A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference
A lentivirus-based system to functionally silence genes in primary mammalian cells, stem cells and transgenic mice by RNA interference
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DOI：
10.1038/ng1117
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发表时间：
2003-03-01
期刊：
影响因子：
30.8
通讯作者：
Van Parijs, L
中科院分区：
文献类型：
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作者：
Rubinson, DA;Dillon, CP;Van Parijs, L
RNA干扰（RNAi）最近已成为一种特异且高效的方法，可通过转染短干扰RNA（siRNAs；参考文献1），或者更近一些，通过从表达载体和逆转录病毒转录短发夹RNA（shRNAs）来沉默哺乳动物细胞中的基因表达（2 - 10）。但是重要细胞类型在体外和体内对这些方法转导的抗性（11）限制了RNAi的应用。在此我们描述一种慢病毒系统，用于将shRNAs递送到有丝分裂活跃和不活跃的哺乳动物细胞、干细胞、受精卵及其分化后代中。我们表明慢病毒递送的shRNAs能够在多种细胞类型以及转基因小鼠中对基因表达进行特异、高度稳定且有功能的沉默。我们的慢病毒载体应能允许对原代人类和动物细胞及组织中的基因功能进行快速有效的分析，并产生特定基因表达降低的动物。它们还可能为基因治疗提供新的途径。
RNA interference (RNAi) has recently emerged as a specific and efficient method to silence gene expression in mammalian cells either by transfection of short interfering RNAs (siRNAs; ref. 1) or, more recently, by transcription of short hairpin RNAs (shRNAs) from expression vectors and retroviruses(2-10). But the resistance of important cell types to transduction by these approaches, both in vitro and in vivo(11), has limited the use of RNAi. Here we describe a lentiviral system for delivery of shRNAs into cycling and non-cycling mammalian cells, stem cells, zygotes and their differentiated progeny. We show that lentivirus-delivered shRNAs are capable of specific, highly stable and functional silencing of gene expression in a variety of cell types and also in transgenic mice. Our lentiviral vectors should permit rapid and efficient analysis of gene function in primary human and animal cells and tissues and generation of animals that show reduced expression of specific genes. They may also provide new approaches for gene therapy.