Open AccessInhibition of gene expression in plant cells by expression of antisense RNA
Author(s) -
Joseph R. Ecker,
Ronald W. Davis
Publication year - 1986
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.83.15.5372
Subject(s) - chloramphenicol acetyltransferase , antisense rna , biology , sense (electronics) , gene , gene expression , rna , electroporation , transcription (linguistics) , acetyltransferase , genetics , rna interference , plasmid , microbiology and biotechnology , promoter , chemistry , linguistics , philosophy , acetylation
Due to the paucity of mutations in biochemical pathways in plants, an alternative approach to classical genetics was tested by expressing antisense RNA in plant cells. A series of plasmids was constructed with the bacterial gene for chloramphenicol acetyltransferase (EC 2.3.1.28) linked in either the sense or antisense orientation to several different plant gene promoters. Various ratios of sense and antisense chloramphenicol acetyltransferase gene plasmids were introduced into plant protoplasts by electric field-mediated DNA transfer (“electroporation”) and the level of expression in each combination was monitored by chloramphenicol acetyltransferase assays. Transcription of antisense RNA was found to effectively block the expression of target genes. Thus, the observation that antisense RNA inhibits gene expression in bacteria and animal systems has been extended to the plant kingdom. Antisense RNA techniques have immediate practical applications in both basic research and in plant genetic engineering.