ShanghaiTech University Knowledge Management System
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]) |
ISSN | 2379-5042 |
卷号 | 6期号:3 |
DOI | 10.1128/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 |
引用统计 | 正在获取...
|
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). |
条目包含的文件 | ||||||
文件名称/大小 | 文献类型 | 版本类型 | 开放类型 | 使用许可 |
修改评论
除非特别说明,本系统中所有内容都受版权保护,并保留所有权利。