110024-82-3Relevant articles and documents
Optical control of protein activity and gene expression by photoactivation of caged cyclofen
Hamouri, Fatima,Zhang, Weiting,Aujard, Isabelle,Le Saux, Thomas,Ducos, Bertrand,Vriz, Sophie,Jullien, Ludovic,Bensimon
, p. 1 - 23 (2019)
The use of light to control the expression of genes and the activity of proteins is a rapidly expanding field. While many of these approaches use a fusion between a light activatable protein and the protein of interest to control the activity of the latter, it is also possible to control the activity of a protein by uncaging a specific ligand. In that context, controlling the activation of a protein fused to the modified estrogen receptor (ERT) by uncaging its ligand cyclofen-OH has emerged as a generic and versatile method to control the activation of proteins quantitatively, quickly and locally in a live organism. Here, we present the experimental details behind this approach.
Photocontrol of protein activity in cultured cells and zebrafish with one- and two-photon illumination
Sinha, Deepak Kumar,Neveu, Pierre,Gagey, Nathalie,Aujard, Isabelle,Benbrahim-Bouzidi, Chouaha,Le Saux, Thomas,Rampon, Christine,Gauron, Carole,Goetz, Bernard,Dubruille, Sylvie,Baaden, Marc,Volovitch, Michel,Bensimon, David,Vriz, Sophie,Jullien, Ludovic
, p. 653 - 663 (2011/03/17)
We have implemented a noninvasive optical method for the fast control of protein activity in a live zebrafish embryo. It relies on releasing a protein fused to a modified estrogen receptor ligand binding domain from its complex with cytoplasmic chaperones, upon the local photoactivation of a nonendogenous caged inducer. Molecular dynamics simulations were used to design cyclofen-OH, a photochemically stable inducer of the receptor specific for 4-hydroxy-tamoxifen (ERT2). Cyclofen-OH was easily synthesized in two steps with good yields. At submicromolar concentrations, it activates proteins fused to the ERT2 receptor. This was shown in cultured cells and in zebrafish embryos through emission properties and subcellular localization of properly engineered fluorescent proteins. Cyclofen-OH was successfully caged with various photolabile protecting groups. One particular caged compound was efficient in photoinducing the nuclear translocation of fluorescent proteins either globally (with 365 nm UV illumination) or locally (with a focused UV laser or with two-photon illumination at 750 nm). The present method for photocontrol of protein activity could be used more generally to investigate important physiological processes (e.g., in embryogenesis, organ regeneration and carcinogenesis) with high spatiotemporal resolution.