RNAi silencing of type V collagen in Schistosoma japonicum affects parasite morphology, spawning, and hatching

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.

Comparative Proteomics Reveals Characteristic Proteins on Praziquantel-resistance inSchistosoma mansoni

“…It has been suggested that ubiquitination is of interest in S. mansoni because this process could be a potential target for the design of new drugs (Guerra-Sá et al, 2005), being ubiquitin protein ligase E3a a good target to be studied. In regard to collagen alpha chain, Yang and colleagues (Yang et al, 2012)described that silencing the expression of a type of collagen (type V collagen) significantly affects the spawning and egg hatching of S. japonicum , and it also affects the morphology of the worms (Yang et al, 2012). Therefore, it would be very interesting evaluate the role of each of these proteins in PZQ-resistance, specially, collagen alpha chain, since it seems to occur morphological alterations in eggs and worms of S. mansoni PZQ-resistant strain (Pinto-Almeida et al, 2016).…”

Gaining biological perspectives from schistosome genomes

“…An RNAi study of collagen V was performed initially in in vitro maintained S. japonicum schistosomula (SjColV), subjected to a range of si-RNA strategies, so as to identify the most efficient knockdown [54]. The optimal si-RNA region was then used for in vivo knockdown of parasites.…”

New Frontiers inSchistosomaGenomics and Transcriptomics

“…Many proteases (cathepsins and aminopeptidases) [77–79], kinases [80], multidrug resistance transporters [81], and type V collagen [82], among other proteins have been studied in diverse life stages of S. japonicum and S. mansoni .…”