title: RNA interference induces effective inhibition of mRNA accumulation and protein expression of SHEV ORF3 gene in vitro.
abstract: RNA interference (RNAi) provides a powerful promising approach to inhibit viral infection specifically. To explore the possibility of using RNAi as a strategy against HEV infection, which is a serious public health problem in developing countries of Asia, Middle East, Africa, and in Mexico, after the fusion protein expression plasmids pEGFP-ORF3 which contain the EGFP reporter gene and SHEV ORF3 as silencing target, were constructed; EGFP-ORF3 fusion protein expressing HEK 293 cell lines were established; and four siRNAs targeting ORF3 gene were designed, synthesized, and used to transfect the stable cell lines. At 24, 48, and 72 h after transfection, flow cytometry, real-time quantitative PCR, and Western blot were used to assess the expression level of ORF3. The results demonstrated that specific siRNAs which are sequence dependant effectively inhibited mRNA accumulation and protein expression of SHEV ORF3 in HEK 293 cells. These findings provide useful information for the development of RNAi-based prophylaxis and therapy for SHEV infection.
authors.affiliation: Hainan Key Lab of Tropical Animal Reproduction & Breeding and Epidemic Disease Research (Construction Period), Animal Genetic Engineering Key Lab of Haikou, College of Agriculture, Hainan University, Haidian Island, Haikou 570228, People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.forename: Ming
authors.initials: M
authors.lastname: Lei
authors.forename: Hanwei
authors.initials: H
authors.lastname: Jiao
authors.forename: Li
authors.initials: L
authors.lastname: Du
authors.forename: Ying
authors.initials: Y
authors.lastname: Cheng
authors.forename: Donglin
authors.initials: D
authors.lastname: Zhang
authors.forename: Yongchang
authors.initials: Y
authors.lastname: Hao
authors.forename: Churiga
authors.initials: C
authors.lastname: Man
authors.forename: Fengyang
authors.initials: F
authors.lastname: Wang
mesh_terms: D000818:Animals; D004197:Disease Reservoirs / Q000821:virology*; D015536:Down-Regulation; D015967:Gene Expression Regulation, Viral; D057809:HEK293 Cells; D016751:Hepatitis E / Q000821:virology*; D016752:Hepatitis E virus / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D016366:Open Reading Frames; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics / Q000378:metabolism; D013552:Swine / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012367:RNA, Viral; D014764:Viral Proteins