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Reprogramming axonal behavior by axon-specific viral transduction

Gene Therapy volume 19, pages 947–955 (2012)Cite this article

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Abstract

The treatment of axonal disorders, such as diseases associated with axonal injury and degeneration, is limited by the inability to directly target therapeutic protein expression to injured axons. Current gene therapy approaches rely on infection and transcription of viral genes in the cell body. Here, we describe an approach to target gene expression selectively to axons. Using a genetically engineered mouse containing epitope-labeled ribosomes, we find that neurons in adult animals contain ribosomes in distal axons. To use axonal ribosomes to alter local protein expression, we utilized a Sindbis virus containing an RNA genome that has been modified so that it can be directly used as a template for translation. Selective application of this virus to axons leads to local translation of heterologous proteins. Furthermore, we demonstrate that selective axonal protein expression can be used to modify axonal signaling in cultured neurons, enabling axons to grow over inhibitory substrates typically encountered following axonal injury. We also show that this viral approach also can be used to achieve heterologous expression in axons of living animals, indicating that this approach can be used to alter the axonal proteome in vivo. Together, these data identify a novel strategy to manipulate protein expression in axons, and provides a novel approach for using gene therapies for disorders of axonal function.

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Acknowledgements

We thank S Schlesinger and A Jeromin for Sindbis plasmids, Joe Harris (UC Irvine), for preparing masters for casting microfluidic chambers, MS Cohen, A Deglincerti, L Levin and J Buck (Weill Cornell Medical College) for useful comments and suggestions. This work was supported by the Christopher Reeve Paralysis Foundation, the New York State Spinal Cord Injury Board, NIH Grants NINDS NS56306 (SRJ), DA08259 (TAM), ML096571 (TAM), ML098351 (TAM), the Biomembrane Plasticity Research Center (2011-0000841) through the National Research Foundation funded by the Ministry of Education, Science and Technology (NLJ) and training grant T32DA007274 (BAW).

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Author notes

  1. U Hengst

    Present address: 6Present address: The Taub Institute for Research on Alzheimer's Disease and the Aging Brain, Department of Pathology and Cell Biology, Columbia University, New York, NY 10032, USA.,

Authors and Affiliations

  1. Department of Pharmacology, Weill Medical College, Cornell University, New York, NY, USA

    B A Walker, U Hengst & S R Jaffrey

  2. Division of WCU Multiscale Mechanical Design, School of Mechanical and Aerospace Engineering, Seoul National University, Seoul, Korea

    H J Kim & N L Jeon

  3. Laboratory of Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, NY, USA

    E F Schmidt & N Heintz

  4. Division of Neurobiology, Department of Neurology and Neuroscience, Weill Medical College, Cornell University, New York, NY, USA

    T A Milner

  5. Harold and Margaret Miliken Hatch Laboratory of Neuroendocrinology, The Rockefeller University, New York, NY, USA

    T A Milner

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Correspondence to S R Jaffrey.

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Walker, B., Hengst, U., Kim, H. et al. Reprogramming axonal behavior by axon-specific viral transduction. Gene Ther 19, 947–955 (2012). https://doi.org/10.1038/gt.2011.217

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