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A method to codetect introduced genes and their products in gene therapy protocols

Nature Biotechnology volume 14, pages 1012–1016 (1996)Cite this article

Abstract

To monitor the presence of introduced genes and the distribution of the encoded proteins in host tissues after gene transfer, we combined fluorescence in situ hybridization (FISH) and immunohistochemistry in two separate gene therapy paradigms. In brain tissue sections from animals injected with pHSVIac vector, we localized nuclei containing vector DMA both in cells expressing and not expressing ß-galactosidase (ß-gal). This suggests that the efficiency of gene transfer is affected not only by gene delivery, but also by cellular controls on gene expression. In a second paradigm, following myoblast transplantation, we detected donor nuclei in the muscle of a patient with Duchenne's muscular dystrophy. The donor nuclei were either surrounded by host nuclei or apparently fused in the patient's muscle fiber producing dystrophin. The combined FISH and immunohistochemistry assay offers greater sensitivity and more information than currently used polymerase chain reaction and protein detection methods.

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Author notes

  1. Louis M. Kunkel: kunkel@rascal.med.harvard.edu

Authors and Affiliations

  1. Division of Genetics and the Howard Hughes Medical Institute at Children's Hospital, 300 Longwood Ave., Boston, MA, 02115

    Emanuela Gussoni & Louis M. Kunkel

  2. Division of Endocrinology at Children's Hospital, 300 Longwood Ave., Boston, MA, 02115

    Yaming Wang, Cornel Fraefel & Alfred I. Geller

  3. Department of Neurology, California Pacific Medical Center, San Francisco, CA, 94115

    Robert G. Miller

  4. Department of Molecular Pharmacology, Stanford University School of Medicine, Stanford, CA, 94305

    Helen M. Blau

Authors
  1. Emanuela Gussoni
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  2. Yaming Wang
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  3. Cornel Fraefel
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  6. Alfred I. Geller
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  7. Louis M. Kunkel
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Gussoni, E., Wang, Y., Fraefel, C. et al. A method to codetect introduced genes and their products in gene therapy protocols. Nat Biotechnol 14, 1012–1016 (1996). https://doi.org/10.1038/nbt0896-1012

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