Sui Guangchao, Soohoo Christina, Affar El Bachir, Gay Frédérique, Shi Yujiang, Forrester William C, Shi Yang
Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115,USA.
Proc Natl Acad Sci U S A. 2002 Apr 16;99(8):5515-20. doi: 10.1073/pnas.082117599.
Double-stranded RNA-mediated interference (RNAi) has recently emerged as a powerful reverse genetic tool to silence gene expression in multiple organisms including plants, Caenorhabditis elegans, and Drosophila. The discovery that synthetic double-stranded, 21-nt small interfering RNA triggers gene-specific silencing in mammalian cells has further expanded the utility of RNAi into mammalian systems. Here we report a technology that allows synthesis of small interfering RNAs from DNA templates in vivo to efficiently inhibit endogenous gene expression. Significantly, we were able to use this approach to demonstrate, in multiple cell lines, robust inhibition of several endogenous genes of diverse functions. These findings highlight the general utility of this DNA vector-based RNAi technology in suppressing gene expression in mammalian cells.
双链RNA介导的干扰（RNAi）最近已成为一种强大的反向遗传学工具，可在包括植物、秀丽隐杆线虫和果蝇在内的多种生物体中使基因表达沉默。合成的双链21核苷酸小干扰RNA能在哺乳动物细胞中触发基因特异性沉默这一发现，进一步拓展了RNAi在哺乳动物系统中的应用。在此，我们报告一种技术，该技术可在体内从DNA模板合成小干扰RNA，以有效抑制内源性基因表达。重要的是，我们能够利用这种方法在多个细胞系中证明对多种功能各异的内源性基因有强力抑制作用。这些发现凸显了这种基于DNA载体的RNAi技术在抑制哺乳动物细胞基因表达方面的普遍实用性。
