A highly-sensitive genetically encoded temperature indicator exploiting a temperature-responsive elastin-like polypeptide

Genetically encoded temperature indicators (GETIs) allow for real-time measurement of subcellular temperature dynamics in live cells. However, GETIs have suffered from poor temperature sensitivity, which may not be sufficient to resolve small heat production from a biological process. Here, we devel...

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Bibliographic Details
Published in:Scientific reports 2021-08, Vol.11 (1), p.16519-16519, Article 16519
Main Authors: Vu, Cong Quang, Fukushima, Shun-ichi, Wazawa, Tetsuichi, Nagai, Takeharu
Format: Article
Language:English
Subjects:
631/1647
631/57
631/80
631/92
Calcium influx
Cytoplasm
Elastin
Energy transfer
Fluorescence resonance energy transfer
Heat
High temperature
Humanities and Social Sciences
Macromolecules
multidisciplinary
Phase transitions
Polypeptides
Science
Science (multidisciplinary)
Temperature
Temperature measurement
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Summary:Genetically encoded temperature indicators (GETIs) allow for real-time measurement of subcellular temperature dynamics in live cells. However, GETIs have suffered from poor temperature sensitivity, which may not be sufficient to resolve small heat production from a biological process. Here, we develop a highly-sensitive GETI, denoted as ELP-TEMP, comprised of a temperature-responsive elastin-like polypeptide (ELP) fused with a cyan fluorescent protein (FP), mTurquoise2 (mT), and a yellow FP, mVenus (mV), as the donor and acceptor, respectively, of Förster resonance energy transfer (FRET). At elevated temperatures, the ELP moiety in ELP-TEMP undergoes a phase transition leading to an increase in the FRET efficiency. In HeLa cells, ELP-TEMP responded to the temperature from 33 to 40 °C with a maximum temperature sensitivity of 45.1 ± 8.1%/°C, which was the highest ever temperature sensitivity among hitherto-developed fluorescent nanothermometers. Although ELP-TEMP showed sensitivity not only to temperature but also to macromolecular crowding and self-concentration, we were able to correct the output of ELP-TEMP to achieve accurate temperature measurements at a subcellular resolution. We successfully applied ELP-TEMP to accurately measure temperature changes in cells induced by a local heat spot, even if the temperature difference was as small as 
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-021-96049-5