Wei Ling, Liu Xuan, Cao Cheng
Beying Institute of Biotechnology, Academy of Military Medical Sciences, Beijing 100850, China.
Sheng Wu Gong Cheng Xue Bao. 2005 Jul;21(4):516-9.
RNA interfering (RNAi)--one of the most exciting discoveries in biology in the last couple decades has quickly become one of the most powerful and indispensable tools in the molecular biologist's toolkit. It is an important protection mechanism in cells, by which animals and plants defend viral infection and inhibit viral replication. RNAi is the process of sequence-specific, posttranscriptional gene silencing in animals and plants initiated by dsRNA that is homologous to the silenced gene and has emerged as a powerful tool to silence gene expression in multiple organisms. It has recently been shown that double-stranded, small interfering RNAs (siRNAs) of 21-25 nucleotides can be transfected into relevant cells to target specific RNAs. In mammalian cells it is known that dsRNA 30 base pairs or longer can trigger interferon responses that are intrinsically sequence-nonspecific, thus limiting the application of RNAi as an experimental and therapeutic agent. Duplexes of 19-25 nts (nucleotides) siRNA with short 3' overhangs, however, can efficiently inhibit gene expression in a sequence-specific manner in cultured mammalian cells. This approach was utilized to inhibit human immunodeficiency virus type I (HIV-1) infection in human cells. The in vitro experiments indicate that RNAi as a powerful antiviral tool will bring a promising future to gene therapy for virus disease.
RNA干扰(RNAi)——过去几十年生物学领域最令人兴奋的发现之一,迅速成为分子生物学家工具包中最强大且不可或缺的工具之一。它是细胞中的一种重要保护机制,动植物借此抵御病毒感染并抑制病毒复制。RNAi是动植物中由与沉默基因同源的双链RNA引发的序列特异性、转录后基因沉默过程,已成为在多种生物体中使基因表达沉默的强大工具。最近研究表明,21至25个核苷酸的双链小干扰RNA(siRNA)可转染到相关细胞中以靶向特定RNA。在哺乳动物细胞中,已知30个碱基对或更长的双链RNA可触发本质上序列非特异性的干扰素反应,从而限制了RNAi作为实验和治疗剂的应用。然而,具有短3'突出端的19至25个核苷酸(nts)的siRNA双链体能够以序列特异性方式有效抑制培养的哺乳动物细胞中的基因表达。该方法被用于抑制人类细胞中的I型人类免疫缺陷病毒(HIV-1)感染。体外实验表明,RNAi作为一种强大的抗病毒工具将为病毒疾病的基因治疗带来光明前景。