@article{cf33e7f8dd2c4be0b718d13662644700,
title = "Nuclear Localization Signals for Optimization of Genetically Encoded Tools in Neurons",
abstract = "Nuclear transport in neurons differs from that in non-neuronal cells. Here we developed a non-opsin optogenetic tool (OT) for the nuclear export of a protein of interest induced by near-infrared (NIR) light. In darkness, nuclear import reverses the OT action. We used this tool for comparative analysis of nuclear transport dynamics mediated by nuclear localization signals (NLSs) with different importin specificities. We found that widely used KPNA2-binding NLSs, such as Myc and SV40, are suboptimal in neurons. We identified uncommon NLSs mediating fast nuclear import and demonstrated that the performance of the OT for nuclear export can be adjusted by varying NLSs. Using these NLSs, we optimized the NIR OT for light-controlled gene expression for lower background and higher contrast in neurons. The selected NLSs binding importins abundant in neurons could improve performance of genetically encoded tools in these cells, including OTs and gene-editing tools.",
keywords = "importins, near-infrared, neurons, nuclear localization signal, nuclear transport, optogenetic tools",
author = "Karasev, {Maksim M.} and Mikhail Baloban and Verkhusha, {Vladislav V.} and Shcherbakova, {Daria M.}",
note = "Funding Information: We thank Olena Oliinyk (University of Helsinki) for useful suggestions. 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 grants GM122567 (to VV) and EY030705 (to DS) from the US National Institutes of Health, 322226 from the Academy of Finland (to VV), the Doctoral Programme in Biomedicine of the University of Helsinki (scholarship to MK), and by the grant from Orion Research Foundation (to MK). Funding Information: We thank Olena Oliinyk (University of Helsinki) for useful suggestions. 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 grants GM122567 (to VV) and EY030705 (to DS) from the US National Institutes of Health, 322226 from the Academy of Finland (to VV), the Doctoral Programme in Biomedicine of the University of Helsinki (scholarship to MK), and by the grant from Orion Research Foundation (to MK). Publisher Copyright: Copyright {\textcopyright} 2022 Karasev, Baloban, Verkhusha and Shcherbakova.",
year = "2022",
month = jul,
day = "19",
doi = "10.3389/fcell.2022.931237",
language = "English (US)",
volume = "10",
journal = "Frontiers in Cell and Developmental Biology",
issn = "2296-634X",
publisher = "Frontiers Media S. A.",
}