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Metabolic biotinylation of cell surface receptors for in vivo imaging

Nature Methods volume 3, pages 391396 (2006) | Download Citation

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  • A Corrigendum to this article was published on 01 August 2006

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Abstract

We have developed a versatile, potent technique for imaging cells in culture and in vivo by expressing a metabolically biotinylated cell-surface receptor and visualizing it with labeled streptavidin moieties. The recombinant reporter protein, which incorporates a biotin acceptor peptide (BAP) between an N-terminal signal sequence and a transmembrane domain, (BAP-TM) was efficiently biotinylated by endogenous biotin ligase in mammalian cells with the biotin displayed on the cell surface. Tumors expressing the BAP-TM have high sensitivity for magnetic resonance and fluorescence tomographic imaging in vivo after intravascular injection of streptavidin conjugated to magnetic nanoparticles or fluorochromes, respectively. Moreover, streptavidin–horseradish peroxidase conjugates in conjunction with a peroxidase-sensitive gadolinium agent further increased and prolonged the magnetic resonance signal. This BAP-TM allows noninvasive real-time imaging of any cell type transduced to express this reporter protein in culture or in vivo.

*Note: In the version of this article originally published, reference 12 was incorrect. The correct reference 12 is Querol, M., Chen, J.W., Weissleder, R. & Bogdanov, A. Jr. DTPA-bis-amide based MR sensor agents for peroxidase imaging. Org. Lett. 17, 1719–1722 (2005). This error has been corrected in the PDF version of the article.

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Acknowledgements

This work was supported by grants from the US National Cancer Institute CA69246 (X.O.B. and R.W.), CA86355 (R.W. and X.O.B.), CA92782 (R.W.), U01 HL080731 (R.W.), R01 HL078641 (R.W.) and the Brain Tumor Society (X.O.B. and B.A.T.). We thank M. Sena-Esteves (MGH) for providing the lentivirus vectors, P. Waterman for FMT analysis, S. Rhee for magnetic resonance image acquisitions, M. Pittet for help with FACS analysis, N. Sergeyev for synthesizing MNP and F. Reynolds for synthesizing the Gd agent. We also thank S. McDavitt and M. Carlson for editorial assistance.

Author information

Author notes

    • Bakhos A Tannous
    •  & Jan Grimm

    These authors contributed equally to this work.

Affiliations

  1. Center for Molecular Imaging Research, Massachusetts General Hospital-East, Building 149, 13th Street, Charlestown, Massachusetts 02129, USA.

    • Bakhos A Tannous
    • , Jan Grimm
    • , John W Chen
    • , Ralph Weissleder
    •  & Xandra O Breakefield
  2. Department of Neurology, Massachusetts General Hospital-East, Building 149, 13th Street, Charlestown, Massachusetts 02129, USA.

    • Bakhos A Tannous
    • , Katherine F Perry
    •  & Xandra O Breakefield
  3. Neuroscience Program, Harvard Medical School, Boston, Massachusetts 02115, USA.

    • Bakhos A Tannous
    •  & Xandra O Breakefield

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Ralph Weissleder.

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DOI

https://doi.org/10.1038/nmeth875

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