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

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)1121-1136
Number of pages16
JournalNature Protocols
Volume13
Issue number5
DOIs
StatePublished - May 1 2018

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Optogenetics
Protein Transport
Transcription
Multiplexing
Genes
Infrared radiation
Light
Proteins
Phytochrome
Cell membranes
Crosstalk
Reporter Genes
Gene expression
Biomedical Research
Cytoplasm
Cell Membrane
Oxygen
Gene Expression
Electric potential

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Near-infrared light-controlled systems for gene transcription regulation, protein targeting and spectral multiplexing. / Redchuk, Taras A.; Kaberniuk, Andrii; Verkhusha, Vladislav.

In: Nature Protocols, Vol. 13, No. 5, 01.05.2018, p. 1121-1136.

Research output: Contribution to journalArticle

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