title: In Vitro Gene Silencing of the Fish Microsporidian Heterosporis saurida by RNA Interference.
abstract: Heterosporis saurida, a microsporidian parasite of lizardfish, Saurida undosquamis, causes severe economic losses in marine aquaculture. Among the novel approaches being explored for treatment of parasitic infections in aquaculture is small interfering RNA molecules. The aim of the present study was to investigate the efficiency of using siRNA to knock down expression of specific genes of H. saurida in vitro. For this purpose, siRNAs specific for ATP/ADP antiporter 1 and methionine aminopeptidase II genes were designed and tested using a previously developed in vitro cultivation model. Silencing of H. saurida target genes was assessed and the efficacy of using siRNA for inhibition of gene expression was measured by quantitative real-time polymerase chain reaction (PCR). Silencing of ATP/ADP antiporter 1 or methionine aminopeptidase II by siRNA reduced H. saurida infection levels in EK-1 cells 40% and 60%, respectively, as measured by qRT-PCR and spore counts. Combined siRNA treatment of both ATP/ADP antiporter 1 and methionine aminopeptidase II siRNAs was more effective against H. saurida infection as seen by the 16S rRNA level and spore counts. Our study concluded that siRNA could be used to advance development of novel approaches to inhibit H. saurida and provide an alternative approach to combat microsporidia.
authors.affiliation: 1 Clinical Division of Fish Medicine, University of Veterinary Medicine , Vienna, Austria .
authors.forename: Mona
authors.initials: M
authors.lastname: Saleh
authors.affiliation: 1 Clinical Division of Fish Medicine, University of Veterinary Medicine , Vienna, Austria .
authors.forename: Gokhlesh
authors.initials: G
authors.lastname: Kumar
authors.affiliation: 2 Zoology Department, College of Science, King Saud University , Riyadh, Saudi Arabia .|3 Zoology Department, Faculty of Science, Beni-Suef University , Beni-Suef, Egypt .
authors.forename: Abdel-Azeem
authors.initials: AA
authors.lastname: Abdel-Baki
authors.affiliation: 2 Zoology Department, College of Science, King Saud University , Riyadh, Saudi Arabia .|4 Department of Zoology and Entomology, Faculty of Science, Helwan University , Cairo, Egypt .
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Dkhil
authors.affiliation: 1 Clinical Division of Fish Medicine, University of Veterinary Medicine , Vienna, Austria .
authors.forename: Mansour
authors.initials: M
authors.lastname: El-Matbouli
authors.affiliation: 2 Zoology Department, College of Science, King Saud University , Riyadh, Saudi Arabia .
authors.forename: Saleh
authors.initials: S
authors.lastname: Al-Quraishy
mesh_terms: D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000235:genetics; D000818:Animals; D017920:Antiporters / Q000037:antagonists & inhibitors / Q000235:genetics; D005393:Fish Diseases / Q000235:genetics / Q000382:microbiology / Q000517:prevention & control; D005399:Fishes / Q000235:genetics* / Q000382:microbiology; D020868:Gene Silencing; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000235:genetics; D016814:Microsporidia / Q000472:pathogenicity; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000494:pharmacology
chemical_list: D017920:Antiporters; D034741:RNA, Small Interfering; D000626:Aminopeptidases; C106541:methionine aminopeptidase 2; D008666:Metalloendopeptidases