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  Doxycycline-dependent photoactivated gene expression in eukaryotic systems

Cambridge, S. B., Geissler, D., Calegari, F., Anastassiadis, K., Hasan, M. T., Stewart, A. F., et al. (2009). Doxycycline-dependent photoactivated gene expression in eukaryotic systems. Nature Methods, 6(7), 527-531. doi:10.1038/nmeth.1340.

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Cambridge, S. B.1, Author              
Geissler, D.1, Author              
Calegari, F.2, Author
Anastassiadis, K.3, Author
Hasan, M. T.2, Author
Stewart, A. F.3, Author
Huttner, W. B.2, Author
Hagen, V.3, Author
Bonhoeffer, T.1, Author              
Affiliations:
1Department: Cellular and Systems Neurobiology / Bonhoeffer, MPI of Neurobiology, Max Planck Society, ou_1113545              
2Max Planck Society, ou_persistent13              
3[Geissler, Daniel; Hagen, Volker] Leibniz Inst Mol Pharmacol, Berlin, Germany.; [Anastassiadis, Konstantinos; Stewart, A. Francis] Tech Univ Dresden, BioInnovat Zentrum, D-8027 Dresden, Germany., ou_persistent22              

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 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.

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Language(s): eng - English
 Dates: 2009-07
 Publication Status: Published in print
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 433289
ISI: 000267442900023
DOI: 10.1038/nmeth.1340
 Degree: -

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Title: Nature Methods
  Other : Nature Methods
Source Genre: Journal
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Publ. Info: New York, NY : Nature Publishing Group
Pages: - Volume / Issue: 6 (7) Sequence Number: - Start / End Page: 527 - 531 Identifier: ISSN: 1548-7091
CoNE: https://pure.mpg.de/cone/journals/resource/111088195279556