RNA interference (RNAi) by double stranded RNA (dsRNAs) molecules of approximately 20-25 nucleotides termed short interfering (siRNAs) is a powerful method for preventing the expression of a particular gene. The dsRNA dominantly silences gene expression in a sequence-specific manner by causing the corresponding endogenous mRNA to be degraded. The technique was first developed in Caenorhabditis elegans, and was rapidly applied to a wide range of organisms. Methods for expressing siRNAs in cells in culture and in vivo using viral vectors, and for transfecting cells with synthetic siRNAs, have been developed and are being used to establish the functions of specific proteins in various cell types and organisms. For example, chemically synthesized or in vitro transcribed siRNAs can be transfected into cells, injected into mice, or introduced into plants. siRNAs can also be expressed endogenously from siRNA expression vectors or PCR products in cells or in transgenic animals.
目前对RNAi (RNA interference)的定义有很多种,不同的资料对其定义的侧重点也不尽相同。
如果将RNAi看作一种生物学现象,可以有以下定义:
① RNAi是由dsRNA介导的由特定酶参与的特异性基因沉默现象,它在转录水平、转录后水平和翻译水平上阻断基因的表达。
② RNAi是有dsRNA参与指导的,以外源和内源mRNA为降解目标的转基因沉默现象。具有核苷酸序列特异性的自我防御机制,是一种当外源基因导入或病毒入侵后,细胞中与转基因或入侵病毒RNA同源的基因发生共同基因沉默的现象。
如果将其作为一门生物技术,则定义为:
① RNAi 是指通过反义RNA与正链RNA 形成双链RNA 特异性地抑制靶基因的现象,它通过人为地引入与内源靶基因具有相同序列的双链RNA(有义RNA 和反义RNA) ,从而诱导内源靶基因的mRNA 降解,达到阻止基因表达的目的。
② RNAi是指体外人工合成的或体内的双链RNA(dsRNA)在细胞内特异性的将与之同源的 mRNA降解成21nt~23nt 的小片段,使相应的基因沉默。
③ RNAi是将与靶基因的mRNA 同源互补的双链RNA(dsRNA ) 导入细胞,能特异性地降解该mRNA ,从而产生相应的功能表型缺失, 属于转录后水平的基因沉默(post - transcriptional gene silence , PTGS)。
各种不同定义虽然说法不同,但所描述事实是大体相同的,简单地可以说,RNAi就是指由RNA介导的基因沉默现象。
基因沉默:基因沉默是指转基因植物和转基因动物中往往会遇到这样的情况,外源基因存在于生物体内,并未丢失或损伤,但该基因不表达或表达量极低的现象。
发生在染色体DNA水平上的转基因沉默叫做位置沉默;
发生在RNA转录水平上的转基因沉默叫做转录沉默;
发生在转录后水平的转基因沉默叫做共抑沉默。