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

Author

Listed:
  • Natalia V. Barykina

    (Bronx)

  • Erin M. Carey

    (Salk Institute for Biological Studies)

  • Olena S. Oliinyk

    (University of Helsinki)

  • Axel Nimmerjahn

    (Salk Institute for Biological Studies)

  • Vladislav V. Verkhusha

    (Bronx
    University of Helsinki)

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.

Suggested Citation

  • Natalia V. Barykina & Erin M. Carey & Olena S. Oliinyk & Axel Nimmerjahn & Vladislav V. Verkhusha, 2024. "Destabilized near-infrared fluorescent nanobodies enable background-free targeting of GFP-based biosensors for imaging and manipulation," Nature Communications, Nature, vol. 15(1), pages 1-16, December.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-51857-x
    DOI: 10.1038/s41467-024-51857-x
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    References listed on IDEAS

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    2. Olena S. Oliinyk & Anton A. Shemetov & Sergei Pletnev & Daria M. Shcherbakova & Vladislav V. Verkhusha, 2019. "Smallest near-infrared fluorescent protein evolved from cyanobacteriochrome as versatile tag for spectral multiplexing," Nature Communications, Nature, vol. 10(1), pages 1-13, December.
    3. Jihoon Kim & Sangkyu Lee & Kanghoon Jung & Won Chan Oh & Nury Kim & Seungkyu Son & YoungJu Jo & Hyung-Bae Kwon & Won Do Heo, 2019. "Intensiometric biosensors visualize the activity of multiple small GTPases in vivo," Nature Communications, Nature, vol. 10(1), pages 1-11, December.
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