Near-Infrared Light-Controlled Gene Expression and Protein Targeting in Neurons and Non-neuronal Cells

Chembiochem : a European Journal of Chemical Biology
Taras A RedchukVladislav V Verkhusha

Abstract

Near-infrared (NIR) light-inducible binding of bacterial phytochrome BphP1 to its engineered partner, QPAS1, is used for optical protein regulation in mammalian cells. However, there are no data on the application of the BphP1-QPAS1 pair in cells derived from various mammalian tissues. Here, we tested the functionality of two BphP1-QPAS1-based optogenetic tools-an NIR- and blue-light-sensing system for control of protein localization (iRIS) and an NIR light-sensing system for transcription activation (TA)-in several cell types, including cortical neurons. We found that the performance of these optogenetic tools often relied on physiological properties of a specific cell type, such as nuclear transport, which could limit the applicability of the blue-light-sensitive component of iRIS. In contrast, the NIR-light-sensing component of iRIS performed well in all tested cell types. The TA system showed the best performance in cervical cancer (HeLa), bone cancer (U-2 OS), and human embryonic kidney (HEK-293) cells. The small size of the QPAS1 component allowed the design of adeno-associated virus (AAV) particles, which were applied to deliver the TA system to neurons.

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Citations

Jul 25, 2018·Critical Reviews in Biochemistry and Molecular Biology·Robert M Hughes
Jul 2, 2019·Human Molecular Genetics·Claire Domenger, Dirk Grimm
Mar 10, 2019·Nature Communications·Anna V LeopoldVladislav V Verkhusha
Feb 1, 2020·Nature Communications·Taras A RedchukVladislav V Verkhusha
Nov 24, 2018·Frontiers in Microbiology·Zedao LiuQuanfeng Liang
Oct 13, 2020·Advanced Biology·Markus M KramerGerald Radziwill
Aug 28, 2021·Nature Methods·Kyrylo Yu ManoilovDaria M Shcherbakova

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