期刊
BIOTECHNOLOGY AND BIOENGINEERING
卷 112, 期 7, 页码 1483-1487出版社
WILEY
DOI: 10.1002/bit.25562
关键词
optogenetics; inducible expression; gene switch; TET system
资金
- European Community's Seventh Framework Programme [FP7/2007-2013]
- ERC [259043-CompBioMat]
- Excellence Initiative of the German Federal Government [EXC 294]
- Excellence Initiative of the German State Government [EXC 294]
The rapid development of mammalian optogenetics has produced an expanding number of gene switches that can be controlled with the unprecedented spatiotemporal resolution of light. However, in the pre-optogenetic era many networks, cell lines and transgenic organisms have been engineered that rely on chemically-inducible transgene expression systems but would benefit from the advantages of the traceless inducer light. To open the possibility for the effortless upgrade of such systems from chemical inducers to light, we capitalized on the specific Med25VBD inhibitor of the VP16/VP64 transactivation domain. In a first step, we demonstrated the efficiency and selectivity of Med25VBD in the inhibition of VP16/VP64-based transgene expression systems. Then, we fused the inhibitor to the blue light-responsive B-LID degron and optimized the performance of this construct with regard to the number of Med25VBD repeats. This approach resulted in an optogenetic upgrade of the popular Tet-OFF (TetR-VP64, tetO(7)-P-hCMVmin) system that allows tunable, blue light-inducible transgene expression in HEK-293T cells. Biotechnol. Bioeng. 2015;112: 1483-1487. (c) 2015 Wiley Periodicals, Inc.
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