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.
目前對(duì)RNAi (RNA interference)的定義有很多種,,不同的資料對(duì)其定義的側(cè)重點(diǎn)也不盡相同,。
如果將RNAi看作一種生物學(xué)現(xiàn)象,可以有以下定義:
① RNAi是由dsRNA介導(dǎo)的由特定酶參與的特異性基因沉默現(xiàn)象,它在轉(zhuǎn)錄水平,、轉(zhuǎn)錄后水平和翻譯水平上阻斷基因的表達(dá),。
② RNAi是有dsRNA參與指導(dǎo)的,,以外源和內(nèi)源mRNA為降解目標(biāo)的轉(zhuǎn)基因沉默現(xiàn)象。具有核苷酸序列特異性的自我防御機(jī)制,,是一種當(dāng)外源基因?qū)牖虿《救肭趾?,?xì)胞中與轉(zhuǎn)基因或入侵病毒RNA同源的基因發(fā)生共同基因沉默的現(xiàn)象。
如果將其作為一門生物技術(shù),,則定義為:
① RNAi 是指通過反義RNA與正鏈RNA 形成雙鏈RNA 特異性地抑制靶基因的現(xiàn)象,它通過人為地引入與內(nèi)源靶基因具有相同序列的雙鏈RNA(有義RNA 和反義RNA) ,從而誘導(dǎo)內(nèi)源靶基因的mRNA 降解,達(dá)到阻止基因表達(dá)的目的,。
② RNAi是指體外人工合成的或體內(nèi)的雙鏈RNA(dsRNA)在細(xì)胞內(nèi)特異性的將與之同源的 mRNA降解成21nt~23nt 的小片段,使相應(yīng)的基因沉默,。
③ RNAi是將與靶基因的mRNA 同源互補(bǔ)的雙鏈RNA(dsRNA ) 導(dǎo)入細(xì)胞,能特異性地降解該mRNA ,從而產(chǎn)生相應(yīng)的功能表型缺失, 屬于轉(zhuǎn)錄后水平的基因沉默(post - transcriptional gene silence , PTGS),。
各種不同定義雖然說法不同,但所描述事實(shí)是大體相同的,,簡(jiǎn)單地可以說,,RNAi就是指由RNA介導(dǎo)的基因沉默現(xiàn)象。
基因沉默:基因沉默是指轉(zhuǎn)基因植物和轉(zhuǎn)基因動(dòng)物中往往會(huì)遇到這樣的情況,,外源基因存在于生物體內(nèi),,并未丟失或損傷,但該基因不表達(dá)或表達(dá)量極低的現(xiàn)象,。
發(fā)生在染色體DNA水平上的轉(zhuǎn)基因沉默叫做位置沉默,;
發(fā)生在RNA轉(zhuǎn)錄水平上的轉(zhuǎn)基因沉默叫做轉(zhuǎn)錄沉默;
發(fā)生在轉(zhuǎn)錄后水平的轉(zhuǎn)基因沉默叫做共抑沉默,。