Applications and advantages of virus-induced gene silencing for gene function studies in plants

被引:619
作者
Burch-Smith, TM
Anderson, JC
Martin, GB
Dinesh-Kumar, SP [1 ]
机构
[1] Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA
[2] Cornell Univ, Dept Mol Biol & Genet, Ithaca, NY 14853 USA
[3] Cornell Univ, Boyce Thompson Inst Plant Res, Ithaca, NY 14853 USA
[4] Cornell Univ, Dept Plant Pathol, Ithaca, NY 14853 USA
关键词
virus-induced gene silencing; functional genomics; methods; silencing; TRV-VIGS vector;
D O I
10.1111/j.1365-313X.2004.02158.x
中图分类号
Q94 [植物学];
学科分类号
071001 ;
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.
引用
收藏
页码:734 / 746
页数:13
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