title: DsRNA-mediated silencing of Nudix hydrolase in Trichinella spiralis inhibits the larval invasion and survival in mice.
abstract: The aim of this study was to investigate the functions of Trichinella spiralis Nudix hydrolase (TsNd) during the larval invasion of intestinal epithelial cells (IECs), development and survival in host by RNAi. The TsNd-specific double-stranded RNA (dsRNA) was designed to silence the expression of TsNd in T. spiralis larvae. DsRNA were delivered to the larvae by soaking incubation or electroporation. Silencing effect of TsNd transcription and expression was determined by real-time PCR and Western blotting, respectively. The infectivity of larvae treated with dsRNA was investigated by the in vitro larval invasion of IECs and experimental infection in mice. After being soaked with 40 ng/μl of dsRNA-TsNd, the transcription and expression level of TsNd gene was inhibited 65.8% and 56.4%, respectively. After being electroporated with 40 ng/μl of dsRNA-TsNd, the transcription and expression level of TsNd gene was inhibited 74.2% and 58.2%, respectively. Silencing TsNd expression by both soaking and electroporation inhibited significantly the larval invasion of IECs in a dose-dependent manner (r1 = -0.96798, r2 = -0.98707). Compared with the mice inoculated with untreated larvae, mice inoculated with larvae soaked with TsNd dsRNA displayed a 49.9% reduction in adult worms and 39.9% reduction in muscle larvae, while mice inoculated with larvae electroporated with TsNd dsRNA displayed a 83.4% reduction in adult worms and 69.5% reduction in muscle larvae, indicating that electroporation has a higher efficiency than soaking in inhibiting the larval development and survival in mice. Our results showed that silencing TsNd expression in T. spiralis inhibited significantly the larval invasion and survival in host.
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China.
authors.forename: Shuai Bing
authors.initials: SB
authors.lastname: Zhang
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Jiang
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China. Electronic address: wangzq@zzu.edu.cn.
authors.forename: Zhong Quan
authors.initials: ZQ
authors.lastname: Wang
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China.
authors.forename: Shao Rong
authors.initials: SR
authors.lastname: Long
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China.
authors.forename: Ruo Dan
authors.initials: RD
authors.lastname: Liu
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China.
authors.forename: Xi
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China.
authors.forename: Wei
authors.initials: W
authors.lastname: Yang
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China.
authors.forename: Hui Jun
authors.initials: HJ
authors.lastname: Ren
authors.affiliation: Department of Parasitology, Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou 450052, PR China. Electronic address: cuij@zzu.edu.cn.
authors.forename: Jing
authors.initials: J
authors.lastname: Cui
mesh_terms: D000818:Animals; D018274:Electroporation; D005260:Female; D005298:Fertility / Q000502:physiology; D007413:Intestinal Mucosa / Q000166:cytology / Q000469:parasitology; D007814:Larva / Q000201:enzymology / Q000235:genetics / Q000502:physiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011755:Pyrophosphatases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference / Q000502:physiology*; D012330:RNA, Double-Stranded / Q000096:biosynthesis / Q000302:isolation & purification / Q000502:physiology*; D018510:RNA, Helminth / Q000096:biosynthesis / Q000302:isolation & purification / Q000502:physiology; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D014158:Transcription, Genetic; D017160:Trichinella spiralis / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D000096964:Nudix Hydrolases
chemical_list: D011755:Pyrophosphatases; D012330:RNA, Double-Stranded; D018510:RNA, Helminth; D000096964:Nudix Hydrolases
keywords: Infectivity; Invasion; Nudix hydrolases; RNAi; Trichinella spiralis; dsRNA