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Efficient marking of neural stem cell-derived neurons with a modified murine embryonic stem cell virus, MESV2

Gene Therapy volume 9, pages 1044–1048 (2002)Cite this article

Abstract

Treatments for nervous system disorders that involve transplanting genetically modified neural stem cells may ultimately be feasible. As a step towards this therapeutic approach, a novel murine embryonic stem cell gammaretroviral vector was developed with features designed to optimize transgene expression in neural stem cells and to increase vector safety. All potential start sites of translation in the 5’ leader were removed. These sites may compete with an inserted transgene for translation initiation, and also produce potentially immunogenic peptides. Further, all of the gag gene sequences were replaced with a well-defined constitutive transport element from avian leukemia virus to promote nuclear export of viral RNA, and to eliminate any homology between the vector and a murine leukemia virus-derived gag-pol packaging plasmid. Two versions of the virus were made in which EGFP expression was driven either by the Rous sarcoma virus U3 enhancer or by a combination of sequences from the Syn1 and Pgk-1 promoters. Both of these viruses efficiently transduced neural stem cells isolated from embryonic rat hippocampus, and robust EGFP expression was observed in neurons derived from these cells following differentiation in vitro.

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Acknowledgements

We thank Anna Tran for excellent technical assistance. Flow cytometry was carried out at The Scripps Research Institute's core facility. Funding for this work was provided by US Public Health Service Grant HD09635 (GME), the Neurosciences Support Corporation, and the G Harold and Leila Mathers Charitable Foundation.

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Authors and Affiliations

  1. The Neurosciences Institute, San Diego, CA, USA

    GC Owens & GM Edelman

  2. Department of Neurobiology, The Scripps Research Institute, La Jolla, CA, USA

    S Mistry, GM Edelman & KL Crossin

  3. The Skaggs Institute for Chemical Biology, La Jolla, CA, USA

    GM Edelman

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Owens, G., Mistry, S., Edelman, G. et al. Efficient marking of neural stem cell-derived neurons with a modified murine embryonic stem cell virus, MESV2. Gene Ther 9, 1044–1048 (2002). https://doi.org/10.1038/sj.gt.3301780

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