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Prevention of interferon-stimulated gene expression using microRNA-designed hairpins

Gene Therapy volume 16, pages 142–147 (2009)Cite this article

Abstract

RNA interference allows selective gene silencing, and is widely used for functional analysis of individual genes in vertebrate cells and represents an attractive therapeutic option for treating central nervous system diseases. However, growing evidence exists that the expression of short hairpin RNAs (shRNAs) can trigger cellular immune response resulting in unspecific cellular phenotypes and severe side effects. We found that lentiviral vector (LV)-mediated expression of shRNAs in primary cortical cultures resulted in strong expression of the interferon-stimulated gene oligoadenylate synthetase 1 (Oas1), which was accompanied by accelerated apoptosis and substantial net neuron loss. Modification of the shRNA construct by implementing features of the naturally occurring microRNA-30 (miR-30) precursor avoided Oas1 induction in transduced primary cultures, whereby modification of the passenger strand seems to be a crucial feature to circumvent interferon-stimulated gene expression. This work represents the first experimental study showing that an miR-30-based shRNA construct prevents Oas1 pathway associated off-target effects, which we consider as an essential prerequisite for shRNA use in future gene therapeutic approaches.

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Acknowledgements

We thank Stephanie Schöffmann for excellent technical assistance and Dr Thomas Meitinger for helpful discussions. This work was supported by the NGFN (‘SMP-Human Brain’, FKZ-01GR0449 to MU). and by a KKF grant from the Technical University Munich (KKF-Project 8725151 to MB).

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Authors and Affiliations

  1. Helmholtz Center München-German Research Center for Environmental H,

    M Bauer, N Kinkl, A Meixner & M Ueffing

  2. ealth, Institute of Human Genetics, München-Neuherberg, Germany

    M Bauer, N Kinkl, A Meixner & M Ueffing

  3. Institute of Human Genetics, Klinikum rechts der Isar, TU München, München, Germany,

    M Bauer, N Kinkl, A Meixner & M Ueffing

  4. Department of Neurolog,

    M Bauer & T Gasser

  5. y, Hertie Institute for Clinical Brain Research, University of Tuebingen, Tübingen, Germany

    M Bauer & T Gasser

  6. Helmholtz Center München-German Research Center for Environmental Health, Institute of Molecular Immunology, München, Germany

    E Kremmer

  7. Psychiatrische Klinik, Klinikum rechts der Isar, TU München, München, Germany,

    M Riemenschneider & H Förstl

Authors
  1. M Bauer
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  2. N Kinkl
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  3. A Meixner
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  4. E Kremmer
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  5. M Riemenschneider
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  6. H Förstl
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  7. T Gasser
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  8. M Ueffing
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Correspondence to M Bauer.

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Bauer, M., Kinkl, N., Meixner, A. et al. Prevention of interferon-stimulated gene expression using microRNA-designed hairpins. Gene Ther 16, 142–147 (2009). https://doi.org/10.1038/gt.2008.123

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