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ACS Synth Biol. 2015 Sep 18;4(9):951-8. doi: 10.1021/acssynbio.5b00004. Epub 2015 Mar 30.

Red Light-Regulated Reversible Nuclear Localization of Proteins in Mammalian Cells and Zebrafish.

Author information

1
Faculty of Biology, University of Freiburg , Schänzlestrasse 1, 79104 Freiburg, Germany.
2
BIOSS - Centre for Biological Signalling Studies, University of Freiburg , Schänzlestrasse 18, 79104 Freiburg, Germany.
3
SGBM - Spemann Graduate School of Biology and Medicine (SGBM), University of Freiburg , Albertstrasse 19a, 79104 Freiburg, Germany.
4
Institute of Toxicology and Genetics, Karlsruhe Institute of Technology and University of Heidelberg , D-76344 Eggenstein-Leopoldshafen, Germany.
5
BIF-IGS - BioInterfaces International Graduate School , Hermann-von-Helmholtz-Platz 1, 76344 Eggenstein-Leopoldshafen, Germany.
6
CCI, Centre for Chronic Immunodeficiency, University Clinincs Freiburg , Breisacher Strasse 117, 79106 Freiburg, Germany.
7
Biological Research Centre, Institute of Plant Biology , H-6726 Szeged, Hungary.
8
ZBSA - Centre for Biosystems Analysis, University of Freiburg , Habsburgerstrasse 49, 79104 Freiburg, Germany.

Abstract

Protein trafficking in and out of the nucleus represents a key step in controlling cell fate and function. Here we report the development of a red light-inducible and far-red light-reversible synthetic system for controlling nuclear localization of proteins in mammalian cells and zebrafish. First, we synthetically reconstructed and validated the red light-dependent Arabidopsis phytochrome B nuclear import mediated by phytochrome-interacting factor 3 in a nonplant environment and support current hypotheses on the import mechanism in planta. On the basis of this principle we next regulated nuclear import and activity of target proteins by the spatiotemporal projection of light patterns. A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression. These data demonstrate the first in vivo application of a plant phytochrome-based optogenetic tool in vertebrates and expand the repertoire of available light-regulated molecular devices.

KEYWORDS:

light-inducible nuclear transport; mammalian synthetic biology; optogenetics; phytochrome; plant synthetic biology

PMID:
25803699
DOI:
10.1021/acssynbio.5b00004
[Indexed for MEDLINE]

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