Destabilized near-infrared fluorescent nanobodies enable background-free targeting of GFP-based biosensors for imaging and manipulation
Year: 2024
Type: article
Abstract: 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-FbLAG16 and NIR-FbLAG30, 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-FbLAGs bound to GCaMP6 enables ratiometric deep-brain in vivo imaging. Altogether, NIR-FbLAGs present a promising approach for imaging and manipulating various processes in live cells and behaving animals expressing GFP-based probes. Near-infrared (NIR) probes are in high demand as fluorescent tags for multicolour cellular and in vivo imaging. Here authors develop series of NIR nanobodies that bind to GFP-based biosensors, and demonstrate ratiometric deep-brain in vivo imaging of active GCaMP6f-expressing neurons.
Source: Nature Communications
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $6,790
Funders: U.S. Department of Health and Human Services, Jane ja Aatos Erkon Säätiö, Magnus Ehrnroothin Säätiö, National Research Center "Kurchatov Institute", National Institutes of Health, National Institute of General Medical Sciences, National Institute of Neurological Disorders and Stroke, Office of Extramural Research, National Institutes of Health
OpenAlex ID: W4402318511