Development of a New Reverse Genetics System for Ebola Virus
2021-05
发表期刊MSPHERE (IF:3.7[JCR-2023],4.0[5-Year])
ISSN2379-5042
卷号6期号:3
DOI10.1128/mSphere.00235-21
摘要Ebola virus (EBOV) is a highly pathogenic negative-stranded RNA virus that has caused several deadly endemics in the past decades. EBOV reverse genetics systems are available for studying live viruses under biosafety level 4 (BSL-4) or subviral particles under BSL-2 conditions. However, these systems all require cotransfection of multiple plasmids expressing viral genome and viral proteins essential for EBOV replication, which is technically challenging and unable to naturally mimic virus propagation using the subviral particle. Here, we established a new EBOV reverse genetics system only requiring transfection of a single viral RNA genome into an engineered cell line that stably expresses viral nucleoprotein (NP), viral protein 35 (VP35), VP30, and large (L) proteins and has been fine-tuned for its superior permissiveness for EBOV replication. Using this system, subviral particles expressing viral VP40, glycoprotein (GP), and VP24 could be produced and continuously propagated and eventually infect the entire cell population. We demonstrated the authentic response of the subviral system to antivirals and uncovered that the VP35 amount is critical for optimal virus replication. Furthermore, we showed that fully infectious virions can be efficiently rescued by delivering the full-length EBOV genome into the same supporting cell, and the efficiency is not affected by genome polarity or virus variant specificity. In summary, our work provides a new tool for studying EBOV under different biosafety levels. IMPORTANCE Ebola virus is among the most dangerous viral pathogens, with a case fatality rate of up to 90%. Since 2013, the two largest and most complex Ebola outbreaks in Africa have revealed the lack of investigation on this notorious virus. A reverse genetics system is an important tool for studying viruses by producing mutant viruses or generating safer and convenient model systems. Here, we developed an EBOV life cycle modeling system in which subviral particles can spontaneously propagate in cell culture. In addition, this system can be employed to rescue infectious virions of homologous or heterologous EBOV isolates using either sense or antisense viral RNA genomes. In summary, we developed a new tool for EBOV research.
关键词Ebola virus filovirus negative-stranded RNA virus reverse genetics
收录类别SCIE
语种英语
WOS研究方向Microbiology
WOS类目Microbiology
WOS记录号WOS:000663083400010
出版者AMER SOC MICROBIOLOGY
原始文献类型Article
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文献类型期刊论文
条目标识符https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/127580
专题生命科学与技术学院_特聘教授组_钟劲组
生命科学与技术学院_硕士生
通讯作者Yan, Huimin; Zhong, Jin
作者单位
1.Chinese Acad Sci, Inst Pasteur Shanghai, Ctr Biosafety Mega Sci, Unit Viral Hepatitis,CAS Key Lab Mol Virol & Immu, Shanghai, Peoples R China;
2.Guangzhou Women & Childrens Med Ctr, Joint Lab Translat Precis Med, Guangzhou, Peoples R China;
3.Chinese Acad Sci, Wuhan Inst Virol, Wuhan, Peoples R China;
4.Chinese Acad Sci, Ctr Biosafety Mega Sci, Wuhan Natl Biosafety Lab, Wuhan, Peoples R China;
5.Chinese Acad Sci, Wuhan Inst Virol, Mucosal Immun Res Grp, State Key Lab Virol, Wuhan, Peoples R China;
6.Univ Chinese Acad Sci, Beijing, Peoples R China;
7.ShanghaiTech Univ, Shanghai, Peoples R China
通讯作者单位上海科技大学
推荐引用方式
GB/T 7714
Gan, Tianyu,Zhou, Dihan,Huang, Yi,et al. Development of a New Reverse Genetics System for Ebola Virus[J]. MSPHERE,2021,6(3).
APA Gan, Tianyu.,Zhou, Dihan.,Huang, Yi.,Xiao, Shuqi.,Ma, Ziyue.,...&Zhong, Jin.(2021).Development of a New Reverse Genetics System for Ebola Virus.MSPHERE,6(3).
MLA Gan, Tianyu,et al."Development of a New Reverse Genetics System for Ebola Virus".MSPHERE 6.3(2021).
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