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
Hauptverfasser: Yang, Yunxia, Jin, Yamei, Liu, Pingping, Shi, Yanli, Cao, Yufan, Liu, Jinming, Shi, Yaojun, Li, Hao, Lin, Jiaojiao
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container_issue 3
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container_title Parasitology research (1987)
container_volume 111
creator Yang, Yunxia
Jin, Yamei
Liu, Pingping
Shi, Yanli
Cao, Yufan
Liu, Jinming
Shi, Yaojun
Li, Hao
Lin, Jiaojiao
description 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.
doi_str_mv 10.1007/s00436-012-2959-x
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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 &gt;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. 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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 &gt;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.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer-Verlag</pub><pmid>22638918</pmid><doi>10.1007/s00436-012-2959-x</doi><tpages>7</tpages></addata></record>
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subjects Animals
Biological and medical sciences
Biomedical and Life Sciences
Biomedicine
Collagen
Collagen Type V - genetics
Collagen Type V - metabolism
Female
Fundamental and applied biological sciences. Psychology
General aspects
General aspects and techniques. Study of several systematic groups. Models
Immunology
Invertebrates
Male
Medical Microbiology
Mice
Mice, Inbred BALB C
Microbiology
Original Paper
Rabbits
Real-Time Polymerase Chain Reaction
RNA
RNA Interference
Schistosoma japonicum
Schistosoma japonicum - anatomy & histology
Schistosoma japonicum - genetics
Schistosoma japonicum - physiology
title RNAi silencing of type V collagen in Schistosoma japonicum affects parasite morphology, spawning, and hatching
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