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Negative regulation of the RTBV promoter by designed zinc finger proteins

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Abstract

The symptoms of rice tungro disease are caused by infection by a DNA-containing virus, rice tungro bacilliform virus (RTBV). To reduce expression of the RTBV promoter, and to ultimately reduce virus replication, we tested three synthetic zinc finger protein transcription factors (ZF-TFs), each comprised of six finger domains, designed to bind to sequences between −58 and +50 of the promoter. Two of these ZF-TFs reduced expression from the promoter in transient assays and in transgenic Arabidopsis thaliana plants. One of the ZF-TFs had significant effects on plant regeneration, apparently as a consequence of binding to multiple sites in the A. thaliana genome. Expression from the RTBV promoter was reduced by ~45% in transient assays and was reduced by up to 80% in transgenic plants. Co-expression of two different ZF-TFs did not further reduce expression of the promoter. These experiments suggest that ZF-TFs may be used to reduce replication of RTBV and thereby offer a potential method for control of an important crop disease.

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Acknowledgments

We thank Dr. Jitender Yadav for his assistance with the RNA work and Dr. Shunhong Dai for providing some materials and for helpful discussions during the early stages of this work. This study was supported by Department of Energy grant DOE-FG02-99ER20355, and NASA grant NNJ04HG98G to RNB and by a grant from The Skaggs Institute for Chemical Biology to CFB.

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  1. Laurent Magnenat

    Present address: Merck Serono S.A., 9 Chemin des Mines, 1202, Geneva, Switzerland

Authors and Affiliations

  1. Donald Danforth Plant Science Center, 975 N. Warson Road, St. Louis, MO, 63132, USA

    M. Isabel Ordiz & Roger N. Beachy

  2. The Skaggs Institute for Chemical Biology and the Department of Molecular Biology, The Scripps Research Institute, BCC-550, North Torrey Pines Road, La Jolla, CA, 92037, USA

    Laurent Magnenat & Carlos F. Barbas III

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  1. M. Isabel Ordiz
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Correspondence to Roger N. Beachy.

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Ordiz, M.I., Magnenat, L., Barbas, C.F. et al. Negative regulation of the RTBV promoter by designed zinc finger proteins. Plant Mol Biol 72, 621–630 (2010). https://doi.org/10.1007/s11103-010-9600-0

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