Pulukkunat Dileep K, Raj M L Stephen, Pattanayak Debasis, Lai Lien B, Gopalan Venkat
Department of Biochemistry, The Ohio State University, Columbus, OH, USA.
Methods Mol Biol. 2003;236:295-310. doi: 10.1385/1-59259-413-1:295.
As we trek into the uncharted territories of the genomic era, there is an urgency for the development of approaches for assigning functions to the multitude of uncharacterized genes. Although currently available knock-out methodologies could be used for uncovering the function of newly discovered genes, the mixed outcomes in terms of the success of these approaches in down-regulating gene expression necessitate the development of new functional genomics tools. This chapter describes in detail the experimental method for targeted mRNA degradation inside plant cells by enticing the endogenous and ubiquitous RNase P into recognition of specific mRNAs as non-natural substrates.
随着我们踏入基因组时代的未知领域,迫切需要开发方法来为众多未表征基因赋予功能。尽管目前可用的基因敲除方法可用于揭示新发现基因的功能,但这些方法在下调基因表达方面的成功率参差不齐,因此有必要开发新的功能基因组学工具。本章详细描述了一种实验方法,即通过诱使内源性且普遍存在的核糖核酸酶P将特定mRNA识别为非天然底物,从而在植物细胞内实现靶向mRNA降解。