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Factors governing the transduction efficiency of adeno-associated virus in the retinal ganglion cells following intravitreal injection

Gene Therapy volume 26, pages 109–120 (2019)Cite this article

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Abstract

Efficient transduction of the retinal ganglion cells (RGCs) is a prerequisite to maximize therapeutic outcomes in any form of gene therapy for optic neuropathies. Whereas subretinal injection of adeno-associated virus 2 (AAV2) has been well-characterized, the serotype, viral load, and promoter combinations that govern RGC transduction efficiency following intravitreal injection remains poorly understood. We evaluated the transduction efficiency of seven AAV2 serotypes (AAV2/1, AAV2/2, AAV2/4, AAV2/5, AAV2/6, AAV2/8, and AAV2/9) for the RGCs at 4 weeks following intravitreal injection in C57BL/6J mice. Intravitreal injection of 1 × 109 vg of AAV2/2 with eGFP driven by the CMV promoter attained a higher transduction efficiency for the RGCs (60.0 ± 4.2%) compared with the six other AAV2 serotypes with eGFP driven by the same promoter injected at the same viral load ( < 3.0%). Reporter driven by the CAG promoter had a lower transduction efficiency (up to 42.0 ± 5.8%) compared with that driven by the CMV reporter (60.0 ± 4.2%, p ≤ 0.024). There was a viral dose-dependent transduction effect of AAV2/2-CMV-eGFP and the transduction efficiency was 40.2 ± 3.9%, 16.6 ± 4.2%, and 2.6 ± 0.2% when the viral load decreased to 5 × 108 vg, 1 × 108 vg, and 1 × 107 vg, respectively. Optimizing viral serotype, viral load, and promoter construct of AAV2 is important to maximize transgene expression in RGC-targeted gene therapy.

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Acknowledgements

This study was supported by funding from the Hong Kong Research Grant Council General Research Fund 2015/2016 (14101215).

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  1. These authors contributed equally: Xu Cao, Jasmine Yung

Authors and Affiliations

  1. Department of Ophthalmology and Visual Sciences, The Chinese University of Hong Kong, Hong Kong, People’s Republic of China

    Xu Cao, Jasmine Yung, Heather Mak & Christopher K. S. Leung

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  1. Xu Cao
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Correspondence to Christopher K. S. Leung.

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Cao, X., Yung, J., Mak, H. et al. Factors governing the transduction efficiency of adeno-associated virus in the retinal ganglion cells following intravitreal injection. Gene Ther 26, 109–120 (2019). https://doi.org/10.1038/s41434-019-0060-0

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