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          Institute: MPI für Neurobiologie     Collection: Cellular and Systems Neurobiology     Display Documents



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ID: 433289.0, MPI für Neurobiologie / Cellular and Systems Neurobiology
Doxycycline-dependent photoactivated gene expression in eukaryotic systems
Authors:Cambridge, S. B.; Geissler, D.; Calegari, F.; Anastassiadis, K.; Hasan, M. T.; Stewart, A. F.; Huttner, W. B.; Hagen, V.; Bonhoeffer, T.
Language:English
Date of Publication (YYYY-MM-DD):2009-07
Title of Journal:Nature Methods
Journal Abbrev.:Nat. Methods
Volume:6
Issue / Number:7
Start Page:527
End Page:531
Review Status:Peer-review
Audience:Not Specified
Abstract / Description: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.
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Affiliations:MPI für Neurobiologie/Cellular and Systems Neurobiology (Bonhoeffer)
MPI für molekulare Zellbiologie und Genetik
MPI für medizinische Forschung
External Affiliations:[Geissler, Daniel; Hagen, Volker] Leibniz Inst Mol Pharmacol, Berlin, Germany.; [Anastassiadis, Konstantinos; Stewart, A. Francis] Tech Univ Dresden, BioInnovat Zentrum, D-8027 Dresden, Germany.
Identifiers:ISI:000267442900023 [ID No:1]
ISSN:1548-7091 [ID No:2]
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