Suppression of gene expression by RNA interference in cultured plant cells.
Suppression by double-stranded RNA (dsRNA) of the expression of a target gene is known as RNA interference (RNAi). No quantitative analysis of the effects of RNAi on the expression of specific genes in cultured plant cells has been reported. However, as it is possible to produce populations of cultured plant cells that are uniform and divide synchronously for functional analysis of genes of interest, we performed a quantitative study of the effects of RNAi in such cells. We constructed dsRNA expression plasmids for a luciferase gene under the control of the cauliflower mosaic virus (CaMV) 35S promoter by simply connecting sense and antisense sequences in a head-to-head manner. An RNAi effect was observed 24 hours after the introduction of dsRNA expression plasmids into tobacco BY-2 cells by electroporation. The simple system for suppression of specific genes in plant cells should be useful in attempts to elucidate the roles of individual genes in plant cells.
Akashi
H
H
Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Hongo, Japan.
Miyagishi
M
M
Taira
K
K
D000818:Animals; D002478:Cells, Cultured; D003063:Cnidaria / Q000201:enzymology / Q000235:genetics; D018274:Electroporation; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics; D010957:Plasmids; D034741:RNA, Small Interfering; D022661:RNA, Untranslated / Q000235:genetics*; D013489:Suppression, Genetic; D014026:Nicotiana / Q000235:genetics*; D014162:Transfection
D034741:RNA, Small Interfering; D022661:RNA, Untranslated; D008156:Luciferases