RNAi silencing of type V collagen in Schistosoma japonicum affects parasite morphology, spawning, and hatching
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 t...
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Veröffentlicht in: | Parasitology research (1987) 2012-09, Vol.111 (3), p.1251-1257 |
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Format: | Artikel |
Sprache: | eng |
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Zusammenfassung: | 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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ISSN: | 0932-0113 1432-1955 |
DOI: | 10.1007/s00436-012-2959-x |