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Engineering of NEMO as calcium indicators with large dynamics and high sensitivity
Jia Li1, Ziwei Shang2, Jia-Hui Chen3
1Beijing Key Laboratory of Gene Resource and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, China.
Nature Methods
|April 20, 2023
Summary
Researchers developed new genetically encoded calcium indicators (GECIs) called NEMO. These advanced sensors offer significantly improved signal-to-baseline ratios for precise monitoring of calcium signals in living cells.
Area of Science:
- Molecular Biology
- Neuroscience
- Biotechnology
Background:
- Genetically encoded calcium indicators (GECIs) are crucial for real-time monitoring of intracellular calcium signals.
- Existing GECIs often have limited signal-to-baseline ratio (SBR), hindering the detection of subtle calcium transients.
- Improved resolution is needed for accurate reporting of cellular activities.
Purpose of the Study:
- To develop novel GECIs with enhanced performance for calcium signal detection.
- To create sensors with fast kinetics, wide dynamic ranges, and superior SBR.
- To enable quantification of absolute calcium concentrations.
Main Methods:
- Development of a new suite of calcium sensors, designated NEMO.
- Characterization of NEMO indicators' kinetics, dynamic range, and SBR.
- In vivo comparison of NEMO sensors against GCaMP6s for detecting neuronal activity.
Main Results:
- NEMO indicators exhibit fast kinetics and >100-fold dynamic range.
- NEMO sensors achieve peak SBRs approximately 20-fold higher than GCaMP6 series.
- In vivo, NEMOs detected single action potentials with 2x higher peak SBR and 4x larger median peak SBR compared to GCaMP6s.
Conclusions:
- NEMO sensors significantly outperform existing state-of-the-art GECIs in sensitivity and resolution.
- NEMO's ability to quantify absolute calcium concentration offers advanced analytical capabilities.
- NEMO sensors are promising tools for diverse applications in neuroscience and plant biology.
