Abstract
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein (Cas) is used by some bacteria and most archaea to protect against viral phage intrusion and has recently been adapted to allow for efficient editing of the mammalian genome. Whilst CRISPR/Cas-based technology has been used to modify genes in mammalian cells in vitro, delivery of CRISPR/Cas system into mammalian tissue and/or organs is more difficult and often requires additional vectors. With the use of adeno-associated virus (AAV) gene delivery system, active CRISPR/Cas enzyme can be maintained for an extended period of time and enable efficient editing of genome in the retina in vivo. Herein we outline the method to edit the genome in mouse retina using a dual AAV vector -mediated CRISPR/Cas9 system.
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Acknowledgments
We thank Ian Alexander, Grant Logan, James Bender, Vicki Chrysostomou, Jeremiah Lim, Joseph Powell, Leilei Tu, Maciej Daniszewski, Camden Lo, Raymond Wong, Jonathan Crowston, and Alice Pébay. This work was supported by grants from the National Health and Medical Research Council, the Global Ophthalmology Awards Program, the Ophthalmic Research Institute of Australia, the Royal Hobart Research Foundation, the Childhood Eye Cancer Trust, and the Eye Research Australia Fund. BVB is supported by Australian Research Council Future Fellowships. AWH is supported by a National Health and Medical Research Council Practitioner Fellowship. Centre for Eye Research Australia (CERA) receives operational infrastructure support from the Victorian Government.
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Hung, S.S. et al. (2018). Methods for In Vivo CRISPR/Cas Editing of the Adult Murine Retina. In: Boon, C., Wijnholds, J. (eds) Retinal Gene Therapy. Methods in Molecular Biology, vol 1715. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7522-8_9
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DOI: https://doi.org/10.1007/978-1-4939-7522-8_9
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