( nsformation protocols for parasitic hel...)Biolistic Gene Transfer to Generate Transgenic Schistosomes, Rev A

nsformation protocols for parasitic helminths have not been established or reproducibly applied. Recently, the first experiments were reported to introduce DNA or RNA into the parasitic nematode, Ascaris lumbricoides, by particle bombardment (Davis et al. 1999). In the same study, the first evidence was obtained that this method could also be applicable to adult schistosomes. However, no molecular results on transgene expression were presented, and no microscopic analyses were performed.

The results demonstrated here show that biolistic gene transfer can be used to transiently transform adult schistosomes as well as larval stages. In our study, two different systems for biolistic gene transfer were used. Results with the Helios gene gun showed that this approach was practicable, but has disadvantages. The rate of worm survival was low and the conditions during bombardment were unfavorable. Worms that survived the procedure showed significant tissue damage. The Helios gene gun has been used successfully to transform the nematode, Caenorhabditis elegans, and the filarial parasite, Litomosoides sigmodontis (Jackstadt et al. 1999). In contrast to schistosomes, filariae are bigger and seem to be more resistant to mechanical stress.

Results obtained with the PDS1000/He system demonstrate that this system is more suitable than the Helios gene gun to transiently transform S. mansoni. In spite of the vacuum, nearly all worms survived the procedure without obvious damage. Likewise, transformation of larval stages was achieved using lower pressures. Molecular analyses showed that the plasmid vector was successfully introduced into both developmental stages; the transgene was transcribed and translated. Microscopic analyses dem
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