Near-infrared light-controlled systems for gene transcription regulation, protein targeting and spectral multiplexing

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Redchuk , T A , Kaberniuk , A A & Verkhusha , V V 2018 , ' Near-infrared light-controlled systems for gene transcription regulation, protein targeting and spectral multiplexing ' , Nature Protocols , vol. 13 , no. 5 , pp. 1121-1136 . https://doi.org/10.1038/nprot.2018.022

Title: Near-infrared light-controlled systems for gene transcription regulation, protein targeting and spectral multiplexing
Author: Redchuk, Taras A.; Kaberniuk, Andrii A.; Verkhusha, Vladislav V.
Contributor: University of Helsinki, Medicum
University of Helsinki, Medicum
Date: 2018-05
Language: eng
Number of pages: 16
Belongs to series: Nature Protocols
ISSN: 1754-2189
URI: http://hdl.handle.net/10138/301886
Abstract: Near-infrared (NIR, 740-780 nm) optogenetic systems are well-suited to spectral multiplexing with blue-light-controlled tools. Here, we present two protocols, one for regulation of gene transcription and another for control of protein localization, that use a NIR-responsive bacterial phytochrome BphP1-QPAS1 optogenetic pair. In the first protocol, cells are transfected with the optogenetic constructs for independently controlling gene transcription by NIR (BphP1-QPAS1) and blue (LightOn) light. The NIR and blue-light-controlled gene transcription systems show minimal spectral crosstalk and induce a 35- to 40-fold increase in reporter gene expression. In the second protocol, the BphP1-QPAS1 pair is combined with a light-oxygen-voltage-sensing (LOV) domain-based construct into a single optogenetic tool, termed iRIS. This dual-light-controllable protein localization tool allows tridirectional protein translocation among the cytoplasm, nucleus and plasma membrane. Both procedures can be performed within 3-5 d. Use of NIR light-controlled optogenetic systems should advance basic and biomedical research.
Subject: SPATIOTEMPORAL CONTROL
IN-VIVO
FLUORESCENT PROTEINS
IMAGE-ANALYSIS
RED-LIGHT
BACTERIAL PHYTOCHROMES
INDUCIBLE EXPRESSION
OPTOGENETIC CONTROL
TRANSGENIC MICE
ZEBRAFISH
1182 Biochemistry, cell and molecular biology
3111 Biomedicine
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