Tobacco rattle virus (TRV)-based virus-induced gene silencing.
Tobacco rattle virus (TRV)-based virus-induced gene silencing.
复制标题
DOI：
10.1007/978-1-61779-882-5_6
复制
发表时间：
2012-01-01
期刊：
影响因子：
--
通讯作者：
Dinesh-Kumar, Savithramma P
中科院分区：
文献类型：
--
作者：
Bachan, Shawn;Dinesh-Kumar, Savithramma P
转录后基因沉默（PTGS）是植物防御病毒最有效的方式之一。这个过程意味着病毒可以触发宿主植物的RNA沉默机制，并最终成为RNA沉默的目标。因此，PTGS是一个有吸引力的内源性过程，可用于研究基因功能。启动PTGS的最有效方法之一是通过病毒诱导的基因沉默（VIGS）。当携带宿主来源的靶基因序列的重组病毒载体（VIGS载体）感染植物时，合成病毒双链RNA，导致抗病毒RNA沉默途径的激活和随后内源宿主基因的敲低。VIGS是一种强大的反向和正向遗传学工具，可用于高通量遗传筛选和功能基因组学。在这里，我们描述了使用烟草脆裂病毒为基础的VIGS技术来研究本氏烟草，番茄和拟南芥的基因功能。
One of the most effective forms of plant defense against viruses is posttranscriptional gene silencing (PTGS). This process implies that a virus can trigger the host plant's RNA-silencing machinery and ultimately become the target of RNA silencing. PTGS is, therefore, an attractive endogenous process that can be exploited to study gene function. One of the most efficient approaches of initiating PTGS is through virus-induced gene silencing (VIGS). When a recombinant viral vector (VIGS vector), carrying a host-derived target gene sequence, infects a plant, viral double-stranded RNAs are synthesized leading to the activation of the antiviral RNA silencing pathway and the subsequent knockdown of the endogenous host gene. VIGS is both a powerful reverse and forward genetics tool and is amendable to high-throughput genetic screens and functional genomics. Here, we describe the use of tobacco rattle virus-based VIGS techniques to study gene function in Nicotiana benthamiana, tomato, and Arabidopsis thaliana.