Ascaris suum: RNAi mediated silencing of enolase gene expression in infective larvae.
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.
Chen
Ning
N
College of Veterinary Medicine, South China Agricultural University, Guangzhou, Guangdong Province 510642, PR China.
Xu
Min-Jun
MJ
Nisbet
Alasdair J
AJ
Huang
Cui-Qin
CQ
Lin
Rui-Qing
RQ
Yuan
Zi-Guo
ZG
Song
Hui-Qun
HQ
Zhu
Xing-Quan
XQ
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
D012333:RNA, Messenger; D010751:Phosphopyruvate Hydratase