Journal article

AAV-Mediated CRISPR/Cas Gene Editing of Retinal Cells In Vivo

Sandy SC Hung, Vicki Chrysostomou, Fan Li, Jeremiah KH Lim, Jiang-Hui Wang, Joseph E Powell, Leilei Tu, Maciej Daniszewski, Camden Lo, Raymond C Wong, Jonathan G Crowston, Alice Pebay, Anna E King, Bang V Bui, Guei-Sheung Liu, Alex W Hewitt

INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE | ASSOC RESEARCH VISION OPHTHALMOLOGY INC | Published : 2016

Abstract

PURPOSE: Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein (Cas) has recently been adapted to enable efficient editing of the mammalian genome, opening novel avenues for therapeutic intervention of inherited diseases. In seeking to disrupt yellow fluorescent protein (YFP) in a Thy1-YFP transgenic mouse, we assessed the feasibility of utilizing the adeno-associated virus 2 (AAV2) to deliver CRISPR/Cas for gene modification of retinal cells in vivo. METHODS: Single guide RNA (sgRNA) plasmids were designed to target YFP, and after in vitro validation, selected guides were cloned into a dual AAV system. One AAV2 construct was used to deliver Streptococc..

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Grants

Funding Acknowledgements

Supported by grants from the BrightFocus Foundation, the Ophthalmic Research Institute of Australia, Retina Australia, the Childhood Eye Cancer Trust, and the Eye Research Australia Fund. AP and BVB are 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.