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 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.
Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 518, Ziyue Road, Shanghai, 200241, People's Republic of China.
Yunxia
Y
Yang
Yamei
Y
Jin
Pingping
P
Liu
Yanli
Y
Shi
Yufan
Y
Cao
Jinming
J
Liu
Yaojun
Y
Shi
Hao
H
Li
Jiaojiao
J
Lin
D000818:Animals; D024062:Collagen Type V / Q000235:genetics / Q000378:metabolism*; D005260:Female; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D034622:RNA Interference; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction; D012549:Schistosoma japonicum / Q000033:anatomy & histology* / Q000235:genetics / Q000502:physiology*
D024062:Collagen Type V