title: Doxycycline-dependent photoactivated gene expression in eukaryotic systems.
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.
authors.affiliation: Max Planck Institute of Neurobiology, Munich-Martinsried, Germany. cambridge@ana.uni-heidelberg.de
authors.forename: Sidney B
authors.initials: SB
authors.lastname: Cambridge
authors.forename: Daniel
authors.initials: D
authors.lastname: Geissler
authors.forename: Federico
authors.initials: F
authors.lastname: Calegari
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Anastassiadis
authors.forename: Mazahir T
authors.initials: MT
authors.lastname: Hasan
authors.forename: A Francis
authors.initials: AF
authors.lastname: Stewart
authors.forename: Wieland B
authors.initials: WB
authors.lastname: Huttner
authors.forename: Volker
authors.initials: V
authors.lastname: Hagen
authors.forename: Tobias
authors.initials: T
authors.lastname: Bonhoeffer
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004318:Doxycycline / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D005260:Female; D015870:Gene Expression / Q000187:drug effects* / Q000528:radiation effects*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects; D007814:Larva / Q000187:drug effects / Q000235:genetics / Q000528:radiation effects; D051379:Mice; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton; D018462:Photobiology; D011247:Pregnancy; D011994:Recombinant Proteins / Q000235:genetics; D046509:Tissue Culture Techniques; D014466:Ultraviolet Rays; D014982:Xenopus laevis / Q000235:genetics / Q000254:growth & development
chemical_list: D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D004318:Doxycycline