Purkayastha Arunima, Sharma Shweta, Dasgupta Indranil
Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi, India.
Methods Mol Biol. 2013;975:33-45. doi: 10.1007/978-1-62703-278-0_3.
Virus-induced gene silencing (VIGS) is a reverse genetics technique that is based on the RNA-mediated defense against viruses in plants. VIGS is a method of gene knockdown triggered by a replicating viral nucleic acid engineered to carry a host gene to be silenced. While there are a number of excellent VIGS vectors available for dicots, only a few are available for monocots. Here, we describe the detailed method of the use of a newly developed VIGS vector for rice, based on the rice-infecting Rice tungro bacilliform virus, a pararetrovirus with dsDNA genome. Using a method based on Agrobacterium-mediated injection of the VIGS construct at the meristematic region of young rice plants, silencing of target genes can be achieved and the silenced phenotype can be visualized in 3 weeks.
病毒诱导的基因沉默（VIGS）是一种反向遗传学技术，它基于植物中RNA介导的抗病毒防御机制。VIGS是一种由经工程改造携带待沉默宿主基因的复制病毒核酸引发的基因敲低方法。虽然有许多优秀的VIGS载体可用于双子叶植物，但可用于单子叶植物的却很少。在此，我们描述了一种基于感染水稻的水稻东格鲁杆状病毒（一种具有双链DNA基因组的类逆转录病毒）新开发的水稻VIGS载体的详细使用方法。使用基于农杆菌介导在水稻幼苗分生组织区域注射VIGS构建体的方法，可以实现靶基因的沉默，并且沉默表型可在3周内显现出来。
