Bellofatto Vivian, Palenchar Jennifer B
Department of Microbiology and Molecular Genetics, UMDNJ-NJ Medical School, International Center for Public Health, Newark, NJ, USA.
Methods Mol Biol. 2008;442:83-94. doi: 10.1007/978-1-59745-191-8_7.
RNA interference (RNAi) is a cellular mechanism that is often exploited as a technique for quelling the expression of a specific gene. RNAi studies are carried out in vivo, making this a powerful means for the study of protein function in situ Several trypanosomatids, including those organisms responsible for human and animal diseases, naturally possess the machinery necessary for RNAi manipulations. This allows for the use of RNAi in unraveling many of the pressing questions regarding the parasite's unique biology. The completion of the Trypanosoma brucei genome sequence, coupled with several powerful genetic tools, has resulted in widespread utilization of RNAi in this organism. The key steps for RNAi-based reduction of gene expression, including parasite cell culture, DNA transfection, RNAi expression, and experimental execution, are discussed with a focus on procyclic forms of Trypanosoma brucei.
RNA干扰（RNAi）是一种细胞机制，常被用作抑制特定基因表达的技术。RNAi研究在体内进行，这使其成为原位研究蛋白质功能的有力手段。包括那些导致人类和动物疾病的生物在内的几种锥虫天然拥有RNAi操作所需的机制。这使得RNAi可用于解答许多有关寄生虫独特生物学的紧迫问题。布氏锥虫基因组序列的完成，再加上几种强大的遗传工具，使得RNAi在这种生物体中得到了广泛应用。本文讨论了基于RNAi降低基因表达的关键步骤，包括寄生虫细胞培养、DNA转染、RNAi表达和实验实施，重点是布氏锥虫的前循环形式。
