Doxycycline-dependent photoactivated gene expression in eukaryotic systems

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • S. B. Cambridge - , Heidelberg University , TUD Dresden University of Technology, Max Planck Institute of Neurobiology (Author)
  • D. Geissler - , Max Planck Institute of Neurobiology, Leibniz Institute for Molecular Pharmacology (Author)
  • F. Calegari - , Center for Regenerative Therapies Dresden, Heidelberg University , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • K. Anastassiadis - , Stem Cell Engineering (Junior Research Group), Heidelberg University , BioInnovationsZentrum (Author)
  • M. T. Hasan - , Max Planck Institute of Experimental Medicine (Author)
  • A. F. Stewart - , BioInnovationsZentrum (Author)
  • W. B. Huttner - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • V. Hagen - , Leibniz Institute for Molecular Pharmacology (Author)
  • T. Bonhoeffer - , Max Planck Institute of Neurobiology (Author)

Abstract

High spatial and temporal resolution of conditional gene expression is typically difficult to achieve in whole tissues or organisms. We synthesized two reversibly inhibited, photoactivatable ('caged') doxycycline derivatives with different membrane permeabilities for precise spatial and temporal light-controlled activation of transgenes based on the 'Tet-on' system. After incubation with caged doxycycline or caged cyanodoxycycline, we induced gene expression by local irradiation with UV light or by two-photon uncaging in diverse biological systems, including mouse organotypic brain cultures, developing mouse embryos and Xenopus laevis tadpoles. The amount of UV light needed for induction was harmless as we detected no signs of toxicity. This method allows high-resolution conditional transgene expression at different spatial scales, ranging from single cells to entire complex organisms.

Details

Original languageEnglish
Pages (from-to)527-531
Number of pages5
JournalNature Methods
Volume6
Issue number7
Publication statusPublished - 2009
Peer-reviewedYes

External IDs

Scopus 67649625533
researchoutputwizard legacy.publication#51823
ORCID /0000-0002-4754-1707/work/160953374

Keywords

Keywords

  • Doxycycline-dependent photoactivated gene expression in eukaryotic systems