Photoactivation of the CreER T2 recombinase for conditional site-specific recombination with high spatiotemporal resolution.
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).
Sinha
Deepak Kumar
DK
Ecole Normale Supérieure, Laboratoire de Physique Statistique, UMR 8550 CNRS, Paris Cedex, France.
Neveu
Pierre
P
Gagey
Nathalie
N
Aujard
Isabelle
I
Le Saux
Thomas
T
Rampon
Christine
C
Gauron
Carole
C
Kawakami
Koichi
K
Leucht
Christoph
C
Bally-Cuif
Laure
L
Volovitch
Michel
M
Bensimon
David
D
Jullien
Ludovic
L
Vriz
Sophie
S
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
D017931:DNA Primers; D018840:HSP70 Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; C045073:Cre recombinase; D019426:Integrases