Transfection of Schistosoma mansoni by electroporation and the description of a new promoter sequence for transgene expression.
We sought to investigate the efficacy of electroporation for the introduction of plasmid-based DNA constructs into Schistosoma mansoni, and expanded our study to examine parameters governing transgene expression, including requirements of a 5' and 3' flanking sequence, as well as parasite developmental effects on transgene expression. We used luciferase as a reporter gene for this application. Our data show that electroporation allows the transfection of immature schistosomes, and defines 5' promoter sequence from the schistosome actin gene (SmAct1.1), coupled promiscuously with various 3' terminator sequences, as a powerful promoter of transgene expression in growing, but not early non-growing, schistosomula. The methodology described herein will facilitate ectopic expression of genes of interest in schistosomes.
Correnti
Jason M
JM
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, 318 Hill Pavilion, 380 South University Avenue, Philadelphia, PA 19104-4539, USA.
Jung
Euihye
E
Freitas
Tori C
TC
Pearce
Edward J
EJ
D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods*; D015870:Gene Expression; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012550:Schistosoma mansoni / Q000166:cytology / Q000235:genetics*; D013997:Time Factors; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics*