Destabilized near-infrared fluorescent nanobodies enable background-free targeting of GFP-based biosensors for imaging and manipulation.

Autor: Barykina NV; Department of Genetics, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, New York, NY, 10461, USA., Carey EM; Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA, 92037, USA., Oliinyk OS; Medicum, Faculty of Medicine, University of Helsinki, Helsinki, 00290, Finland., Nimmerjahn A; Waitt Advanced Biophotonics Center, Salk Institute for Biological Studies, La Jolla, CA, 92037, USA., Verkhusha VV; Department of Genetics, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, New York, NY, 10461, USA. vladislav.verkhusha@einsteinmed.edu.; Medicum, Faculty of Medicine, University of Helsinki, Helsinki, 00290, Finland. vladislav.verkhusha@einsteinmed.edu.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 Sep 06; Vol. 15 (1), pp. 7788. Date of Electronic Publication: 2024 Sep 06.
DOI: 10.1038/s41467-024-51857-x
Abstrakt: Near-infrared (NIR) probes are highly sought after as fluorescent tags for multicolor cellular and in vivo imaging. Here we develop small NIR fluorescent nanobodies, termed NIR-Fb LAG16 and NIR-Fb LAG30 , enabling background-free visualization of various GFP-derived probes and biosensors. We also design a red-shifted variant, NIR-Fb (718) , to simultaneously target several antigens within the NIR spectral range. Leveraging the antigen-stabilizing property of the developed NIR-Fbs, we then create two modular systems for precise control of gene expression in GFP-labeled cells. Applying the NIR-Fbs in vivo, we target cells expressing GFP and the calcium biosensor GCaMP6 in the somatosensory cortex of transgenic mice. Simultaneously tracking calcium activity and the reference signal from NIR-Fb LAG s bound to GCaMP6 enables ratiometric deep-brain in vivo imaging. Altogether, NIR-Fb LAG s present a promising approach for imaging and manipulating various processes in live cells and behaving animals expressing GFP-based probes.
(© 2024. The Author(s).)
Databáze: MEDLINE
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