Bright and High-Performance Genetically Encoded Ca <sup>2+</sup> Indicator Based on mNeonGreen Fluorescent Protein
Landon Zarowny, Abhi Aggarwal, Virginia Rutten, Ilya Kolb, The GENIE Project, Ronak Patel, Hsin‐Yi Huang, Yu‐Fen Chang, Tiffany Phan, Richard Kanyo, Misha B. Ahrens, W. Ted Allison, Kaspar Podgorski, Robert E. Campbell
Abstract
Genetically encodable calcium ion (Ca2+) indicators (GECIs) based on green fluorescent proteins (GFP) are powerful tools for imaging of cell signaling and neural activity in model organisms. Following almost 2 decades of steady improvements in the Aequorea victoria GFP-based GCaMP series of GECIs, the performance of the most recent generation (i.e., jGCaMP7) may have reached its practical limit due to the inherent properties of GFP. In an effort to sustain the steady progression toward ever-improved GECIs, we undertook the development of a new GECI based on the bright monomeric GFP, mNeonGreen (mNG). The resulting indicator, mNG-GECO1, is 60% brighter than GCaMP6s in vitro and provides comparable performance as demonstrated by imaging Ca2+ dynamics in cultured cells, primary neurons, and in vivo in larval zebrafish. These results suggest that mNG-GECO1 is a promising next-generation GECI that could inherit the mantle of GCaMP and allow the steady improvement of GECIs to continue for generations to come.