Orthogonal Optogenetic Triple-Gene Control in Mammalian Cells
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- 作者：Konrad M眉ller ; Raphael Engesser ; Jens Timmer ; Matias D. Zurbriggen ; Wilfried Weber
- 刊名：ACS Synthetic Biology
- 出版年：2014
- 出版时间：November 21, 2014
- 年：2014
- 卷：3
- 期：11
- 页码：796-801
- 全文大小：292K
- ISSN：2161-5063
文摘
Optogenetic gene switches allow gene expression control at an unprecedented spatiotemporal resolution. Recently, light-responsive transgene expression systems that are activated by UV-B, blue, or red light have been developed. These systems perform well on their own, but their integration into genetic networks has been hampered by the overlapping absorbance spectra of the photoreceptors. We identified a lack of orthogonality between UV-B and blue light-controlled gene expression as the bottleneck and employed a model-based approach that identified the need for a blue light-responsive gene switch that is insensitive to low-intensity light. Based on this prediction, we developed a blue light-responsive and rapidly reversible expression system. Finally, we employed this expression system to demonstrate orthogonality between UV-B, blue, and red/far-red light-responsive gene switches in a single mammalian cell culture. We expect this approach to enable the spatiotemporal control of gene networks and to expand the applications of optogenetics in synthetic biology.