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 Powell, Leilei Tu, Maciej Daniszewski, Camden Lo, Raymond Wong, Jonathan Crowston, Alice Pébay, Anna King, Bang Bui, Guei-Sheung Liu, Alex Hewitt

Published : 2016


ABSTRACT PURPOSE CRISPR/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 utilising the adeno-associated virus 2 (AAV2) to deliver CRISPR/Cas for gene modification of retinal cells in vivo . METHODS 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 SpCas9 and the other delivered sgRNA against YFP or LacZ (control) in the presence of mCherry. Five weeks after..

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