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RNAi-mediated silencing of enolase confirms its biological importance in Clonorchis sinensis

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Abstract

Clonorchis sinensis (C. sinensis) infection is still a common public health problem in freshwater fish consumption areas in Asian countries. More molecular evidence are required to speed up the prevention strategies to control this kind of infectious disease. In the present study, to confirm the biological importance of Csenolase followed by our previous observations of the key metabolic enzyme, we explored the RNA silence effect of the Csenolase-derived RNA interference (RNAi) in C. sinensis. The extramembranous region aa105–226 was selected as the target sequence of RNA silence. Csenolase-derived double strand RNA (dsRNA-Csenolase, 366 bp) was synthetized and delivered into C. sinensis by soaking approach. The penetration of dsRNA into adult worms and metacercariae was tracked using fluorescently labeled RNA. Western blotting and qRT-PCR experiments were performed to determine dsRNA-Csenolase-silencing effect. Our results showed that, after incubating for 120 h, dsRNA-Csenolase could effectively target and downregulate the expression of Csenolase in both adult worms (P < 0.001) and metacercariae (P < 0.01), resulting in a remarkable killing effect on C. sinensis adult worms (P < 0.01). Fluorescent Cy3-labeled dsRNA was mostly deposited in the uterus and vitellarium of adult worm and in the cyst wall of metacercaria. The present study is the first report of RNAi trials in C. sinensis, allowing further applications in identifying functional genes in C. sinensis.

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Acknowledgments

This work was supported by the National Key Basic Research and Development Project (973 project; no. 2010CB530000) and National Natural Science Foundation of China (no. 81171602) to XBY. The work was supported in part by the National Natural Science Foundation of China (no. 81101270) to YH.

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Authors and Affiliations

  1. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, 74th Zhongshan 2nd Road, Guangzhou, 510080, Guangdong Province, People’s Republic of China

    Xiaoyun Wang, Wenjun Chen, Yanli Tian, Yan Huang, Xuerong Li & Xinbing Yu

  2. Key Laboratory for Tropical Diseases Control of Ministry of Education, Sun Yat-sen University, 74th Zhongshan 2nd Road, Guangzhou, 510080, Guangdong Province, People’s Republic of China

    Xiaoyun Wang, Wenjun Chen, Yanli Tian, Yan Huang, Xuerong Li & Xinbing Yu

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  1. Xiaoyun Wang
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  2. Wenjun Chen
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  3. Yanli Tian
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  4. Yan Huang
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  5. Xuerong Li
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  6. Xinbing Yu
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Correspondence to Xinbing Yu.

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Wang, X., Chen, W., Tian, Y. et al. RNAi-mediated silencing of enolase confirms its biological importance in Clonorchis sinensis . Parasitol Res 113, 1451–1458 (2014). https://doi.org/10.1007/s00436-014-3785-0

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  • DOI: https://doi.org/10.1007/s00436-014-3785-0

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