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siRNA,shRNA,dsRNA,miRNA間的區別
發布日期:2015-03-15 14:50:21 瀏覽次數 :
導讀
RNA干擾(RNAi),即用20多個核苷 酸組成的短的雙鏈RNA(siRNA)代替傳統反義核酸進行轉錄后基因沉默,已經迅速而廣泛地應用到基因功能,基因表達調控機制研究等熱門領域。

siRNA,shRNA,dsRNA,miRNA間的區別

RNA干擾(RNAi),即用20多個核苷 酸組成的短的雙鏈RNA(siRNA)代替傳統反義核酸進行轉錄后基因沉默,已經迅速而廣泛地應用到基因功能,基因表達調控機制研究等熱門領域,并為基因 治療開辟了新的途徑。近兩年來,這方面的科學論文及報道爆炸性增長,幾乎每天都有新的結果涌現。這些論文中,出現了很多與RNA干擾相關的概念,意思很相 近但并不完全相同,這里列舉了幾個常見的

RNA干擾相關的名詞及其簡單的解釋:

1、 miRNA:在遺傳學中,微 RNA(microRNAs,miRNA)是長度在21-23個核苷酸之間的單鏈RNA片段, 調節基因的表達。miRNA由基因編碼,從DNA轉錄而來, 但不翻譯成蛋白。初級轉錄產物(primary transcripts,pri-miRNA)縮短其頸環結構得到功能性的miRNA。成熟的miRNA分子與一個或更多個mRNA分子部分互補,其主要 功能是下調基因的表達。而microRNA這個概念第一次在2001年Science (26 October 2001)的文章中出現。

2、 dsRNA: 雙鏈RNA(double-stranded RNA, dsRNA)是一種有互補鏈的RNA,與細胞中發現的DNA相似,dsRNA構成了一些病毒(雙鏈RNA病毒)的基因組。像病毒RNA或siRNA之類的 雙鏈RNA能夠促發真核細胞中的RNA干擾,引起脊椎動物中的干擾素反應。

3、 shRNA:小發卡或短發卡RNA(a small hairpin RNA or short hairpin RNA, shRNA)是一段具有緊密發卡環(tight hairpin turn)的RNA序列,常被用于RNA干擾沉默靶基因的表達。利用載體把shRNA導入細胞,載體中的U6 啟動子確保shRNA總是表達;這種裝載了shRNA載體可被傳遞到子代細胞中去,從而使基因的沉默可被遺傳。shRNA的發卡結構可被細胞機制切割成 siRNA,然后siRNA結合到RNA誘導沉默復合物上(RNA-induced silencing complex,RISC),該復合物能夠結合到目的mRNAs并將其降解。shRNA 是由RNA 聚合酶Ⅲ轉錄的,哺乳細胞中的shRNA產物有時會促使細胞產生干擾素反應(細胞遇到病毒侵襲時做出的自我防衛反應),而miRNA中卻不會遇到這種問題 (miRNA是由RNA聚合酶Ⅱ轉錄,與轉錄mRNA的聚合酶相同)。shRNA也被用于植物和其它系統中,在這些系統中不需要U6啟動子的驅動。在植物 中,常用的增強表達的啟動子是CaMV35S(
cauliflower mosaic virus 35S),在這種情況下,RNA聚合酶Ⅱ參與了轉錄,起始RNA干擾。

4、 siRNA:小或短干擾RNA(small/short interfering RNA, siRNA)是一類20-25個核苷酸長度的雙鏈RNA分子,其主要在RNAi通路中起作用,干擾特異基因的表達。此外siRNA在RNAi相關的通路中 也起作用,如抗病毒機制,基因組染色體結構的塑造等。其結構是一段完全互補的RNA雙鏈,兩端各有兩個未互補的的堿基。
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