TY - JOUR
T1 - Near-Infrared Light-Controlled Gene Expression and Protein Targeting in Neurons and Non-neuronal Cells
AU - Redchuk, Taras A.
AU - Karasev, Maksim M.
AU - Omelina, Evgeniya S.
AU - Verkhusha, Vladislav V.
N1 - Funding Information:
We thank the Biomedicum Imaging Unit, the Neuronal Cell Culture Unit, and the AAV Gene Transfer and Cell Therapy core facilities of the University of Helsinki for technical assistance. This work was supported by the National Institutes of Health (NIH, grants R35 GM 122567 and U01 NS 103573), the EU FP7 Program (ERC-2013-ADG-340233), and the Academy of Finland (263371 and 266992).
Funding Information:
We thank the Biomedicum Imaging Unit, the Neuronal Cell Culture Unit, and the AAV Gene Transfer and Cell Therapy core facilities of the University of Helsinki for technical assistance. This work was supported by the National Institutes of Health (NIH, grants R35 GM 122567 and U01 NS103573), the EU FP7 Program (ERC-2013-ADG-340233), and the Academy of Finland (263371 and 266992).
Publisher Copyright:
© 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
PY - 2018/6/18
Y1 - 2018/6/18
N2 - 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.
AB - 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.
KW - BphP1
KW - QPAS1
KW - gene expression
KW - near-infrared
KW - optogenetic
UR - http://www.scopus.com/inward/record.url?scp=85047337236&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85047337236&partnerID=8YFLogxK
U2 - 10.1002/cbic.201700642
DO - 10.1002/cbic.201700642
M3 - Article
C2 - 29465801
AN - SCOPUS:85047337236
VL - 19
SP - 1334
EP - 1340
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 12
ER -