Chapter title |
Light-controlled gene switches in Mammalian cells.
|
---|---|
Chapter number | 12 |
Book title |
Synthetic Gene Networks
|
Published in |
Methods in molecular biology, January 2012
|
DOI | 10.1007/978-1-61779-412-4_12 |
Pubmed ID | |
Book ISBNs |
978-1-61779-411-7, 978-1-61779-412-4
|
Authors |
Fuzhong Zhang, Kristian M. Müller, G. Andrew Woolley, Katja M. Arndt, Zhang, Fuzhong, Müller, Kristian M., Woolley, G. Andrew, Arndt, Katja M. |
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. |
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Members of the public | 2 | 100% |
Mendeley readers
Geographical breakdown
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United States | 1 | 7% |
Unknown | 13 | 93% |
Demographic breakdown
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Student > Ph. D. Student | 4 | 29% |
Professor | 3 | 21% |
Researcher | 3 | 21% |
Student > Master | 2 | 14% |
Other | 1 | 7% |
Other | 0 | 0% |
Unknown | 1 | 7% |
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Biochemistry, Genetics and Molecular Biology | 4 | 29% |
Chemistry | 1 | 7% |
Medicine and Dentistry | 1 | 7% |
Unknown | 1 | 7% |