An improved and secreted luciferase reporter for schistosomes.
Schistosomes are multicellular parasites of humans exhibiting interesting biological adaptations to their parasitic lifestyle. Concerted and in depth analyses of these adaptations and their cell and molecular biology requires further development of molecular genetic tools in schistosomes. In the current study, we demonstrate that a Gaussia luciferase reporter leads to significantly higher levels of luciferase activity in schistosomes compared to other tested luciferases. In addition, Gaussia luciferase can be secreted into culture media enabling non-invasive analysis of reporter activity. The secretion of Gaussia luciferase should allow a variety of new experimental paradigms for schistosome studies. Comparison of biolistic and electroporation transfection methods using luciferase RNA reporters and the luciferase acitivty produced indicates that electroporation of sporocysts and schistosomula is the most efficient transfection method for the four stages analyzed. These data should facilitate additional studies in schistosomes and provide a framework for further development of DNA transfection and gene expression analysis.
Cheng
Guofeng
G
Department of Pediatrics and Biochemistry, University of Colorado School of Medicine, Aurora, CO 80045, USA.
Davis
Richard E
RE
D000818:Animals; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D010274:Parasitology / Q000379:methods*; D012547:Schistosoma / Q000235:genetics*; D014162:Transfection / Q000379:methods
D008156:Luciferases