Department of Microbiology, Immunology and Tropical Medicine, George Washington University Medical Center, Washington, DC, USA.
Mem Inst Oswaldo Cruz. 2011 Nov;106(7):785-93. doi: 10.1590/s0074-02762011000700002.
Genome sequences for Schistosoma japonicum and Schistosoma mansoni are now available. The schistosome genome encodes ~13,000 protein encoding genes for which the function of only a minority is understood. There is a valuable role for transgenesis in functional genomic investigations of these new schistosome gene sequences. In gain-of-function approaches, transgenesis can lead to integration of transgenes into the schistosome genome which can facilitate insertional mutagenesis screens. By contrast, transgene driven, vector-based RNA interference (RNAi) offers powerful loss-of-function manipulations. Our laboratory has focused on development of tools to facilitate schistosome transgenesis. We have investigated the utility of retroviruses and transposons to transduce schistosomes. Vesicular stomatitis virus glycoprotein (VSVG) pseudotyped murine leukemia virus (MLV) can transduce developmental stages of S. mansoni including eggs. We have also observed that the piggyBac transposon is transpositionally active in schistosomes. Approaches with both VSVG-MLV and piggyBac have resulted in somatic transgenesis and have lead to integration of active reporter transgenes into schistosome chromosomes. These findings provided the first reports of integration of reporter transgenes into schistosome chromosomes. Experience with these systems is reviewed herewith, along with findings with transgene mediated RNAi and germ line transgenesis, in addition to pioneering and earlier reports of gene manipulation for schistosomes.
日本血吸虫和曼氏血吸虫的基因组序列现已公布。血吸虫基因组编码了约 13000 个蛋白质编码基因,其中只有少数基因的功能被了解。转基因技术在这些新的血吸虫基因序列的功能基因组研究中具有重要作用。在功能获得方法中,转基因可以整合到血吸虫基因组中,从而促进插入诱变筛选。相比之下,转基因驱动的基于载体的 RNA 干扰 (RNAi) 提供了强大的功能丧失操作。我们的实验室专注于开发工具来促进血吸虫的转基因技术。我们已经研究了逆转录病毒和转座子在转导血吸虫中的应用。水疱性口炎病毒糖蛋白 (VSVG) 假型化的鼠白血病病毒 (MLV) 可以转导曼氏血吸虫的发育阶段,包括卵。我们还观察到,猪囊尾蚴转座子在血吸虫中转座是活跃的。VSVG-MLV 和 piggyBac 的方法都导致了体转基因,并导致活性报告基因整合到血吸虫染色体中。这些发现首次报道了报告基因整合到血吸虫染色体中。本文回顾了这些系统的经验,以及转基因介导的 RNAi 和种系转基因的发现,此外还介绍了早期对血吸虫进行基因操作的先驱性和早期报告。