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Nucleic Acids Res. 2017 Nov 16;45(20):e172. doi: 10.1093/nar/gkx804.

Optimized light-inducible transcription in mammalian cells using Flavin Kelch-repeat F-box1/GIGANTEA and CRY2/CIB1.

Quejada JR1,2,3, Park SE1,2, Awari DW1,2, Shi F1,2,4, Yamamoto HE1,2,5, Kawano F1,2, Jung JC6, Yazawa M1,2,3.

Author information

1
Columbia Stem Cell Initiative, Columbia University, New York, NY 10032, USA.
2
Department of Rehabilitation and Regenerative Medicine, Columbia University, New York, NY 10032, USA.
3
Department of Pharmacology, Columbia University, New York, NY 10032, USA.
4
College of Precision Instrument and Optoelectronics Engineering, Tianjin University, Tianjin 300072, China.
5
Barnard College, New York, NY 10027, USA.
6
Department of Biology, Stanford University, Stanford, CA 94305, USA.

Abstract

Light-inducible systems allow spatiotemporal control of a variety of biological activities. Here, we report newly optimized optogenetic tools to induce transcription with light in mammalian cells, using the Arabidopsis photoreceptor Flavin Kelch-repeat F-box 1 (FKF1) and its binding partner GIGANTEA (GI) as well as CRY2/CIB1. By combining the mutagenesis of FKF1 with the optimization of a split FKF1/GI dimerized Gal4-VP16 transcriptional system, we identified constructs enabling significantly improved light-triggered transcriptional induction. In addition, we have improved the CRY2/CIB1-based light-inducible transcription with split construct optimization. The improvements regarding the FKF1/GI- and CRY2/CIB1-based systems will be widely applicable for the light-dependent control of transcription in mammalian cells.

PMID:
29040770
PMCID:
PMC5714181
DOI:
10.1093/nar/gkx804
[Indexed for MEDLINE]
Free PMC Article

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