Induction of stable epigenetic gene silencing in plants using a virus vector.
Gene silencing through transcriptional repression can be induced by double-stranded RNA targeted to a gene promoter, a process known as RNA-mediated transcriptional gene silencing (TGS). This phenomenon is associated with epigenetic changes involving cytosine methylation of the promoter. Plant virus vectors have been used to induce RNA-mediated TGS. Here, we describe methods relevant to the induction of epigenetic changes and RNA-mediated TGS in plants using a virus vector, which include inoculation of recombinant virus, detection of short interfering RNAs, bisulfite sequencing analysis, and nuclear run-on transcription assay.
Research Faculty of Agriculture, Hokkaido University, Kita 9, Nishi 9, Kita-ku, Sapporo, 060-8589, Japan, kanazawa@res.agr.hokudai.ac.jp.
Akira
A
Kanazawa
Megumi
M
Kasai
D019175:DNA Methylation; D020868:Gene Silencing; D005822:Genetic Vectors / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics*; D010942:Plant Viruses / Q000235:genetics*; D011401:Promoter Regions, Genetic; D018749:RNA, Plant / Q000235:genetics; D017423:Sequence Analysis, RNA; D014026:Nicotiana / Q000235:genetics / Q000821:virology*
D010940:Plant Proteins; D018749:RNA, Plant