Journal article

Comparison of CRISPR/Cas Endonucleases forin vivoRetinal Gene Editing

Fan Li, Kristof Wing, Jiang-Hui Wang, Chi D Luu, James A Bender, Jinying Chen, Qi Wang, Qinyi Lu, Minh Thuan Nguyen Tran, Kaylene M Young, Raymond CB Wong, Alice Pebay, Anthony L Cook, Sandy SC Hung, Guei-Sheung Liu, Alex W Hewitt

Frontiers in Cellular Neuroscience | FRONTIERS MEDIA SA | Published : 2020

Abstract

CRISPR/Cas has opened the prospect of direct gene correction therapy for some inherited retinal diseases. Previous work has demonstrated the utility of adeno-associated virus (AAV) mediated delivery to retinal cells in vivo; however, with the expanding repertoire of CRISPR/Cas endonucleases, it is not clear which of these are most efficacious for retinal editing in vivo. We sought to compare CRISPR/Cas endonuclease activity using both single and dual AAV delivery strategies for gene editing in retinal cells. Plasmids of a dual vector system with SpCas9, SaCas9, Cas12a, CjCas9 and a sgRNA targeting YFP, as well as a single vector system with SaCas9/YFP sgRNA were generated and validated in YF..

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Grants

Awarded by Australian National Health and Medical Research Council (NHMRC)


Awarded by NHMRC


Funding Acknowledgements

This work was supported by funding from a Bayer Global Ophthalmology Award, The Ophthalmic Research Institute of Australia, the Royal Hobart Hospital Research Foundation, an Australian National Health and Medical Research Council (NHMRC) grant (GNT1123329), an NHMRC Practitioner Fellowship (AH, GNT1103329), a NHMRC Career Development Award (KY, GNT1045240), and an NHMRC Research Fellowship NHMRC Senior Research Fellowship (AP, GNT1154389). Centre for Eye Research Australia receives Operational Infrastructure Support from the Victorian Government.