title: The siRNA-mediated silencing of Trichinella spiralis nudix hydrolase results in reduction of larval infectivity.
abstract: Previous studies showed that Trichinella spiralis Nudix hydrolase (TsNd) bound to intestinal epithelial cells (IECs), and vaccination of mice with rTsNd or TsNd DNA produced a partial protective immunity against T. spiralis infection. In this study, three TsNd specific small interfering RNA (siRNA) were designed to silence the expression of TsNd in T. spiralis larvae. SiRNAs were delivered to the larvae by electroporation. Silencing effect of TsNd transcription and expression was determined by real-time PCR and Western blotting, respectively. The infectivity of the larvae treated with siRNA was investigated by the in vitro larval invasion of IECs and experimental infection in mice. The results showed that siRNAs were efficiently delivered into T. spiralis larvae through electroporation. Real-time PCR and Western blotting showed that transcription and expression level of TsNd gene was inhibited 73.3 and 76.7 %, respectively, after being electroporated with 2 μM of siRNA-275 for 1 day. Silencing TsNd expression inhibited significantly the larval invasion of IECs (P < 0.01) and was in a dose-dependent manner (r = -0.97941). The mice with infected larvae treated with TsNd siRNA displayed a 63.6 % reduction in intestinal adult worms and 68.8 % reduction in muscle larval burden compared with mice infected with control siRNA-treated larvae. Our results showed that silencing TsNd expression in T. spiralis significantly reduced the larval infectivity and survival in host.
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou, 450052, China, wangzq@zzu.edu.cn.
authors.forename: Zhong Quan
authors.initials: ZQ
authors.lastname: Wang
authors.forename: Shuai Bing
authors.initials: SB
authors.lastname: Zhang
authors.forename: Peng
authors.initials: P
authors.lastname: Jiang
authors.forename: Ruo Dan
authors.initials: RD
authors.lastname: Liu
authors.forename: Shao Rong
authors.initials: SR
authors.lastname: Long
authors.forename: Xi
authors.initials: X
authors.lastname: Zhang
authors.forename: Hui Jun
authors.initials: HJ
authors.lastname: Ren
authors.forename: Jing
authors.initials: J
authors.lastname: Cui
mesh_terms: D000818:Animals; D004847:Epithelial Cells / Q000276:immunology / Q000378:metabolism; D005260:Female; D007413:Intestinal Mucosa / Q000378:metabolism; D007422:Intestines / Q000276:immunology; D007814:Larva; D051379:Mice; D008807:Mice, Inbred BALB C; D011755:Pyrophosphatases / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D013047:Specific Pathogen-Free Organisms; D017160:Trichinella spiralis / Q000201:enzymology* / Q000235:genetics / Q000276:immunology / Q000502:physiology; D014235:Trichinellosis / Q000276:immunology / Q000469:parasitology / Q000517:prevention & control*; D014611:Vaccination; D000096964:Nudix Hydrolases
chemical_list: D011755:Pyrophosphatases; D034741:RNA, Small Interfering; D000096964:Nudix Hydrolases