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A stable and efficient infectious clone of ZIKV is urgently needed. However, the instability and toxicity of flavivirus cDNA clones in\n                    <jats:named-content content-type=\"genus-species\">Escherichia coli<\/jats:named-content>\n                    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\n                    <jats:italic>P.li.LSUI2<\/jats:italic>\n                    near the junction between the E and NS1 genes, which were removed from the RNA transcripts by an easy-to-establish\n                    <jats:italic>in vitro<\/jats:italic>\n                    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\n                    <jats:italic>cis<\/jats:italic>\n                    -acting RNA elements in ZIKV genomic RNA were identified based on this novel reverse genetics system, and the critical role of 5\u2032-SLA promoter and 5\u2032-3\u2032 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.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>IMPORTANCE<\/jats:bold>\n                    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\n                    <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n                    host. Moreover, two crucial\n                    <jats:italic>cis<\/jats:italic>\n                    -acting replication elements (5\u2032-SLA and 5\u2032-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.\n                  <\/jats:p>","DOI":"10.1128\/jvi.00484-17","type":"journal-article","created":{"date-parts":[[2017,8,16]],"date-time":"2017-08-16T20:30:18Z","timestamp":1502915418000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":43,"title":["Characterization of\n                    <i>cis<\/i>\n                    -Acting RNA Elements of Zika Virus by Using a Self-Splicing Ribozyme-Dependent Infectious Clone"],"prefix":"10.1128","volume":"91","author":[{"given":"Zhong-Yu","family":"Liu","sequence":"first","affiliation":[{"name":"State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, 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