Journal Article

Inducible Antisense-mediated Post-transcriptional Gene Silencing in Transgenic Pine Cells Using Green Fluorescent Protein as a Visual Marker

Wei Tang, Katherine Kinken and Ronald J. Newton

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 46, issue 8, pages 1255-1263
Published in print August 2005 | ISSN: 0032-0781
Published online August 2005 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pci134
Inducible Antisense-mediated Post-transcriptional Gene Silencing in Transgenic Pine Cells Using Green Fluorescent Protein as a Visual Marker

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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An inducible post-transcriptional gene silencing (PTGS) system was established in Virginia pine (Pinus virginiana Mill.) cells. This system is based on the activation of an antisense gfp gene construct by a chimeric transcriptional activator GVG (Gal4-binding domain–VP16 activation domain–glucocorticoid receptor fusion) upon application of the inducer to gfp transgenic cell lines. A detailed characterization of the inducible PTGS system in transgenic cell lines demonstrated that this system is stringently controlled. The degree of silencing with this construct could be regulated by the concentration of inducer and the time of treatment. Such transgenic cell lines may provide a useful system to study signaling mechanisms of gene silencing in transgenic pine cells. The inducible system could be a useful tool for functional discovery of novel plant genes.

Keywords: Chimeric transcription factor; Conditional gene expression; Dexamethasone; Post-transcriptional gene silencing; RNA interference; Transgene; CaMV, cauliflower mosaic virus; DIG, digoxigenin; dsRNA, double-stranded RNA; GFP, green fluorescent protein (gfp, gene); GVG, Gal4-binding domain–VP16 activation domain–glucocorticoid receptor fusion; PTGS, post-transcriptional gene silencing; RdRP, RNA-dependent RNA polymerase; siRNA, small interfering RNA

Journal Article.  6654 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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