Applications and advantages of virus-induced gene silencing for gene function studies in plants
Applications and advantages of virus-induced gene silencing for gene function studies in plants
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DOI：
10.1111/j.1365-313x.2004.02158.x
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发表时间：
2004-09-01
期刊：
影响因子：
7.2
通讯作者：
Dinesh-Kumar, SP
中科院分区：
文献类型：
--
作者：
Burch-Smith, TM;Anderson, JC;Dinesh-Kumar, SP
病毒诱导基因沉默（VIGS）是近年来发展起来的一种用于表征植物基因功能的基因转录抑制技术。该方法包括将目标植物基因的短序列克隆到病毒传递载体中。该载体被用来感染幼嫩植物，在几周内，植物抑制病毒复制的自然防御机制也会导致来自内源植物基因的mrna的特异性降解，这些mrna是沉默的目标。VIGS是快速的（从感染到沉默3-4周），不需要发展稳定的转化体，允许表征在稳定品系中可能致命的表型，并提供沉默基因家族单个或多个成员的潜力。在这里，我们简要回顾了导致VIGS发展的发现以及对有效沉默的实验要求的了解。我们描述了VIGS的方法，以及如何优化和用于正向和反向遗传学研究。讨论了与其他植物功能丧失方法相比，VIGS的优点和缺点，以及如何克服VIGS的局限性。本文回顾了VIGS在植物发育和植物防御反应中为研究特定基因的作用提供重要新见解的例子。最后，我们展望了VIGS作为评估和表征植物基因功能的有力工具的未来前景。
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.