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A novel intranuclear RNA vector system for long-term stem cell modification

Gene Therapy volume 23, pages 256–262 (2016)Cite this article

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Abstract

Genetically modified stem and progenitor cells have emerged as a promising regenerative platform in the treatment of genetic and degenerative disorders, highlighted by their successful therapeutic use in inherent immunodeficiencies. However, biosafety concerns over insertional mutagenesis resulting from integrating recombinant viral vectors have overshadowed the widespread clinical applications of genetically modified stem cells. Here, we report an RNA-based episomal vector system, amenable for long-term transgene expression in stem cells. Specifically, we used a unique intranuclear RNA virus, borna disease virus (BDV), as the gene transfer vehicle, capable of persistent infections in various cell types. BDV-based vectors allowed for long-term transgene expression in mesenchymal stem cells (MSCs) without affecting cellular morphology, cell surface CD105 expression or the adipogenicity of MSCs. Similarly, replication-defective BDV vectors achieved long-term transduction of human induced pluripotent stem cells, while maintaining the ability to differentiate into three embryonic germ layers. Thus, the BDV-based vectors offer a genomic modification-free, episomal RNA delivery system for sustained stem cell transduction.

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Acknowledgements

This work was supported by Mayo Foundation, Mayo Clinic Center for Regenerative Medicine, Eisenberg Stem Cell Trust and National Institutes of Health (R01HL098502) to YI; and Funding Program for Next Generation World-Leading Researchers (NEXT program) from Japan Society for the Promotion of Science (JSPS) to KT.

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Authors and Affiliations

  1. Department of Molecular Medicine, Mayo Clinic, Rochester, MN, USA

    Y Ikeda, W E Matchett, S J Holditch & B Lu

  2. Department of Viral Oncology, Institute for Virus Research, Kyoto University, Kyoto, Japan

    A Makino & K Tomonaga

  3. Center for Emerging Virus Research, Institute for Virus Research, Kyoto University, Kyoto, Japan

    A Makino

  4. Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA

    A B Dietz

  5. Department of Tumor Viruses, Graduate School of Medicine, Graduate School of Biostudies, Kyoto University, Kyoto, Japan

    K Tomonaga

  6. Department of Mammalian Regulatory Network, Graduate School of Biostudies, Kyoto University, Kyoto, Japan

    K Tomonaga

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  1. Y Ikeda
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  2. A Makino
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Correspondence to Y Ikeda or K Tomonaga.

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Ikeda, Y., Makino, A., Matchett, W. et al. A novel intranuclear RNA vector system for long-term stem cell modification. Gene Ther 23, 256–262 (2016). https://doi.org/10.1038/gt.2015.108

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