Photoactivation of the CreER T2 recombinase for conditional site-specific recombination with high spatiotemporal resolution.

Publication Type:

Journal Article

Source:

Zebrafish, Volume 7, Issue 2, p.199-204 (2010)

DOI:

10.1089/zeb.2009.0632

Keywords:

Animals; Animals, Genetically Modified; DNA Primers; Gene Expression Regulation, Enzymologic; HSP70 Heat-Shock Proteins; Integrases; Microscopy, Fluorescence; Photochemical Processes; Polymerase Chain Reaction; Recombinant Fusion Proteins; Recombination, Genetic; Spectrometry, Fluorescence; Zebrafish

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).