Temperature-responsive optogenetic probes of cell signaling.

Autor: Benman W; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA., Berlew EE; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA., Deng H; Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, PA, USA., Parker C; Department of Cell and Developmental Biology and Cardiovascular Institute, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA., Kuznetsov IA; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA., Lim B; Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, PA, USA.; Institute of Regenerative Medicine, University of Pennsylvania, Philadelphia, PA, USA., Siekmann AF; Department of Cell and Developmental Biology and Cardiovascular Institute, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, USA.; Institute of Regenerative Medicine, University of Pennsylvania, Philadelphia, PA, USA., Chow BY; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA., Bugaj LJ; Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA. bugaj@seas.upenn.edu.; Institute of Regenerative Medicine, University of Pennsylvania, Philadelphia, PA, USA. bugaj@seas.upenn.edu.; Abramson Cancer Center, University of Pennsylvania, Philadelphia, PA, USA. bugaj@seas.upenn.edu.
Jazyk: angličtina
Zdroj: Nature chemical biology [Nat Chem Biol] 2022 Feb; Vol. 18 (2), pp. 152-160. Date of Electronic Publication: 2021 Dec 22.
DOI: 10.1038/s41589-021-00917-0
Abstrakt: We describe single-component optogenetic probes whose activation dynamics depend on both light and temperature. We used the BcLOV4 photoreceptor to stimulate Ras and phosphatidyl inositol-3-kinase signaling in mammalian cells, allowing activation over a large dynamic range with low basal levels. Surprisingly, we found that BcLOV4 membrane translocation dynamics could be tuned by both light and temperature such that membrane localization spontaneously decayed at elevated temperatures despite constant illumination. Quantitative modeling predicted BcLOV4 activation dynamics across a range of light and temperature inputs and thus provides an experimental roadmap for BcLOV4-based probes. BcLOV4 drove strong and stable signal activation in both zebrafish and fly cells, and thermal inactivation provided a means to multiplex distinct blue-light sensitive tools in individual mammalian cells. BcLOV4 is thus a versatile photosensor with unique light and temperature sensitivity that enables straightforward generation of broadly applicable optogenetic tools.
(© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)
Databáze: MEDLINE
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