【摘要】：RNAi 是指同源雙鏈 RNA 所引發(fā)的特異序列的轉(zhuǎn)錄后基因沉默現(xiàn)象。哺乳動物細胞中,小于 30bp 的小雙鏈 RNA 可以避開干擾素反應(yīng)并在哺乳動物細胞內(nèi)引發(fā)基因沉默。在功能基因組研究、藥物靶點篩選、病毒防治、腫瘤治療等研究領(lǐng)域中,RNAi 都是一種非常有用的基因敲除方法,具有廣泛的應(yīng)用前景�，F(xiàn)有的在真核細胞中的 RNAi 技術(shù)可以分為兩大類:⑴ 通過各種方法在體外制備小干涉 RNA(small interfering RNA,siRNA)后直接將其遞送到真核細胞中。 ⑵ 將表達產(chǎn)物在細胞內(nèi)自發(fā)形成 siRNA 的 DNA 表達框遞送到真核細胞中。而所有這些方法的實現(xiàn)都依賴于建立一種核酸遞送系統(tǒng),即一種能夠有效包裹和傳遞外源的遺傳物質(zhì)進入特定類型細胞的高效、低毒的載體系統(tǒng)。 核酸遞送載體系統(tǒng)可以被分為生物載體(主要是指病毒載體)和非生物載體兩大類。病毒載體有著感染細胞的天性,有一整套將自己的基因?qū)氚屑毎牟襟E。然而制備與生產(chǎn)的困難、細胞毒性、免疫原性以及包裝外源基因的大小限制是病毒載體應(yīng)用面對的主要障礙。非病毒類核酸遞送系統(tǒng)往往是在生理條件下帶正電荷的化合物,包括陽離子多聚物,陽離子脂質(zhì)體等。它們通過電荷相互作用與陰性負荷的核酸相互作用并通過內(nèi)吞作用進入到細胞內(nèi)。它不需要面對病毒類載體的種種不利,制備簡便,沒有免疫原性。但是它們導(dǎo)入外源基因的效率遠低于病毒類載體,如何提高效率和穩(wěn)定性,保證安全性一直是研究關(guān)注的重點。
[Abstract]:RNAi refers to posttranscriptional gene silencing of specific sequences induced by homologous double stranded RNA. In mammalian cells, small double-stranded RNA smaller than 30bp can avoid interferon reaction and induce gene silencing in mammalian cells. RNAi is a very useful gene knockout method in the fields of functional genome research, drug target screening, virus prevention and treatment, tumor therapy and so on. The existing RNAi techniques in eukaryotic cells can be divided into two categories: 1: 1 is delivered directly to eukaryotic cells by a variety of methods to prepare small interfering RNA (small interfering RNA,siRNA in vitro. 2 the expressed products are spontaneously formed in the cells The DNA expression box of siRNA was delivered to eukaryotic cells. The realization of all these methods depends on the establishment of a nucleic acid delivery system, that is, a highly efficient and low toxic carrier system that can effectively encapsulate and transmit exogenous genetic material into specific types of cells. Nucleic acid delivery vector systems can be divided into biological carriers (mainly refers to virus vectors) and abiotic carriers. Viral vectors have the nature of infecting cells and a set of steps to transfer their genes to target cells. However, the difficulties of preparation and production, cytotoxicity, immunogenicity and the size limitation of packaging foreign genes are the main obstacles in the application of virus vectors. Non-viral nucleic acid delivery systems are often positive charge compounds under physiological conditions, including cationic polymers, cationic liposomes and so on. They interact with negative-loaded nucleic acids through charge interactions and enter the cell through endocytosis. It does not need to face the various disadvantages of viral vectors, easy to prepare, no immunogenicity. However, the efficiency of introducing foreign genes is much lower than that of virus-like vectors, so how to improve the efficiency and stability and ensure the safety has always been the focus of research.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：Q78

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