Abstract
An in vivo, non-invasive technique for gene silencing by RNA interference (RNAi) in the snail, Biomphalaria glabrata, has been developed using cationic polymer polyethyleneimine (PEI) mediated delivery of long double-stranded (ds) and small interfering (si) RNA. Cellular delivery was evaluated and optimized by using a ‘mock’ fluorescent siRNA. Subsequently, we used the method to suppress expression of Cathepsin B (CathB) with either the corresponding siRNA or dsRNA of this transcript. In addition, the knockdown of peroxiredoxin (Prx) at both RNA and protein levels was achieved with the PEI-mediated soaking method. B. glabrata is an important snail host for the transmission of the parasitic digenean platyhelminth, Schistosoma mansoni that causes schistosomiasis in the neotropics. Progress is being made to realize the genome sequence of the snail and to uncover gene expression profiles and cellular pathways that enable the snail to either prevent or sustain an infection. Using PEI complexes, a convenient soaking method has been developed, enabling functional gene knockdown studies with either dsRNA or siRNA. The protocol developed offers a first whole organism method for host-parasite gene function studies needed to identify key mechanisms required for parasite development in the snail host, which ultimately are needed as points for disrupting this parasite mediated disease.
Highlights
Biomphalaria glabrata is an intermediate snail host that transmits the digenean platyhelminth parasite, Schistosoma mansoni, in the Western Hemisphere
Genesilencing technology involving the use of dsRNA have used an injection route to disrupt gene translation in the Schistosoma mansoni snail host, Biomphalaria glabrata in an effort to investigate how inhibition of various transcripts can affect the dynamics of the snail/parasite interaction
To make the use of gene silencing technology more widely applicable to functional gene studies in snails, we have developed a more convenient soaking method that uses a cationic carrier polyethylene amine (PEI) to deliver dsRNA or siRNA into juvenile snails
Summary
Biomphalaria glabrata is an intermediate snail host that transmits the digenean platyhelminth parasite, Schistosoma mansoni, in the Western Hemisphere. It is hoped that all these advances will lead eventually to the development of novel tools for halting infection at the snail stage of the parasite’s life cycle. For this disease transmission blocking strategy to come to fruition, we need a better understanding of what genes/cellular pathways in the snail host can be interfered with to bring about subsequent disruption of the parasite’s development
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