Utility of self-destructing CRISPR/Cas constructs for targeted gene editing in the retina

F Li; SSC Hung; MKN Mohd Khalid; JH Wang; V Chrysostomou; VHY Wong; V Singh; K Wing; L Tu; JA Bender; A Pébay; AE King; AL Cook; RCB Wong; BV Bui; AW Hewitt; GS Liu

Journal article

Utility of self-destructing CRISPR/Cas constructs for targeted gene editing in the retina

F Li, SSC Hung, MKN Mohd Khalid, JH Wang, V Chrysostomou, VHY Wong, V Singh, K Wing, L Tu, JA Bender, A Pébay, AE King, AL Cook, RCB Wong, BV Bui, AW Hewitt, GS Liu

Human Gene Therapy | MARY ANN LIEBERT, INC | Published : 2019

DOI: 10.1089/hum.2019.021

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Abstract

Safe delivery of CRISPR/Cas endonucleases remains one of the major barriers to the widespread application of in vivo genome editing. We previously reported the utility of adeno-associated virus (AAV)-mediated CRISPR/Cas genome editing in the retina; however, with this type of viral delivery system, active endonucleases will remain in the retina for an extended period, making genotoxicity a significant consideration in clinical applications. To address this issue, we have designed a self-destructing "kamikaze" CRISPR/Cas system that disrupts the Cas enzyme itself following expression. Four guide RNAs (sgRNAs) were initially designed to target Streptococcus pyogenes Cas9 (SpCas9) and after in ..

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University of Melbourne Researchers

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Awarded by Bayer

Funding Acknowledgements

This work was supported by funding from a Bayer Global Ophthalmology Award, the Ophthalmic Research Institute of Australia, an Australian National Health and Medical Research Council (NHMRC) grant (no. 1123329), an NHMRC Practitioner Fellowship (A.W.H., no. 1103329), an NHMRC Senior Research Fellowship (A.P., no. 1154389), and an Australian Research Council Future Fellowship (A.P., FT140100047).