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Non-invasive in vivo optical imaging of the lacZ and luc gene expression in mice

Gene Therapy volume 14, pages 1587–1593 (2007)Cite this article

Abstract

The bacterial lacZ gene encoding for β-galactosidase (β-gal) is a common reporter gene used in transgenic mice. Nonetheless, the absence of fluorigenic substrates usable in live animals greatly hampered the non-invasive follow-up of this reporter gene expression. We used far-red fluorescence for imaging β-Gal expression in live cells in vitro or in vivo. The 9H-(1,3-dichloro-9,9-dimethylacridin- 2-one-7-yl) β-D-galactopyranoside substrate was used to monitor β-Gal expression as a reporter of tumor growth, or of the physiological levels of an endogenous gene or of gene transfer in lung. A quantitative evaluation of this method as well as a comparison of its sensitivity with Firefly Luciferase-based bioluminescence was also performed. In vivo measurements showed that 103 β-Gal tumor cells located under the skin were detectable. In deeper organs like lung, as little as 5 ng of β-Gal or Luciferase enzymes per mg of proteins were measured, confirming that both techniques reached similar sensibilities. Nonetheless, quantitative comparison of β-Gal levels measured with far-red imaging or with a standardized enzymatic evaluation after killing revealed that the 2D-fluorescent reflectance imaging method is submitted to a color-dependent disparity of the organs and cannot supply quantitative measurements but that a simple correction can be applied.

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Acknowledgements

This work was supported by the Institut National de la Sante Et de la Recherche Medicale (INSERM), the programme Interdisciplinaire 2001–2004 ‘Imagerie du Petit Animal’, the Canceropole Lyon-Alpes Rhone-alpes (CLARA), the Association for Research on Cancer (ARC), the Agence Nationale pour la Recherche (ANR) and the EMIL and N2L NoEs of the 6th framework. We are also grateful to Philippe Rizo (CEA Leti, France) for offering optical imaging system and to Dr Anne Imberty for useful discussion. We thank Corine Tenaud, Mélanie Guidetti and Dominique Desplanques in our laboratory for technical assistance.

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Authors and Affiliations

  1. INSERM U823, Institut Albert Bonniot, La Tronche Cedex, France

    V Josserand, M-C Favrot & J-L Coll

  2. Université Joseph Fourier, Grenoble Cedex, France

    V Josserand, P Huber, M-C Favrot & J-L Coll

  3. ANIMAGE, CERMEP, 59 boulevard Pinel, Lyon, France

    V Josserand

  4. LETI-DTBS, CEA-Grenoble, 17 rue des Martyrs, Grenoble Cedex, France

    I Texier-Nogues

  5. INSERM U882, CEA-Grenoble, Grenoble, France

    P Huber

  6. CEA iRTSV-APV, CEA-Grenoble, Grenoble, France

    P Huber

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  1. V Josserand
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  2. I Texier-Nogues
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  3. P Huber
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  5. J-L Coll
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Correspondence to J-L Coll.

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Josserand, V., Texier-Nogues, I., Huber, P. et al. Non-invasive in vivo optical imaging of the lacZ and luc gene expression in mice. Gene Ther 14, 1587–1593 (2007). https://doi.org/10.1038/sj.gt.3303028

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  • DOI: https://doi.org/10.1038/sj.gt.3303028

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