Journal article
Screening of CRISPR/cas base editors to target the AMD high-risk Y402H complement factor H variant
MTN Tran, MKNM Khalid, A Pébay, AL Cook, HH Liang, RCB Wong, JE Craig, GS Liu, SS Hung, AW Hewitt
Molecular Vision | MOLECULAR VISION | Published : 2019
Abstract
Purpose: To evaluate the efficacy of using a CRISPR/Cas-mediated strategy to correct a common high-risk allele that is associated with age-related macular degeneration (AMD; rs1061170; NM_000186.3:c.1204T>C; NP_000177.2:p. His402Tyr) in the complement factor H (CFH) gene. Methods: A human embryonic kidney cell line (HEK293A) was engineered to contain the pathogenic risk variant for AMD (HEK293A-CFH). Several different base editor constructs (BE3, SaBE3, SaKKH-BE3, VQR-BE3, and Target-AID) and their respective single-guide RNA (sgRNA) expression cassettes targeting either the pathogenic risk variant allele in the CFH locus or the LacZ gene, as a negative control, were evaluated head-to-head f..
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Grants
Awarded by Macular Disease Foundation Australia
Funding Acknowledgements
This work was supported by the Ophthalmic Research Institute of Australia and the Macular Disease Foundation of Australia. Financial support was also obtained from an Australian National Health and Medical Research Council (NHMRC) Centres of Research Excellence (CRE) #1023911 and Project Grant (APP1123329). JEC and AWH are supported by NHMRC Fellowships, while AP is supported by an ARC Future Fellowship. The Centre for Eye Research Australia (CERA) receives Operational Infrastructure Support from the Victorian Government. The contents of the published material are solely the responsibility of the Administering Institution, a Participating Institution or individual authors and do not reflect the views of the NHMRC. We gratefully thank Vikrant Singh for helping with the matlab scripts for base calling and indel formation.