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A reporter system for replication-competent gammaretroviruses: the inGluc-MLV-DERSE assay

Gene Therapy volume 20, pages 169–176 (2013)Cite this article

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Abstract

Although novel retroviral vectors for use in gene-therapy products are reducing the potential for formation of replication-competent retrovirus (RCR), it remains crucial to screen products for RCR for both research and clinical purposes. For clinical-grade gammaretrovirus-based vectors, RCR screening is achieved by an extended S+L− or marker-rescue assay, whereas standard methods for replication-competent lentivirus detection are still in development. In this report, we describe a rapid and sensitive method for replication-competent gammaretrovirus detection. We used this assay to detect three members of the gammaretrovirus family and compared the sensitivity of our assay with well-established methods for retrovirus detection, including the extended S+L− assay. Results presented here demonstrate that this assay should be useful for gene-therapy product testing.

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Acknowledgements

We thank Jane Mirro, Gerry Princler, Patricia Lloyd and Shawn Hill for superb technical assistance, and Vineet KewalRamani for many helpful discussions. This work was supported in part by the Intramural Research Program of the NIH, National Cancer Institute and Center for Cancer Research. Work at Indiana University is supported by the NIH, National Center for Research Resource (National Gene Vector Biorepository P40 RR024928).

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Author notes

  1. A L Aloia

    Present address: 5Current address: School of Molecular and Biomedical Science, The University of Adelaide, Adelaide, South Australia, Australia.,

  2. V Pak

    Present address: 6Current address: Department of Medicine, University of California at San Francisco, San Francisco, CA, USA.,

  3. D Derse: Deceased.

Authors and Affiliations

  1. HIV Drug Resistance Program, National Cancer Institute, Frederick, MD, USA

    A L Aloia, V Pak, K E Lee, S Sanchez-Martinez, D Derse, G Heidecker & A Rein

  2. Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN, USA

    L Duffy & K Cornetta

  3. Department of Medicine, Indiana University School of Medicine, Indianapolis, IN, USA

    K Cornetta

  4. Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN, USA

    K Cornetta

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  1. A L Aloia
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  2. L Duffy
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  7. G Heidecker
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  8. K Cornetta
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  9. A Rein
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Correspondence to A Rein.

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Aloia, A., Duffy, L., Pak, V. et al. A reporter system for replication-competent gammaretroviruses: the inGluc-MLV-DERSE assay. Gene Ther 20, 169–176 (2013). https://doi.org/10.1038/gt.2012.18

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