@inbook{3e1d16198d004202a0dc6b52dcb47986,
title = "Light-controlled gene switches in mammalian cells",
abstract = "Remote control of cells is a desirable feature in synthetic biology. We established a light-switchable interfering peptide (iPEP) which controls gene expression by modulating the activity of a transcription factor. For photo-switching, the iPEP is cross-linked with a cis-trans isomerizable cross-linker in such a way that the light-activated cis form enables inhibitor folding rendering it active, whereas the dark-adapted trans form forces the inhibitor into an inactive form. Switching can be repeated in both directions. The iPEP acts as dominant-negative inhibitor targeting c-Jun and c-Fos of the transcription factor activator protein-1 (AP-1). Light-activated peptides exhibited much stronger inhibition of AP-1:DNA complexes and interference with gene transcription than their nonactivated counter parts. In this chapter, we provide protocols for cross-linking, peptide purification, observation of structural changes upon photo-switching, DNA binding analyses as well as gene expression studies in mammalian cells.",
keywords = "cis-trans Isomerization, Electrophoretic mobility shift assay, Inducible gene expression, Interfering peptide, Light-switchable",
author = "Fuzhong Zhang and M{\"u}ller, {Kristian M.} and Woolley, {G. Andrew} and Arndt, {Katja M.}",
year = "2012",
doi = "10.1007/978-1-61779-412-4_12",
language = "English",
isbn = "9781617794117",
series = "Methods in Molecular Biology",
pages = "195--210",
editor = "Wilfried Weber and Martin Fussenegger and Wilfried Weber",
booktitle = "Synthetic Gene Networks",
}