SSBD:database

Project ID

476-Matsuda-GreenRedFRET

Project SID

476

Title

Calcium imaging of HeLa cells in response to histamine using improved Green-Red FRET sensors

Description

To monitor the calcium dynamics in cells, various genetically encoded calcium indicators (GECIs) based on FRET between fluorescent proteins are widely used for live imaging. Conventionally, cyan and yellow fluorescent proteins have been often used as FRET pairs. Meanwhile, bathochromically shifted indicators with green and red fluorescent protein pairs have various advantages, such as low toxicity and autofluorescence in cells. However, it remains difficult to develop them with a similar level of dynamic range as cyan and yellow fluorescent protein pairs. To improve this, we used Gamillus, which has a unique trans-configuration chromophore, as a green fluorescent protein. Based on one of the best high-dynamic-range GECIs, Twitch-NR, the authors developed a GECI with 1.5-times higher dynamic range, Twitch-GmRR, using RRvT as a red fluorescent protein. Twitch-GmRR had high brightness and photostability and was successfully applied for imaging the calcium dynamics in live cells. Our results suggest that Gamillus with trans-type chromophores contributes to improving the dynamic range of GECIs. Therefore, selection of the cis-trans isomer of the chromophore may be a fundamental approach to improve the dynamic range of green-red FRET indicators, unlimited by GECIs.

Submission Date

2025-09-30

Opened Date

-

Release Date

2026-06-09

Update Date

-

Kind

License

Project URL

-

Project DOI

-

Metadata Version

-

Template Version

3.0.1

Funding Information

This work was supported by MEXT Grant-in-Aid for Transformative Research Areas (A) “Integration of extracellular information by multimodal ECM activity” (No. 23H04935) to T.M., Scientific Research on Innovative Areas “Singularity biology” (No. 18H05410) to T.N., JST CREST Program (No. JPMJCR20E4) to T.M., and the grant of Joint Research by the National Institutes of Natural Sciences (NINS;No.01112105) to T.N.and K.O.K.O.was supported by Building of Consortia for the Development of Human Resources in Science and Technology, MEXT, Japan.

Total

6 files / 1.0 GB

Image datasets

zipped 2 files / 452.9 MB, raw image 4 files / 610.5 MB, total 6 files / 1.0 GB

Quantitative datasets

zipped 0 files / 0 bytes, raw bdml 0 files / 0 bytes, total 0 files / 0 bytes

Organism

Strain

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Cell

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Cell line

MeSH

Anatomical Entity

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UBERON

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Biological Process

Cellular Component

Molecular Function

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Medical Subject Headings

Method involved in biological imaging