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Biolistic Gene Transfer to Generate Transgenic Schistosomes, Rev A

onstrated that transgene expression can be monitored easily in living schistosomes. GFP activity was localized in the tegumental area of adult worms after heat shock. Interestingly, fluorescence was detected inside the larvae, indicating a different mechanism of HSP70 regulation in this life stage.

With the particle bombardment protocol presented here, we show a practical way to study genes in a homologous system. This method makes it possible to characterize regulatory elements of genes (Finken and Kunz 1997), to elucidate tissue-specific gene expression (Quijada et al. 2000), and to investigate the consequences of overexpression (Gillingham and Munro 2000). Since gold particles can be coated with RNA (Davis et al. 1999), it is feasible to design experiments for the silencing of genes using RNA interference (Bass 2000, Tavernarakis et al. 2000). Furthermore, it should be possible to use this transformation method not only for schistosomes but also for other multicellular parasites.


References
Bass BL, Double-stranded RNA as a template for gene silencing, Cell 101, 235238 (2000)

Buono RJ and Linser PJ, Transient expression of RSVCAT in transgenic zebrafish made by electroporation, Mol Mar Biol Biotechnol 1, 271275 (1992)

Davis RE et al., Transient expression of DNA and RNA in parasitic helminths by using particle bombardment, Proc Natl Acad Sci USA 96, 86878692 (1999)

Finken-Eigen M and Kunz W, Schistosoma mansoni: gene structure and locatlization of a homologue to cysteine protease ER 60, Exp Parasitol 86, 17 (1997)

Gillingham AK and Munro S, The PACT domain, a conserved centrosomal targeting motif in the coiled-coil proteins AKAP450 and pericentrin, EM
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