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MicroPET imaging of Cre–loxP-mediated conditional activation of a herpes simplex virus type 1 thymidine kinase reporter gene

Gene Therapy volume 11, pages 609–618 (2004)Cite this article

Abstract

Site-specific recombination tools such as the Cre–loxP system are used to create animal models where conditional gene deletion/activation studies are required. In the current proof of principle study, we have demonstrated that a PET reporter gene (PRG), the herpes simplex virus type 1 thymidine kinase (HSV1-tk), can be made to remain silent and can be activated by Cre–loxP-mediated recombination in cell culture and in living mice. An adenovirus carrying a silent HSV1-tk was tail-vein injected (1x109 PFU) in six transgenic mice that express Cre recombinase in their liver (Cre+) and in four control mice (Cre−). The liver-specific expression of the PRG in Cre+ mice was detected in the microPET following injection of the reporter probe, 9-[4-fluoro-3-(hydroxymethyl)butyl]guanine ([18F]-FHBG). The [18F]-FHBG accumulation in the liver in terms of percent-injected dose per gram of tissue was 7.72±1.13 for the Cre+ mice and 0.10±0.02 for the Cre− mice (P<0.05) 48 h after adenoviral injection. These results were further validated by quantitative RT-PCR, western blotting and by in vitro assays for herpes simplex virus type 1 thymidine kinase enzyme activity. Thus by using the Cre–loxP system it is possible to modulate a PRG and noninvasively monitor the extent of Cre–loxP-mediated gene activation by imaging in a microPET scanner.

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Acknowledgements

We thank Xiaoman Lewis, Judy Edwards and Waldmar Ladno (Crump Institute for Molecular Imaging) for technical assistance. We would also like to thank Dr Lily Wu and Jamie Matherly, UCLA for their help with adenoviral work. This work was supported by funding from Department of Energy contract DE-FC03-87ER60615 (SSG), NIH RO1 CA82214-01 (SSG) and SAIRP R24 CA92865 (SSG).

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Authors and Affiliations

  1. Crump Institute for Molecular Imaging, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA

    G Sundaresan, R Paulmurugan, F Berger & SS Gambhir

  2. Department of Molecular and Medical Pharmacology, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA

    G Sundaresan, R Paulmurugan, F Berger, B Stiles, H Wu & SS Gambhir

  3. Department of Pharmacology, Nagasaki University School of Medicine, Nagasaki, Japan

    Y Nagayama

  4. Jonsson Comprehensive Cancer Center, David Geffen School of Medicine at UCLA, Los Angeles, CA, USA

    SS Gambhir

  5. Department of Radiology and Bio-X Program, Molecular Imaging Program at Stanford, Stanford, CA, USA

    SS Gambhir

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  1. G Sundaresan
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Correspondence to SS Gambhir.

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Sundaresan, G., Paulmurugan, R., Berger, F. et al. MicroPET imaging of Cre–loxP-mediated conditional activation of a herpes simplex virus type 1 thymidine kinase reporter gene. Gene Ther 11, 609–618 (2004). https://doi.org/10.1038/sj.gt.3302194

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