Bhardwaj Rita, Krautz-Peterson Greice, Skelly Patrick J
Division of Infectious Diseases, Department of Biomedical Sciences, Cummings School of Veterinary Medicine, Tufts University, North Grafton, MA, USA.
Methods Mol Biol. 2011;764:223-39. doi: 10.1007/978-1-61779-188-8_15.
Schistosomes are parasitic worms that infect over 200 million people and constitute an enormous public health problem worldwide. Molecular tools are being developed for use with these parasites in order to increase our understanding of their unique molecular and cell biology. Among the more promising methodologies is RNA interference (RNAi, or gene silencing), a mechanism by which gene-specific double-stranded RNA (dsRNA) triggers degradation of homologous mRNA transcripts. In this work we describe methods for applying RNAi to suppress gene expression in the intra-mammalian life stages of Schistosoma mansoni. These methods include isolating and culturing the parasites, preparing and delivering dsRNA targeting a specific gene and monitoring the outcome. Given the abundance of schistosome transcriptome and genome sequences now available, RNAi technology has the potential to rapidly expand analysis of the roles and importance of the genes of this globally important parasite.
血吸虫是一种寄生虫,感染人数超过2亿,在全球范围内构成了一个巨大的公共卫生问题。为了增进我们对这些寄生虫独特分子和细胞生物学的了解,正在开发用于研究这些寄生虫的分子工具。在更有前景的方法中,有RNA干扰(RNAi,即基因沉默),这是一种基因特异性双链RNA(dsRNA)触发同源mRNA转录本降解的机制。在这项工作中,我们描述了应用RNAi抑制曼氏血吸虫在哺乳动物体内生命阶段基因表达的方法。这些方法包括分离和培养寄生虫、制备和递送靶向特定基因的dsRNA以及监测结果。鉴于现在已有丰富的血吸虫转录组和基因组序列,RNAi技术有潜力迅速扩展对这种全球重要寄生虫基因的作用和重要性的分析。