Engineering of NEMO as calcium indicators with large dynamics and high sensitivity
Genetically encoded calcium indicators (GECIs) are indispensable tools for real-time monitoring of intracellular calcium signals and cellular activities in living organisms. Current GECIs face the challenge of suboptimal peak signal-to-baseline ratio (SBR) with limited resolution for reporting subtl...
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Veröffentlicht in: | Nature methods 2023-06, Vol.20 (6), p.918-924 |
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Sprache: | eng |
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Zusammenfassung: | Genetically encoded calcium indicators (GECIs) are indispensable tools for real-time monitoring of intracellular calcium signals and cellular activities in living organisms. Current GECIs face the challenge of suboptimal peak signal-to-baseline ratio (SBR) with limited resolution for reporting subtle calcium transients. We report herein the development of a suite of calcium sensors, designated NEMO, with fast kinetics and wide dynamic ranges (>100-fold). NEMO indicators report Ca
2+
transients with peak SBRs around 20-fold larger than the top-of-the-range GCaMP6 series. NEMO sensors further enable the quantification of absolution calcium concentration with ratiometric or photochromic imaging. Compared with GCaMP6s, NEMOs could detect single action potentials in neurons with a peak SBR two times higher and a median peak SBR four times larger in vivo, thereby outperforming most existing state-of-the-art GECIs. Given their high sensitivity and resolution to report intracellular Ca
2+
signals, NEMO sensors may find broad applications in monitoring neuronal activities and other Ca
2+
-modulated physiological processes in both mammals and plants.
The NEMO series of genetically encoded calcium indicators report calcium activity in neuronal and non-neuronal cells with high signal-to-baseline ratio, which is shown in neuronal culture, slice preparations, in vivo and in planta. |
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ISSN: | 1548-7091 1548-7105 |
DOI: | 10.1038/s41592-023-01852-9 |