Ascaris suum: RNAi mediated silencing of enolase gene expression in infective larvae

作者:Chen, Ning; Xu, Min-Jun; Ni**et, Alasdair J.; Huang, Cui-Qin; Lin, Rui-Qing; Yuan, Zi-Guo; Song, Hui-Qun; Zhu, Xing-Quan*
来源:Experimental Parasitology, 2011, 127(1): 142-146.
DOI:10.1016/j.exppara.2010.07.019

摘要

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.