Svoboda Petr, Stein Paula
Institute of Molecular Genetics, Academy of Sciences of Czech Republic, Videnska 1083, 142 20 Prague 4, Czech Republic.
Cold Spring Harb Protoc. 2009 Jan;2009(1):pdb.top56. doi: 10.1101/pdb.top56.
The discovery of RNA interference (RNAi) in 1998 ushered in a new era in biology. RNAi currently serves as a favorite approach for inhibition of gene function in many areas of research. This article provides a brief review of RNAi and discussion of the benefits and drawbacks of using long double-stranded RNA (dsRNA) in mammalian oocytes and early embryos. We also provide an introduction to protocols for RNAi experiments in mouse, including preparation and microinjection of dsRNA into mouse oocytes and early embryos, and preparation and testing of constructs for transgenic RNAi based on long hairpin RNA expression.
1998年RNA干扰(RNAi)的发现开创了生物学的新纪元。目前,RNAi在许多研究领域已成为抑制基因功能的首选方法。本文简要回顾了RNAi,并讨论了在哺乳动物卵母细胞和早期胚胎中使用长双链RNA(dsRNA)的利弊。我们还介绍了小鼠RNAi实验的方案,包括dsRNA的制备及其向小鼠卵母细胞和早期胚胎中的显微注射,以及基于长发夹RNA表达的转基因RNAi构建体的制备和测试。