Culture for genetic manipulation of developmental stages of Schistosoma mansoni.
Culture for genetic manipulation of developmental stages of Schistosoma mansoni.
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DOI：
10.1017/s0031182009991211
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发表时间：
2010-03
期刊：
影响因子：
2.4
通讯作者：
Brindley PJ
中科院分区：
文献类型：
--
作者：
Mann VH;Morales ME;Rinaldi G;Brindley PJ
主要的人类蠕虫寄生虫的基因组，实际上还有许多其他的农业意义，现在是密集的基因组测序和注释的研究重点。2007年，马来丝虫的基因组序列草图被报道，两种人类寄生虫日本血吸虫和日本血吸虫的基因组草图也被报道。mansoni在2009年报道。这些基因组数据提供了一个全面了解的分子机制，涉及到寄生虫的营养和代谢，宿主依赖的发展和成熟，免疫逃避和无脊椎动物进化的基础。此外，可能会预测新的潜在候选疫苗和药物靶点。然而，由于维持发育周期的困难和费用，测试这些预测通常并不简单。为了促进这一目标，几个发育阶段可以在体外维持更短或更长的时间间隔，并且这些阶段易于操作。我们的研究兴趣集中在血吸虫基因功能的实验研究上，最近重点关注转基因和RNA干扰的发展，以实现可遗传基因操作的长期目标。在这里，我们审查的方法，分离和培养的发展阶段的曼氏血吸虫，包括卵，孢子囊，囊泡和成人，特别是因为这些程序涉及到基因操作的方法。我们还讨论了最近的进展，包括部署的方波电穿孔引入报告基因到培养的染色体的基因操作。
Genomes of the major human helminth parasites, and indeed many others of agricultural significance, are now the research focus of intensive genome sequencing and annotation. A draft genome sequence of the filarial parasite Brugia malayi was reported in 2007 and draft genomes of two of the human schistosomes, Schistosoma japonicum and S. mansoni reported in 2009. These genome data provide the basis for a comprehensive understanding of the molecular mechanisms involved in schistosome nutrition and metabolism, host-dependent development and maturation, immune evasion and invertebrate evolution. In addition, new potential vaccine candidates and drug targets will likely be predicted. However, testing these predictions is often not straightforward with schistosomes because of the difficulty and expense in maintenance of the developmental cycle. To facilitate this goal, several developmental stages can be maintained in vitro for shorter or longer intervals of time, and these are amenable to manipulation. Our research interests focus on experimental studies of schistosome gene functions, and more recently have focused on development of transgenesis and RNA interference with the longer term aim of heritable gene manipulation. Here we review methods to isolate and culture developmental stages of Schistosoma mansoni, including eggs, sporocysts, schistosomules and adults, in particular as these procedures relate to approaches for gene manipulation. We also discuss recent advances in genetic manipulation of schistosomes including the deployment of square wave electroporation to introduce reporter genes into cultured schistosomes.