title: Photoactivation of the CreER T2 recombinase for conditional site-specific recombination with high spatiotemporal resolution.
abstract: We implemented a noninvasive optical method for the fast control of Cre recombinase in single cells of a live zebrafish embryo. Optical uncaging of the caged precursor of a nonendogeneous steroid by one- or two-photon illumination was used to restore Cre activity of the CreER(T2) fusion protein in specific target cells. This method labels single cells irreversibly by inducing recombination in an appropriate reporter transgenic animal and thereby can achieve high spatiotemporal resolution in the control of gene expression. This technique could be used more generally to investigate important physiological processes (e.g., in embryogenesis, organ regeneration, or carcinogenesis) with high spatiotemporal resolution (single cell and 10-min scales).
authors.affiliation: Ecole Normale Supérieure, Laboratoire de Physique Statistique, UMR 8550 CNRS, Paris Cedex, France.
authors.forename: Deepak Kumar
authors.initials: DK
authors.lastname: Sinha
authors.forename: Pierre
authors.initials: P
authors.lastname: Neveu
authors.forename: Nathalie
authors.initials: N
authors.lastname: Gagey
authors.forename: Isabelle
authors.initials: I
authors.lastname: Aujard
authors.forename: Thomas
authors.initials: T
authors.lastname: Le Saux
authors.forename: Christine
authors.initials: C
authors.lastname: Rampon
authors.forename: Carole
authors.initials: C
authors.lastname: Gauron
authors.forename: Koichi
authors.initials: K
authors.lastname: Kawakami
authors.forename: Christoph
authors.initials: C
authors.lastname: Leucht
authors.forename: Laure
authors.initials: L
authors.lastname: Bally-Cuif
authors.forename: Michel
authors.initials: M
authors.lastname: Volovitch
authors.forename: David
authors.initials: D
authors.lastname: Bensimon
authors.forename: Ludovic
authors.initials: L
authors.lastname: Jullien
authors.forename: Sophie
authors.initials: S
authors.lastname: Vriz
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D017931:DNA Primers / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D019426:Integrases / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D055668:Photochemical Processes; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D011995:Recombination, Genetic / Q000502:physiology*; D013050:Spectrometry, Fluorescence; D015027:Zebrafish
chemical_list: D017931:DNA Primers; D018840:HSP70 Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; C045073:Cre recombinase; D019426:Integrases