Bright and High-Performance Genetically Encoded Ca2+ Indicator Based on mNeonGreen Fluorescent Protein
Landon Zarowny1, Abhi Aggarwal1,2, Virginia M S Rutten2,3
1Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
ACS Sensors
|June 24, 2020
Summary
Researchers developed mNG-GECO1, a new genetically encodable calcium indicator (GECI). This bright, GFP-based tool offers improved imaging of cell signaling and neural activity, potentially surpassing current GCaMP performance.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Genetically encodable calcium ion (Ca2+) indicators (GECIs) are crucial for observing cellular signaling and neural activity.
- The performance of existing green fluorescent protein (GFP)-based GCaMP indicators may be approaching inherent limitations.
Purpose of the Study:
- To develop a novel GECI with enhanced brightness and performance beyond the current GCaMP series.
- To create a successor to the GCaMP GECI line using a brighter fluorescent protein.
Main Methods:
- Engineered a new GECI, mNG-GECO1, utilizing the bright monomeric green fluorescent protein, mNeonGreen (mNG).
- Evaluated mNG-GECO1's performance in vitro, in cultured cells, primary neurons, and in vivo using larval zebrafish models.
Main Results:
- mNG-GECO1 demonstrated a 60% increase in brightness compared to GCaMP6s in vitro.
- Achieved comparable performance to GCaMP indicators in imaging Ca2+ dynamics across various biological systems.
- Successfully visualized Ca2+ signaling in cultured cells, primary neurons, and in vivo.
Conclusions:
- mNG-GECO1 represents a promising next-generation GECI with superior brightness.
- This new indicator has the potential to advance the field of calcium imaging, succeeding the GCaMP series.
- Continued development of GECIs is essential for future progress in understanding cellular processes.
