Institute for Parasitology, Justus-Liebig-University, 35392 Giessen, Germany.
Parasitology. 2012 Apr;139(5):651-68. doi: 10.1017/S0031182011001466. Epub 2011 Sep 6.
In parasitological research, significant progress has been made with respect to genomics and transcriptomics but transgenic systems for functional gene analyses are mainly restricted to the protozoan field. Gene insertion and knockout strategies can be applied to parasitic protozoa as well as gene silencing by RNA interference (RNAi). By contrast, research on parasitic helminthes still lags behind. Along with the major advances in genome and transcriptome analyses e.g. for schistosomes, methods for the functional characterization of genes of interest are still in their initial phase and have to be elaborated now, at the beginning of the post-genomic era. In this review we will summarize attempts made in the last decade regarding the establishment of protocols to transiently and stably transform or transfect schistosomes. Besides approaches using particle bombardment, electroporation or virus-based infection strategies to introduce DNA constructs into adult and larval schistosome stages to express reporter genes, first approaches have also been made in establishing protocols based on soaking, lipofection, and/or electroporation for RNA interference to silence gene activity. Although in these cases remarkable progress can be seen, the schistosome community eagerly awaits major breakthroughs especially with respect to stable transformation, but also for silencing or knock-down strategies for every schistosome gene of interest.
在寄生虫学研究中,基因组学和转录组学取得了重大进展,但功能基因分析的转基因系统主要局限于原生动物领域。基因插入和敲除策略可应用于寄生原生动物,也可通过 RNA 干扰 (RNAi) 进行基因沉默。相比之下,寄生虫蠕虫的研究仍然滞后。随着基因组和转录组分析的重大进展,例如对血吸虫的分析,对感兴趣基因的功能特征进行分析的方法仍处于初始阶段,现在必须在基因组时代的初期进行阐述。在这篇综述中,我们将总结过去十年中在建立瞬时和稳定转化或转染血吸虫的方案方面所做的尝试。除了使用粒子轰击、电穿孔或基于病毒的感染策略将 DNA 构建体引入成虫和幼虫血吸虫阶段以表达报告基因之外,还首次尝试了基于浸泡、脂质转染和/或电穿孔的方案,以进行 RNAi 沉默基因活性。尽管在这些情况下可以看到显著的进展,但血吸虫社区急切地等待着重大突破,特别是在稳定转化方面,但也希望针对每个感兴趣的血吸虫基因,实现沉默或敲低策略。