Polyethyleneimine Mediated DNA Transfection in Schistosome Parasites and Regulation of the WNT Signaling Pathway by a Dominant-Negative SmMef2

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Abstract
Schistosomiasis is a serious global problem and the second most devastating parasitic disease following malaria. Parasitic worms of the genus Schistosoma are the causative agents of schistosomiasis and infect more than 240 million people worldwide. The paucity of molecular tools to manipulate schistosome gene expression has made an understanding of genetic pathways in these parasites difficult, increasing the challenge of identifying new potential drugs for treatment. Here, we describe the use of a formulation of polyethyleneimine (PEI) as an alternative to electroporation for the efficacious transfection of genetic material into schistosome parasites. We show efficient expression of genes from a heterologous CMV promoter and from the schistosome Sm23 promoter. Using the schistosome myocyte enhancer factor 2 (SmMef2), a transcriptional activator critical for myogenesis and other developmental pathways, we describe the development of a dominant-negative form of the schistosome Mef2. Using this mutant, we provide evidence that SmMef2 may regulate genes in the WNT pathway. We also show that SmMef2 regulates its own expression levels. These data demonstrate the use of PEI to facilitate effective transfection of nucleic acids into schistosomes, aiding in the study of schistosome gene expression and regulation, and development of genetic tools for the characterization of molecular pathways in these parasites. Schistosomiasis is a global disease infecting more than 240 million people worldwide and is ranked second only to malaria in global health importance. The causative agents of human schistosomiasis are parasitic worms that ingest red blood cells and can live for decades producing hundreds of eggs daily. There is one primary drug for treatment of schistosomiasis, but its use for over 30 years has raised concern over the development of drug resistance and thus created a need for new drugs. A challenge to the rational development of effective antischistosomals has been the difficulty in manipulating schistosome gene expression, and thus a limitation in our understanding of schistosome gene function. Here, we present a new and straightforward method for inserting genes into schistosomes and expressing them. In addition, to our knowledge we provide the first example of dominant negative gene expression to modify transcriptional regulation using a molecular genetics approach to study this globally important parasite.

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