title: Inhibition of porcine reproductive and respiratory syndrome virus by specific siRNA targeting Nsp9 gene.
abstract: To screen siRNAs for effectively inhibiting the replication of porcine reproductive and respiratory syndrome virus (PRRSV). Four pairs of siRNA targeting Nsp9 gene of PRRSV and one non-efficient pair used as control were designed, synthesized and cloned into pSilencer4.1-CMV neo, designated as pSi-294, pSi-367, pSi-409, pSi-1488, pSi-Ctr. The recombinant plasmids were transfected into Marc-145 cells and infected with PRRSV 24h post transfection. Subsequently, IFA, real-time PCR, TCID50 and western blot were used for evaluating the inhibitory effect of the siRNA. IFA and western-blot results showed that pSi-294, pSi-1488 can effectively inhibit the expression of Nsp9 and M protein of PRRSV, real-time PCR result showed that the expression of Nsp9 gene were decreased from 86.56% to 93.66% compared to the negative control. siRNAs can be used as candidates for basic research of PRRSV.
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Jiexiong
authors.initials: J
authors.lastname: Xie
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Han
authors.initials: H
authors.lastname: Zhou
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Cui
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Chen
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Minze
authors.initials: M
authors.lastname: Zhang
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Shengchao
authors.initials: S
authors.lastname: Deng
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Pei
authors.initials: P
authors.lastname: Zhou
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Su
authors.affiliation: MOA Key Laboratory of Animal Vaccine Development, Ministry of China, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China. Electronic address: guihongzh@scau.edu.cn.
authors.forename: Guihong
authors.initials: G
authors.lastname: Zhang
mesh_terms: D003588:Cytopathogenic Effect, Viral; D015967:Gene Expression Regulation, Viral; D005814:Genes, Viral; D008958:Models, Molecular; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics*; D011487:Protein Conformation; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics; D014779:Virus Replication / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D017361:Viral Nonstructural Proteins
keywords: Nsp9; PRRSV; RNAi