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Optogenetic control of gene expression in plants in the presence of ambient white light

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DOI

  • Rocio Ochoa-Fernandez, Heinrich Heine University Düsseldorf
  • ,
  • Nikolaj B. Abel
  • Franz Georg Wieland, University of Freiburg
  • ,
  • Jenia Schlegel, Heinrich Heine University Düsseldorf
  • ,
  • Leonie Alexa Koch, Heinrich Heine University Düsseldorf
  • ,
  • J. Benjamin Miller, University of East Anglia
  • ,
  • Raphael Engesser, University of Freiburg
  • ,
  • Giovanni Giuriani, Heinrich Heine University Düsseldorf, University of Glasgow
  • ,
  • Simon M. Brandl, University of Freiburg
  • ,
  • Jens Timmer, University of Freiburg
  • ,
  • Wilfried Weber, University of Freiburg
  • ,
  • Thomas Ott, University of Freiburg
  • ,
  • Rüdiger Simon, Heinrich Heine University Düsseldorf, University of Cologne and Cluster of Excellence on Plant Sciences (CEPLAS)
  • ,
  • Matias D. Zurbriggen, Heinrich Heine University Düsseldorf, University of Cologne and Cluster of Excellence on Plant Sciences (CEPLAS)

Optogenetics is the genetic approach for controlling cellular processes with light. It provides spatiotemporal, quantitative and reversible control over biological signaling and metabolic processes, overcoming limitations of chemically inducible systems. However, optogenetics lags in plant research because ambient light required for growth leads to undesired system activation. We solved this issue by developing plant usable light-switch elements (PULSE), an optogenetic tool for reversibly controlling gene expression in plants under ambient light. PULSE combines a blue-light-regulated repressor with a red-light-inducible switch. Gene expression is only activated under red light and remains inactive under white light or in darkness. Supported by a quantitative mathematical model, we characterized PULSE in protoplasts and achieved high induction rates, and we combined it with CRISPR–Cas9-based technologies to target synthetic signaling and developmental pathways. We applied PULSE to control immune responses in plant leaves and generated Arabidopsis transgenic plants. PULSE opens broad experimental avenues in plant research and biotechnology.

OriginalsprogEngelsk
TidsskriftNature Methods
Vol/bind17
Nummer7
Sider (fra-til)717-725
Antal sider9
ISSN1548-7091
DOI
StatusUdgivet - 1 jul. 2020
Eksternt udgivetJa

Bibliografisk note

Funding Information:
This study was supported in part by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy (CEPLAS—EXC-1028 project no. 194465578 to R.S. and M.D.Z., EXC-2048/1—project no. 390686111 to R.S. and M.D.Z., CIBSS – EXC-2189—project no. 390939984 to T.O., J.T. and W.W., and BIOSS – EXC-294 to J.T. and W.W.), the iGRAD Plant (IRTG 1525 to R.O.F., J.S., R.S. and M.D.Z.), and the Collaborative Research Centers SFB1208 (project no. 267205415; project A13 to M.D.Z.) and SFB924 (INST 95/1126-2; project B4 to T.O.), the European Commission – Research Executive Agency (H2020 Future and Emerging Technologies FET-Open project no. 801041 CyGenTig to M.D.Z.). J.B.M. is supported by a fellowship from the Eastern Academic Research Consortium. We thank D. Orzaez (Polytechnic University of Valencia) and K. Gardner (City University of New York) for kindly providing the GoldenBraid and EL222 plasmids, respectively, T. Brumbarova (University of Düsseldorf) for aid with quantitative reverse-transcription PCR experiments, R. Wurm and M. Gerads (University of Düsseldorf) for technical assistance, and J. Schmidt (Technical Workshop Biology, University of Freiburg) for designing and constructing the light boxes used in this work. We are indebted to J. Casal (University of Buenos Aires), D. Nusinow (Danforth Center), S. Romero, H. Beyer and U. Urquiza (University of Düsseldorf) for careful reading and their suggestions to improve the manuscript.

Publisher Copyright:
© 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

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