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


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2007-11

AUTHORS

V Josserand, I Texier-Nogues, P Huber, M-C Favrot, J-L Coll

ABSTRACT

The bacterial lacZ gene encoding for beta-galactosidase (beta-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 beta-Gal expression in live cells in vitro or in vivo. The 9H-(1,3-dichloro-9,9-dimethylacridin- 2-one-7-yl) beta-D-galactopyranoside substrate was used to monitor beta-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 10(3) beta-Gal tumor cells located under the skin were detectable. In deeper organs like lung, as little as 5 ng of beta-Gal or Luciferase enzymes per mg of proteins were measured, confirming that both techniques reached similar sensibilities. Nonetheless, quantitative comparison of beta-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. More... »

PAGES

1587

References to SciGraph publications

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  • 2004-03. Targeting of lacZ reporter gene expression with radioiodine-labelled phenylethyl-β-d-thiogalactopyranoside in EUROPEAN JOURNAL OF NUCLEAR MEDICINE AND MOLECULAR IMAGING
  • 1999-09. Reporter gene vectors and assays in MOLECULAR BIOTECHNOLOGY
  • 2002-01. Scaling down imaging: molecular mapping of cancer in mice in NATURE REVIEWS CANCER
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  • 2005-08. Bioluminescent human breast cancer cell lines that permit rapid and sensitive in vivodetection of mammary tumors and multiple metastases in immune deficient mice in BREAST CANCER RESEARCH
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/sj.gt.3303028

    DOI

    http://dx.doi.org/10.1038/sj.gt.3303028

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1001619433

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/17882264


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