title: Inhibition of Japanese encephalitis virus NS1 protein expression in cell by small interfering RNAs.
abstract: Japanese encephalitis virus (JEV), a serious mosquitoborne flavivirus, causes an acute infection of the central system resulting in encephalitis of humans and many kinds of animals. A high proportion of the survivors exhibit neurogical and psychiatric sequelae. NS1 is one of important non-structural proteins, which was found to be associated with viral RNA replication. To inhibit NS1 expression, four small interfering RNAs (siRNAs) expression plasmids (pS-NS1A, pS-NS1B, pS-NS1C and pS-NS1D) were generated to target four different coding regions of the NS1 gene, and were separately co-transfected into Vero cells with an NS1-EGFP fusion expression plasmid pNS1-EGFP. NS1 expression was evaluated by fluorescence microscope, flow cytometry assay, Western blot and RT-PCR. The results revealed that pS-NS1B, pS-NS1C and pS-NS1D could effectively and specifically inhibit NS1 expression in Vero cells. Our data suggested that these siRNAs could be used to inhibit JEV replication by silencing NS1 protein expression in further study.
authors.affiliation: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, Hubei, 430070, P.R China.
authors.forename: Xueqin
authors.initials: X
authors.lastname: Liu
authors.forename: Shengbo
authors.initials: S
authors.lastname: Cao
authors.forename: Rui
authors.initials: R
authors.lastname: Zhou
authors.forename: Gaoyuan
authors.initials: G
authors.lastname: Xu
authors.forename: Shaobo
authors.initials: S
authors.lastname: Xiao
authors.forename: Ying
authors.initials: Y
authors.lastname: Yang
authors.forename: Minxuan
authors.initials: M
authors.lastname: Sun
authors.forename: Yaoming
authors.initials: Y
authors.lastname: Li
authors.forename: Huanchun
authors.initials: H
authors.lastname: Chen
mesh_terms: D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D004664:Encephalitis Virus, Japanese / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D034741:RNA, Small Interfering / Q000096:biosynthesis / Q000502:physiology*; D013698:Templates, Genetic; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors* / Q000096:biosynthesis* / Q000235:genetics
chemical_list: C100315:NS1 protein, Flavivirus; D034741:RNA, Small Interfering; D017361:Viral Nonstructural Proteins