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A Method to Rapidly and Accurately Compare the Relative Efficacies of Non-invasive Imaging Reporter Genes in a Mouse Model and its Application to Luciferase Reporters

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Abstract

Purpose

Our goal is to develop a simple, quantitative, robust method to compare the efficacy of imaging reporter genes in culture and in vivo. We describe an adenoviral vector–liver transduction procedure and compare the luciferase reporter efficacies.

Procedures

Alternative reporter genes are expressed in a common adenoviral vector. Vector amounts used in vivo are based on cell culture titrations, ensuring that the same transduction efficacy is used for each vector. After imaging, in vivo and in vitro values are normalized to hepatic vector transduction using quantitative real-time PCR.

Results

We assayed standard firefly luciferase (FLuc), enhanced firefly luciferase (EFLuc), luciferase 2 (Luc2), humanized Renilla luciferase (hRLuc), Renilla luciferase 8.6-535 (RLuc8.6), and a membrane-bound Gaussia luciferase variant (extGLuc) in cell culture and in vivo. We observed greater than 100-fold increase in bioluminescent signal for both EFLuc and Luc2 when compared to FLuc and greater than 106-fold increase for RLuc8.6 when compared to hRLuc. ExtGLuc was not detectable in liver.

Conclusions

Our findings contrast, in some cases, with conclusions drawn in prior comparisons of these reporter genes and demonstrate the need for a standardized method to evaluate alternative reporter genes in vivo. Our procedure can be adapted for reporter genes that utilize alternative imaging modalities (fluorescence, bioluminescence, MRI, SPECT, PET).

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Acknowledgements

We thank Arthur Catapang for technical help, David Stout and Waldemar Ladno for advice and assistance with the optical imaging experiments, and Arion Chattziioannou and the members of the Herschman lab for helpful discussions. This study was funded by the National Cancer Institute In Vivo Cellular and Molecular Imaging Center (ICMIC) award P50 CA086306 (HRH). JG is supported by a Scholars in Oncologic Medical Imaging (SOMI) fellowship from the National Cancer Institute (Award R25T CA098010).

Conflicts of Interest

The authors declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Molecular Biology Institute, David Geffen School of Medicine, UCLA, 341 Boyer Hall, 611 Charles E Young Drive, Los Angeles, CA, 90095-1570, USA

    Jose S. Gil, Hidevaldo B. Machado & Harvey R. Herschman

  2. Department of Biological Chemistry, UCLA, 341 Boyer Hall, 611 Charles E Young Drive, Los Angeles, CA, 90095-1570, USA

    Jose S. Gil, Hidevaldo B. Machado & Harvey R. Herschman

  3. Department of Pharmacology, UCLA, 341 Boyer Hall, 611 Charles E Young Drive, Los Angeles, CA, 90095-1570, USA

    Jose S. Gil, Hidevaldo B. Machado & Harvey R. Herschman

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  1. Jose S. Gil
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  2. Hidevaldo B. Machado
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  3. Harvey R. Herschman
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Correspondence to Harvey R. Herschman.

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Gil, J.S., Machado, H.B. & Herschman, H.R. A Method to Rapidly and Accurately Compare the Relative Efficacies of Non-invasive Imaging Reporter Genes in a Mouse Model and its Application to Luciferase Reporters. Mol Imaging Biol 14, 462–471 (2012). https://doi.org/10.1007/s11307-011-0515-1

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