title: Ascaris suum: RNAi mediated silencing of enolase gene expression in infective larvae.
abstract: Ascaris suum is an important parasite of pigs that causes tremendous economic losses globally to agriculture and animal husbandry annually. RNA interference (RNAi) technology has been described as a successful and useful approach for the elucidation of gene function in parasitic nematodes. In the present study, RNAi was used to silence the expression of a gene encoding enolase in A. suum by soaking infective larvae in double-stranded RNA derived from an EST (representing As-enol-1) selected from an A. suum infective larvae-specific cDNA library. The mRNA levels of RNAi-treated larvae were examined by Reverse-Transcription PCR (RT-PCR) analysis. The survival of RNAi-treated larvae was compared with larvae treated with dsRNA-free culture medium. The effect of enolase depletion on the development of A. suum larvae was assessed by infecting BALB/c mice with RNAi-treated larvae. The results showed that enolase gene expression was silenced completely and the survival rate of the RNAi-treated nematodes was reduced by 20.11% (P<0.01) after soaking for 72 h. Although no significant difference was detected in the numbers of larvae recovered from the liver and lungs of infected mice 4 days post infection, RNAi knockdown of the A. suum enolase mRNA led to significant shorter larvae, indicating that loss of enolase expression may cause delays in larval development.
authors.affiliation: College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, PR China.
authors.forename: Ning
authors.initials: N
authors.lastname: Chen
authors.forename: Min-Jun
authors.initials: MJ
authors.lastname: Xu
authors.forename: Alasdair J
authors.initials: AJ
authors.lastname: Nisbet
authors.forename: Cui-Qin
authors.initials: CQ
authors.lastname: Huang
authors.forename: Rui-Qing
authors.initials: RQ
authors.lastname: Lin
authors.forename: Zi-Guo
authors.initials: ZG
authors.lastname: Yuan
authors.forename: Hui-Qun
authors.initials: HQ
authors.lastname: Song
authors.forename: Xing-Quan
authors.initials: XQ
authors.lastname: Zhu
mesh_terms: D000818:Animals; D001196:Ascariasis / Q000469:parasitology; D017165:Ascaris suum / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D007814:Larva / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D008099:Liver / Q000469:parasitology; D008168:Lung / Q000469:parasitology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010641:Phenotype; D010751:Phosphopyruvate Hydratase / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013047:Specific Pathogen-Free Organisms; D013552:Swine
chemical_list: D012333:RNA, Messenger; D010751:Phosphopyruvate Hydratase