Journal Article

Regulation of Gene Expression in <i>Arabidopsis thaliana</i> by Artificial Zinc Finger Chimeras

Juan-Pablo Sanchez, Chris Ullman, Michael Moore, Yen Choo and Nam-Hai Chua

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 43, issue 12, pages 1465-1472
Published in print December 2002 | ISSN: 0032-0781
Published online December 2002 | e-ISSN: 1471-9053 | DOI:
Regulation of Gene Expression in Arabidopsis thaliana by Artificial Zinc Finger Chimeras

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


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The artificial regulation of endogenous gene expression in plants is limited to only a few approaches. Here, we describe the use of artificial zinc finger chimeras to regulate the expression of a known reporter construct. The artificial zinc finger chimera TFIIIAZif is a fusion protein consisting of the four zinc fingers of TFIIIA linked through a spacer region to the three zinc fingers of Zif268. This artificial zinc finger chimera is able to bind specifically to a target DNA sequence (ZBS, zinc finger binding site) of 27 base pairs (bp). TFIIIAZif was fused to a transactivation domain from the herpes simplex virus VP16 or its tetramer VP64 to give ZF-VP16 or ZF-VP64, respectively. In transient expression assays, these two transcription activators were able to activate a target reporter gene (luc and GFP) expressed from a minimal –46 35S promoter linked to four copies of ZBS. The activation was confirmed in transgenic plants using an inducible XVE system [Zuo et al. (2000) Plant J. 24: 265] to express ZF-VP16 or ZF-VP64. Furthermore, to test the specificity of ZF-VP64 we have compared reporter gene expression from a wild type (1×ZBS) and a mutant (1×ZBSmu) binding site in transgenic plants. The 1×ZBS was used to express green fluorescent protein (GFP) whereas the 1×ZBSmu was used to express red fluorescent protein (RFP). Upon induction of ZF-VP64 we found a much higher expression of GFP (about 33-fold) as compared to RFP expression. These results suggest that artificial zinc finger chimeras can be used to target specific DNA sequences and to regulate gene expression in plants.

Keywords: Keywords: Arabidopsis —Gene expression — Regulation — Zinc finger.; Abbreviations: ACT, actin; CaMV, cauliflower mosaic virus; GFP, green fluorescent protein; Luc, luciferase; NLS, nuclear localization signal; RFP, red fluorescent protein; VP16, transactivation domain from the herpes simplex virus; VP64, a tetramer of VP16; XVE, estrogen receptor-based inducible system; ZBS, zinc finger chimera binding site; ZBSmu, zinc finger chimera binding site with a mutation; ZF, zinc finger chimera.

Journal Article.  5455 words.  Illustrated.

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

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