通过笼状环芬的光活化对蛋白质活性和基因表达进行光学控制。

Optical control of protein activity and gene expression by photoactivation of caged cyclofen.

作者信息

Hamouri Fatima, Zhang Weiting, Aujard Isabelle, Le Saux Thomas, Ducos Bertrand, Vriz Sophie, Jullien Ludovic, Bensimon David

机构信息

Laboratoire de Physique de l'ENS, CNRS-UMR8023, PSL Research University, Paris, France; Institut de Biologie de l'ENS, CNRS-UMR8197, INSERM-U1024, PSL Research University, Paris, France.

PASTEUR, Département de Chimie de l'ENS, CNRS, PSL Research University, Paris, France; Sorbonne Universités, UPMC Univ Paris 06, ENS, CNRS, PASTEUR, Paris, France.

出版信息

Methods Enzymol. 2019;624:1-23. doi: 10.1016/bs.mie.2019.04.009. Epub 2019 May 2.

DOI:10.1016/bs.mie.2019.04.009

PMID:31370925

Abstract

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.

摘要

利用光来控制基因表达和蛋白质活性是一个迅速发展的领域。虽然许多此类方法通过将光激活蛋白与目标蛋白融合来控制后者的活性，但也可以通过解除特定配体的笼蔽来控制蛋白质的活性。在这种情况下，通过解除其配体环芬醇（cyclofen-OH）的笼蔽来控制与修饰雌激素受体（ERT）融合的蛋白的激活，已成为一种在活生物体中定量、快速和局部控制蛋白质激活的通用且多功能的方法。在此，我们展示了这种方法背后的实验细节。