Integration of HiBiT into enteroviruses:A universal tool for advancing enterovirus virology research  

作　　者：Rui Yu Xiaohong Li Peng Zhang Minghao Xu Jitong Zhao Jingjing Yan Chenli Qiu Jiayi Shu Shuo Zhang Miaomiao Kang Xiaoyan Zhang Jianqing Xu Shuye Zhang 

机构地区：[1]School of Medicine,Shanghai University,Shanghai,200444,China [2]Clinical Center for Biotherapy,Zhongshan Hospital,Fudan University,Shanghai,200433,China [3]Guangzhou Institutes of Biomedicine and Health,The Chinese Academy of Sciences,Guangzhou,510530,China [4]Institutes of Biomedical Sciences,Fudan University,Shanghai,200032,China [5]Shanghai Geriatric Medical Center,Shanghai,201104,China [6]Clinical Center for Biotherapy,Zhongshan Hospital/Zhongshan Hospital(Xiamen),Fudan University,361015,China [7]Shanghai Public Health Clinical Center,Fudan University,Shanghai,201508,China

出　　处：《Virologica Sinica》2024年第3期422-433,共12页中国病毒学（英文版）

基　　金：supported by the National Natural Science Foundation of China(grant nos.82002135 and 82172250).

摘　　要：The utilization of enteroviruses engineered with reporter genes serves as a valuable tool for advancing our understanding of enterovirus biology and its applications,enabling the development of effective therapeutic and preventive strategies.In this study,our initial attempts to introduce a NanoLuc luciferase(NLuc)reporter gene into recombinant enteroviruses were unsuccessful in rescuing viable progenies.We hypothesized that the size of the inserted tag might be a determining factor in the rescue of the virus.Therefore,we inserted the 11-amino-acid HiBiT tag into the genomes of enterovirus A71(EV-A71),coxsackievirus A10(CVA10),coxsackievirus A7(CVA7),coxsackievirus A16(CVA16),namely EV-A71-HiBiT,CVA16-HiBiT,CVA10-HiBiT,CVA7-HiBiT,and observed that the HiBiT-tagged viruses exhibited remarkably high rescue efficiency.Notably,the HiBiT-tagged enteroviruses displayed comparable characteristics to the wild-type viruses.A direct comparison between CVA16-NLuc and CVA16-HiBiT recombinant viruses revealed that the tiny HiBiT insertion had minimal impact on virus infectivity and replication kinetics.Moreover,these HiBiT-tagged enteroviruses demonstrated high genetic stability in different cell lines over multiple passages.In addition,the HiBiT-tagged viruses were successfully tested in antiviral drug assays,and the sensitivity of the viruses to drugs was not affected by the HiBiT tag.Ultimately,our findings provide definitive evidence that the integration of HiBiT into enteroviruses presents a universal,convenient,and invaluable method for advancing research in the realm of enterovirus virology.Furthermore,HiBiT-tagged enteroviruses exhibit great potential for diverse applications,including the development of antivirals and the elucidation of viral infection mechanisms.

关 键 词：ENTEROVIRUS Infectious clones Reporter genes LUCIFERASE Drug screening 

分 类 号：R373.2[医药卫生—病原生物学]