Journal
ACS SYNTHETIC BIOLOGY
Volume 7, Issue 5, Pages 1349-1358Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.7b00450
Keywords
optogenetics; light-responsive gene expression; green light; CarH; AdoB12; plants
Categories
Funding
- Excellence Initiative of the German Federal Government [BIOSS EXC-294, CEPLAS EXC-1028, SGBM GSC-4, iGRAD-Plant GRK-1525]
- Excellence Initiative of the German State Government [BIOSS EXC-294, CEPLAS EXC-1028, SGBM GSC-4, iGRAD-Plant GRK-1525]
- DFG [WE 4733/4-1, SPP1926 Z259/2-1]
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The ever-increasing complexity of synthetic gene networks and applications of synthetic biology requires precise and orthogonal gene expression systems. Of particular interest are systems responsive to light as they enable the control of gene expression dynamics with unprecedented resolution in space and time. While broadly used in mammalian backgrounds, however, optogenetic approaches in plant cells are still limited due to interference of the activating light with endogenous photoreceptors. Here, we describe the development of the first synthetic light-responsive system for the targeted control of gene expression in mammalian and plant cells that responds to the green range of the light spectrum in which plant photoreceptors have minimal activity. We first engineered a system based on the light-sensitive bacterial transcription factor CarH and its cognate DNA operator sequence CarO from Thermus thermophilus to control gene expression in mammalian cells. The system was functional in various mammalian cell lines, showing high induction (up to 350-fold) along with low leakiness, as well as high reversibility. We quantitatively described the systems characteristics by the development and experimental validation of a mathematical model. Finally, we transferred the system into A. thaliana protoplasts and demonstrated gene repression in response to green light. We expect that this system will provide new opportunities in applications based on synthetic gene networks and will open up perspectives for optogenetic studies in mammalian and plant cells.
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