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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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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-0EDE-E Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002C-0EE0-6
Genre: Journal Article
Alternative Title : Doxycycline-dependent photoactivated gene expression in eukaryotic systems

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NatMeth_6_2009_527.pdf (Any fulltext), 494KB
 
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Cambridge, Sidney B., Author
Geissler, Daniel, Author
Calegari, Frederico, Author
Anastassiadis, Konstantinos, Author
Hasan, Mazahir T.1, 2, Author              
Stewart, Andrew Francis, Author
Huttner, Wieland B., Author
Hagen, Volker, Author
Bonhoeffer, Tobias, Author
Affiliations:
1Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497704              
2Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497699              

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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: 2009-02-282009-05-152009-06-072009-07
 Publication Status: Published in print
 Pages: 5
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
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Title: Nature methods
  Other : Nature methods
Source Genre: Journal
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Publ. Info: New York, NY : Nature Pub. 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