We report an intensiometric, near-infrared fluorescent, genetically encoded calcium ion (Ca2+) indicator (GECI) with excitation and emission maxima at 678 and 704 nm, respectively. This GECI, designated NIR-GECO1, enables imaging of Ca2+ transients in cultured mammalian cells and brain tissue with sensitivity comparable to that of currently available visible-wavelength GECIs. We demonstrate that NIR-GECO1 opens up new vistas for multicolor Ca2+ imaging in combination with other optogenetic indicators and actuators.
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The NIR-GECO1 gene sequence is available through GenBank (submission no. MK134690). pDuEx2-NIR-GECO1 (plasmid no. 113,680) and pAAV-hSyn-NES-NIR-GECO1 (plasmid no. 113,683) are available via Addgene according to the terms of the Uniform Biological Material Transfer Agreement. Source data for Figs. 1–3 and Supplementary Figs. 5–14 are available online.
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The authors thank the University of Alberta Molecular Biology Services Unit; Y. Li, H. Zhou and A. Aggarwal, for technical support; A. Holt for providing access to the stopped-flow spectrophotometer; and M. Vanni, T. Murphy, A. Nimmerjahn and S. Chen for preliminary AAV testing. We thank M.-E. Paquet at the University of Laval Molecular Tools Platform and the Janelia Research Campus (JRC) Virus core for AAV production, We thank V. Rancic and the JRC Histology group for preparing cultured neurons. We thank D. Park and H.J. Suk for help with characterization of NIR-GECO1 in brain-slice and two-photon imaging. We thank M. Reiss for assistance with the mouse handling, X.L. Deán-Ben for help with the in vivo mesoscale data analysis, M. Davidson and X. Shu for the mIFP gene, and E. Rodriguez for the smURFP gene. Work in R.E.C.’s lab was supported by grants from NSERC (RGPIN 288338-2010), CIHR (MOP 123514 and FS 154310), Brain Canada and NIH (UO1 NS090565). D.R. acknowledges support from the European Research Council (ERC-2015-CoG-682379). The work of D.R. and S.S was also supported by the NIH (R21-EY026382 and UF1-NS107680). Work in J.Z.’s lab was supported by NIH (R01-DK073368 and R35-CA197622). E.S.B. was supported by J. Doerr, the HHMI-Simons Faculty Scholars Program, the Open Philanthropy Project, Human Frontier Science Program (RGP0015/2016), US Army Research Laboratory and the US Army Research Office (W911NF1510548), US-Israel Binational Science Foundation (2014509) and NIH (2R01-DA029639 and 1R01-GM104948).