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A compact dual promoter adeno-associated viral vector for efficient delivery of two genes to dorsal root ganglion neurons

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Abstract

Adeno-associated viral (AAV) vectors based on serotype 5 are an efficient means to target dorsal root ganglia (DRG) to study gene function in the primary sensory neurons of the peripheral nervous system. In this study, we have developed a compact AAV dual promoter vector composed of the cytomegalovirus (CMV) and chicken beta-actin (CAG) promoters in a back-to-back configuration with a shared enhancer, and show efficient expression of two proteins simultaneously in DRG neurons. We demonstrate how this is useful for experiments on axonal regeneration, by co-expressing a gene of interest and an axonal marker. Using a farnesylated form of eGFP, which is actively transported along axons, we show superior long-distance labelling of axons of DRG neurons compared with normal eGFP. Additionally, we have efficiently transduced lumbar DRG neurons by injecting the AAV dual promoter vector into the dorsal intrathecal space, which is a less invasive delivery method. In summary, we have developed an AAV dual promoter vector designed for simultaneous expression of a gene of interest and a fluorescent protein to label long-distance axonal projections, which allows specific quantification of axons from transduced neurons after injury.

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Acknowledgements

This study was partially funded by grants from the International Spinal Reseach Trust (Strategy Grant STR111) and the Netherlands Organisation for Scientific Research (NWO-grant 40-00812-98-110).

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Authors and Affiliations

  1. Laboratory for Neuroregeneration, Netherlands Institute for Neuroscience, an Institute of the Royal Academy of Arts and Sciences, Amsterdam, The Netherlands

    N D Fagoe, R Eggers, J Verhaagen & M R J Mason

  2. Center for Neurogenomics and Cognition Research, Neuroscience Campus Amsterdam, Vrije Universtiteit Amsterdam, Amsterdam, The Netherlands

    J Verhaagen

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  1. N D Fagoe
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  2. R Eggers
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  4. M R J Mason
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Correspondence to M R J Mason.

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This work was carried out in Amsterdam, The Netherlands.

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Fagoe, N., Eggers, R., Verhaagen, J. et al. A compact dual promoter adeno-associated viral vector for efficient delivery of two genes to dorsal root ganglion neurons. Gene Ther 21, 242–252 (2014). https://doi.org/10.1038/gt.2013.71

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