Characterization of cis-acting rna elements of zika virus by using a self-splicing ribozyme-dependent infectious clone
文献类型：期刊论文
| 作者 | Liu, Zhong-Yu1; Yu, Jiu-Yang1; Huang, Xing-Yao1; Fan, Hang1; Li, Xiao-Feng1; Deng, Yong-Qiang1; Ji, Xue1; Cheng, Meng-Li1,2; Ye, Qing1; Zhao, Hui1 |
| 刊名 | Journal of virology
 |
| 出版日期 | 2017-11-01 |
| 卷号 | 91期号:21页码:14 |
| ISSN号 | 0022-538X |
| 关键词 | Zika virus Self-splicing intron Infectious cdna clone Cis-acting rna elements Reverse genetics |
| DOI | 10.1128/jvi.00484-17 |
| 通讯作者 | Qin, cheng-feng(qincf@bmi.ac.cn) |
| 英文摘要 | Zika virus (zikv) has caused significant outbreaks and epidemics in the americas recently, raising global concern due to its ability to cause microcephaly and other neurological complications. a stable and efficient infectious clone of zikv is urgently needed. however, the instability and toxicity of flavivirus cdna clones in escherichia coli hosts has hindered the development of zikv infectious clones. here, using a novel self-splicing ribozyme-based strategy, we generated a stable infectious cdna clone of a contemporary zikv strain imported from venezuela to china in 2016. the constructed clone contained a modified version of the group ii self-splicing intron p.li.lsui2 near the junction between the e and ns1 genes, which were removed from the rna transcripts by an easy-to-establish in vitro splicing reaction. transfection of the spliced rnas into bhk-21 cells led to the production of infectious progeny virus that resembled the parental virus. finally, potential cis-acting rna elements in zikv genomic rna were identified based on this novel reverse genetics system, and the critical role of 5'-sla promoter and 5'-3' cyclization sequences were characterized by a combination of different assays. our results provide another stable and reliable reverse genetics system for zikv that will help study zikv infection and pathogenesis, and the novel self-splicing intron-based strategy could be further expanded for the construction of infectious clones from other emerging and reemerging flaviviruses. importance the ongoing zika virus (zikv) outbreaks have drawn global concern due to the unexpected causal link to fetus microcephaly and other severe neurological complications. the infectious cdna clones of zikv are critical for the research community to study the virus, understand the disease, and inform vaccine design and antiviral screening. a panel of existing technologies have been utilized to develop zikv infectious clones. here, we successfully generated a stable infectious clone of a 2016 zikv strain using a novel self-splicing ribozyme-based technology that abolished the potential toxicity of zikv cdna clones to the e. coli host. moreover, two crucial cis-acting replication elements (5'-sla and 5'-cs) of zikv were first identified using this novel reverse genetics system. this novel self-splicing ribozyme-based reverse genetics platform will be widely utilized in future zikv studies and provide insight for the development of infectious clones of other emerging viruses. |
| WOS关键词 | SUBGENOMIC FLAVIVIRUS RNA ; ALGA PYLAIELLA-LITTORALIS ; DENGUE-VIRUS ; CODING REGION ; SECONDARY STRUCTURE ; GENOME CYCLIZATION ; CDNA-CLONE ; REPLICATION ; VACCINE ; INTRON |
| WOS研究方向 | Virology |
| WOS类目 | Virology |
| 语种 | 英语 |
| 出版者 | AMER SOC MICROBIOLOGY |
| WOS记录号 | WOS:000413195400003 |
| URI标识 | http://www.irgrid.ac.cn/handle/1471x/2373398 |
| 专题 | 武汉病毒研究所 |
| 通讯作者 | Qin, Cheng-Feng |
| 作者单位 | 1.Beijing Inst Microbiol & Epidemiol, State Key Lab Pathogen & Biosecur, Beijing, Peoples R China 2.Anhui Med Univ, Hefei, Anhui, Peoples R China 3.Chinese Acad Sci, Wuhan Inst Virol, Ctr Emerging Infect Dis, CAS Key Lab Special Pathogens & Biosafety, Wuhan, Hubei, Peoples R China |
| 推荐引用方式 GB/T 7714 | Liu, Zhong-Yu,Yu, Jiu-Yang,Huang, Xing-Yao,et al. Characterization of cis-acting rna elements of zika virus by using a self-splicing ribozyme-dependent infectious clone[J]. Journal of virology,2017,91(21):14. |
| APA | Liu, Zhong-Yu.,Yu, Jiu-Yang.,Huang, Xing-Yao.,Fan, Hang.,Li, Xiao-Feng.,...&Qin, Cheng-Feng.(2017).Characterization of cis-acting rna elements of zika virus by using a self-splicing ribozyme-dependent infectious clone.Journal of virology,91(21),14. |
| MLA | Liu, Zhong-Yu,et al."Characterization of cis-acting rna elements of zika virus by using a self-splicing ribozyme-dependent infectious clone".Journal of virology 91.21(2017):14. |
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来源：武汉病毒研究所
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