基因沉默
生物
基因
反向遗传学
功能(生物学)
烟草响尾蛇病毒
计算生物学
遗传学
病毒载体
载体(分子生物学)
基因组
重组DNA
作者
Tessa M. Burch‐Smith,Jeffrey C. Anderson,Gregory B. Martin,Savithramma P. Dinesh‐Kumar
出处
期刊:Plant Journal
[Wiley]
日期:2004-08-16
卷期号:39 (5): 734-746
被引量:706
标识
DOI:10.1111/j.1365-313x.2004.02158.x
摘要
Virus-induced gene silencing (VIGS) is a recently developed gene transcript suppression technique for characterizing the function of plant genes. The approach involves cloning a short sequence of a targeted plant gene into a viral delivery vector. The vector is used to infect a young plant, and in a few weeks natural defense mechanisms of the plant directed at suppressing virus replication also result in specific degradation of mRNAs from the endogenous plant gene that is targeted for silencing. VIGS is rapid (3-4 weeks from infection to silencing), does not require development of stable transformants, allows characterization of phenotypes that might be lethal in stable lines, and offers the potential to silence either individual or multiple members of a gene family. Here we briefly review the discoveries that led to the development of VIGS and what is known about the experimental requirements for effective silencing. We describe the methodology of VIGS and how it can be optimized and used for both forward and reverse genetics studies. Advantages and disadvantages of VIGS compared with other loss-of-function approaches available for plants are discussed, along with how the limitations of VIGS might be overcome. Examples are reviewed where VIGS has been used to provide important new insights into the roles of specific genes in plant development and plant defense responses. Finally, we examine the future prospects for VIGS as a powerful tool for assessing and characterizing the function of plant genes.
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