on a large scale remains a technical challenge due to a lack of sensitive and high-throughput system.In this work,we established an in vitro bioluminescent

reporter

system

for

screening

HBx-targeting

molecules.The system is based on a secretory fusion protein that combines HBx and NanoLuc (HBx-Nluc).The measured activity of NanoLuc in the culture supernatant of HBx-Nluc-expressing cells directly reflects the level of secreted HBx-Nluc.HBx protein-targeting intracellular anti-HBx single-chain variable fragment and RNA-targeting shRNA significantly reduced the secreted NanoLuc activity in HBx-Nluc-

expressing cells.This system is simple and sensitive,and compatible with continuous non-disruptive screening,suggesting its potential usefulness for high-throughput screening and evaluating HBx-targeting molecules. 【总页数】10页(431-440) 【关键词】

【作者】Chenjian Gu;Shuai Tao;Kongying Hu;Lijun Ming;Mengjun Luo;Huimin Guo;Yu Su;Jing Liu;Youhua Xie

【作者单位】Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China;Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China;Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China;Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China;Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032,

China;Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China;Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China;Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China;Key Laboratory of Medical Molecular Virology(MOE/NHC), Department of Microbiology and Parasitology, School of Basic Medical Sciences, Shanghai Medical College, Fudan University, Shanghai 200032, China;Children's Hospital, Fudan University, Shanghai 201102, China 【正文语种】英文 【中图分类】 【文献来源】

https://www.zhangqiaokeyan.com/academic-journal-cn_acta-biochimica-biophysica-sinica_thesis/0201270256579.html 【相关文献】

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