Schistosoma japonicum: inhibition of Mago nashi gene expression by shRNA-mediated RNA interference.
RNA interference (RNAi) mediated by short interfering RNA (siRNA) is a powerful reverse genetics tool and holds enormous therapeutic potential for various diseases, including parasite infections. siRNAs bind their complementary mRNA and lead to degradation of their specific mRNA targets. RNAi has been widely used for functional analysis of specific genes in various cells and organisms. In this paper, we tested the potential of silencing the expression of the Mago nashi gene in Schistosoma japonicum by siRNAs derived from shRNA expressed by mammalian Pol III promoter H1. Schistosomula, transformed from cercariae by mechanical shearing of the tails, were electroporated with Mago nashi shRNA expression vector. Aliquots of parasites were harvested at days 1, 3, and 5 after electroporation, respectively. Levels of Mago nashi mRNA and protein were determined by RT-PCR and Western blotting analysis. The results showed that shRNA expressed from mammalian Pol III promoter H1 specifically reduced the levels of Mago nashi mRNA and proteins in S. japonicum. Changes in testicular lobes were apparent when parasites were introduced into mammalian hosts. Thus, vector-mediated gene silencing is applicable to S. japonicum, which provides a means for the functional analysis of genes in this organism.
Zhao
Zhi-rong
ZR
Department of Microbiology & Parasitology, Anhui Medical University, Meishan Road, Hefei 230032, China.
Lei
Li
L
Liu
Miao
M
Zhu
Shao-chun
SC
Ren
Cui-ping
CP
Wang
Xiao-nan
XN
Shen
Ji-jia
JJ
D000818:Animals; D015153:Blotting, Western; D018509:DNA, Helminth / Q000032:analysis; D018274:Electroporation; D005260:Female; D005786:Gene Expression Regulation / Q000235:genetics*; D015801:Helminth Proteins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D018510:RNA, Helminth / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012549:Schistosoma japonicum / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure
D018509:DNA, Helminth; D015801:Helminth Proteins; D009687:Nuclear Proteins; D018510:RNA, Helminth; D012333:RNA, Messenger