Todd Andrea T, Liu Enwu, Page Jonathan E
NRC Plant Biotechnology Institute, Saskatoon, SK, Canada.
Methods Mol Biol. 2010;631:221-36. doi: 10.1007/978-1-60761-646-7_16.
Virus-induced gene silencing (VIGS) exploits endogenous plant antiviral defense mechanisms to posttranscriptionally silence the expression of targeted plant genes. VIGS is quick and relatively easy to perform and therefore serves as a powerful tool for high-throughput functional genomics in plants. Combined with the use of subtractive cDNA libraries for generating a collection of VIGS-ready cDNA inserts, VIGS can be utilized to screen a large number of genes to determine phenotypes resulting from the knockdown/knockout of gene function. Taking into account the optimal insert design for VIGS, we describe a methodology for producing VIGS-ready cDNA libraries enriched for inserts relevant to the biological process of interest.
病毒诱导的基因沉默（VIGS）利用植物内源性抗病毒防御机制在转录后沉默目标植物基因的表达。VIGS快速且相对易于操作，因此是植物高通量功能基因组学的强大工具。结合使用消减cDNA文库来生成一系列适用于VIGS的cDNA插入片段，VIGS可用于筛选大量基因，以确定基因功能敲低/敲除所产生的表型。考虑到VIGS的最佳插入片段设计，我们描述了一种生产适用于VIGS的cDNA文库的方法，该文库富含与感兴趣的生物学过程相关的插入片段。
