Abstract
Type V collagen is a component of non-cartilaginous tissues and is important in the determination of fibril structure and matrix organization, although its functions are still poorly understood. In this report, RNA interference (RNAi) approaches were used to investigate the effects of knockdown of the schistosome type V collagen (SjColV) gene. In this study, three different short interfering (si) RNAs targeting different regions of the gene were designed to suppress the expression of SjColV in Schistosoma japonicum using a soaking method. By establishing controls for measuring off-target RNAi effects, we found that different siRNA sequences had different levels of effectiveness. Although all the siRNAs tested reduced SjColV transcript levels, the S1 siRNA consistently reduced SjColV expression to >99 % of the control. In the following experiments, S1 siRNA was adapted to inhibit SjColV expression, and the silencing effects were detected by real-time PCR and Western blot. The spawning and egg hatching of parasites were calculated, while the worms’ morphology was taken by scanning electron microscopy. The results show that silencing the expression of SjColV significantly affects the spawning and egg hatching of S. japonicum, and it also affects the worms’ morphology.
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Acknowledgments
We thank Zhiqiang Fu and Ke Lu of the Shanghai Veterinary Research Institute of Chinese Academy of Agricultural Sciences for their contribution to the parasite maintenance and cercariae isolation. This work was supported by the Chinese National Natural Science Foundation (No. 30800819 and 31072129), Shanghai Pujiang Program (No. 09PJ14118), and the Basic scientific research operation cost of state-leveled public welfare scientific research courtyard (No. 2012ZL091).
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Yang, Y., Jin, Y., Liu, P. et al. RNAi silencing of type V collagen in Schistosoma japonicum affects parasite morphology, spawning, and hatching. Parasitol Res 111, 1251–1257 (2012). https://doi.org/10.1007/s00436-012-2959-x
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DOI: https://doi.org/10.1007/s00436-012-2959-x