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Monitoring adenoviral DNA delivery, using a mutant herpes simplex virus type 1 thymidine kinase gene as a PET reporter gene

Gene Therapy volume 9, pages 1659–1666 (2002)Cite this article

Abstract

Current gene therapy protocols often suffer from an inability to monitor the site, level and persistence of gene expression following somatic DNA delivery. Herpes simplex virus 1 thymidine kinase (HSV1-tk) is currently under intensive investigation as a reporter gene for in vivo imaging of reporter gene expression. The presence of the HSV1-tk reporter gene is repetitively and non-invasively monitored by systemic injection of positron-emitting, radionuclide-labeled thymidine analogues or acycloguanosine HSV1-TK substrates and subsequent detection, by positron emission tomography, of trapped, phosphorylated product. To improve the efficacy of the HSV1-tk PET reporter gene system, both alternative substrates and mutations in the HSV1-tk gene have been described. We used a replication defective adenovirus to deliver the HSV1-sr39tk mutant enzyme and the wild-type HSV1-tk enzyme to mice. HSV1-sr39TK demonstrates greater sensitivity than wild-type HSV1-TK enzyme in vivo, using 9-[(4-[18F]fluoro-3-hydroxymethylbutyl)guanine as probe, following adenovirus-mediated hepatic expression in mice. Using this adenoviral delivery system, the location, magnitude and duration of HSV1-sr39tk PET reporter gene expression could be non-invasively, quantitatively and repetitively monitored for over 3 months by microPET.

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Acknowledgements

We thank Art Catapang, Raymond Basconcillo, Judy Edwards, Waldemar Ladno and ManKit Ho for technical assistance and Richard Leahy for consultation. This work was supported by Department of Energy award DE-FC03-87ER60615 (HRH, MEP, SSG, JRB, NS), NIH award P50 CA86306 (HRH, MEP, SSG, JRB, NS), NIH award R0-1 CA84572 (HRH) and NIH award R0-1 CA82214-01 (SSG).

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Authors and Affiliations

  1. Crump Institute for Molecular Imaging, UCLA School of Medicine, Los Angles, CA, USA

    Q Liang, K Nguyen, M E Phelps, S S Gambhir & H R Herschman

  2. UCLA/DOE Laboratory of Structural Biology and Molecular Medicine, UCLA School of Medicine, Los Angles, CA, USA

    N Satyamurthy, J R Barrio, M E Phelps, S S Gambhir & H R Herschman

  3. Department of Molecular and Medical Pharmacology, UCLA School of Medicine, Los Angles, CA, USA

    Q Liang, K Nguyen, N Satyamurthy, J R Barrio, M E Phelps, S S Gambhir & H R Herschman

  4. Molecular Biology Institute, UCLA School of Medicine, Los Angles, CA, USA

    H R Herschman

  5. Department of Biomathematics, UCLA School of Medicine, Los Angles, CA, USA

    M E Phelps & S S Gambhir

  6. UCLA-Jonsson Comprehensive Cancer Center, UCLA School of Medicine, Los Angles, CA, USA

    M E Phelps, S S Gambhir & H R Herschman

  7. Department of Biological Chemistry, UCLA School of Medicine, Los Angeles, CA, USA

    H R Herschman

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  1. Q Liang
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Liang, Q., Nguyen, K., Satyamurthy, N. et al. Monitoring adenoviral DNA delivery, using a mutant herpes simplex virus type 1 thymidine kinase gene as a PET reporter gene. Gene Ther 9, 1659–1666 (2002). https://doi.org/10.1038/sj.gt.3301899

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  • DOI: https://doi.org/10.1038/sj.gt.3301899

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