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	Characterization of cis-acting rna elements of zika virus by using a self-splicing ribozyme-dependent infectious clone
	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
	2017-11-01
卷号	91期号:21页码:14
关键词	Zika virus Self-splicing intron Infectious cdna clone Cis-acting rna elements Reverse genetics
ISSN号	0022-538X
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
内容类型	期刊论文
URI标识	http://www.corc.org.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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