Transient expression of DNA and RNA in parasitic helminths by using particle bombardment.
Parasitic helminths (worms belonging to several metazoan phyla) cause considerable morbidity and mortality in humans. They are an important veterinary problem, and they result in significant economic losses in animal grazing and agriculture. Experimental studies on parasitic helminths have been limited by a lack of parasite cell lines and methods for molecular genetic analyses. We evaluated particle bombardment (biolistics) as a strategy to introduce and express nucleic acids in these multicellular parasites. By using embryos of the parasitic nematode Ascaris as a model, we developed methods to introduce and express both DNA and RNA during several stages of Ascaris embryogenesis. Biolistic transfection will facilitate experimental strategies in Ascaris embryos complementing other biochemical tools available (e.g., in vitro whole-cell embryo extracts for transcription, RNA processing, and translation). Transfection experiments with adult schistosomes further suggest that the biolistic strategy should be applicable to a variety of other parasitic helminths. The development of these methods provides molecular genetic tools to study gene expression and the biology of a variety of types and developmental stages of important helminth parasites.
Department of Biological Sciences, Fordham University, Bronx, NY 10458, USA. rdavis@murray.fordham.edu
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Davis
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Parra
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LoVerde
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Ribeiro
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Glorioso
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Hodgson
D000818:Animals; D001200:Ascaris / Q000196:embryology* / Q000235:genetics; D019470:Biolistics / Q000379:methods*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005260:Female; D015870:Gene Expression; D017930:Genes, Reporter; D006046:Gold; D008156:Luciferases; D010316:Particle Size; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics*; D012547:Schistosoma / Q000235:genetics / Q000469:parasitology; D014158:Transcription, Genetic; D014162:Transfection
D012313:RNA; D006046:Gold; D004247:DNA; D008156:Luciferases