Lenz G
Departamento de Biofísica, Federal do Rio Grande do Sul, Av. Benito Goncalves 9500, 91501-370 Porto Alegre, RS, Brasil.
Braz J Med Biol Res. 2005 Dec;38(12):1749-57. doi: 10.1590/s0100-879x2005001200003. Epub 2005 Nov 9.
The discovery of double-stranded RNA-mediated gene silencing has rapidly led to its use as a method of choice for blocking a gene, and has turned it into one of the most discussed topics in cell biology. Although still in its infancy, the field of RNA interference has already produced a vast array of results, mainly in Caenorhabditis elegans, but recently also in mammalian systems. Micro-RNAs are short hairpins of RNA capable of blocking translation, which are transcribed from genomic DNA and are implicated in several aspects from development to cell signaling. The present review discusses the main methods used for gene silencing in cell culture and animal models, including the selection of target sequences, delivery methods and strategies for a successful silencing. Expected developments are briefly discussed, ranging from reverse genetics to therapeutics. Thus, the development of the new paradigm of RNA-mediated gene silencing has produced two important advances: knowledge of a basic cellular mechanism present in the majority of eukaryotic cells and access to a potent and specific new method for gene silencing.
双链RNA介导的基因沉默的发现迅速使其成为阻断基因的首选方法,并使其成为细胞生物学中讨论最多的话题之一。尽管RNA干扰领域仍处于起步阶段,但已经产生了大量研究成果,主要集中在秀丽隐杆线虫中,不过最近在哺乳动物系统中也有相关成果。微小RNA是能够阻断翻译的短RNA发夹结构,它们由基因组DNA转录而来,涉及从发育到细胞信号传导等多个方面。本综述讨论了细胞培养和动物模型中用于基因沉默的主要方法,包括靶序列的选择、递送方法以及成功实现沉默的策略。还简要讨论了预期的发展,从反向遗传学到治疗学。因此,RNA介导的基因沉默这一新范式的发展带来了两个重要进展:对大多数真核细胞中存在的基本细胞机制的认识,以及获得一种有效且特异的基因沉默新方法。