REpair of heterozygous Mutations independent of Exogenous Donor template with high efficiency (REMEDY) using allele specific CRISPR targeting and HDR enhancers
Year: 2025
Type: preprint
Abstract: Summary We report the development of REMEDY (REpair of heterozygous Mutations independent of Exogenous Donor template with high efficiencY), a genome editing strategy that allows efficient repair of heterozygous mutations in human and mouse cells without necessitating an exogenous donor DNA template. Here, we used in-PAM or near-PAM CRISPR strategies to induce a double-strand break (DSB) in mutant alleles. Following the DSB, the wild-type homologous chromosome itself serves as an endogenous DNA donor template and initiates the correction of the mutant allele. Concurrently treating the cells with HDR enhancers, such as AZD7648, further improved the efficiency of the correction. We demonstrated the utility of REMEDY in the context of six different diseases with heterozygous mutations such as IBMPFD, cystic fibrosis, progeria, ITPR3-associated combined immunodeficiency, ACTA1, and TBCD in human patient derived primary cells and complementary mouse model cell lines. Abstract Figure Graphical Abstract
Authors: Yasemin Sezgin, Genesis Snyder, Noushin Saljoughian, Colin Maguire, Ebru Erzurumluoglu Gokalp, Devi Jaganathan, Eleonora S D’Ambrosio, Burcak Ozes, Gregory Wheeler, Ben Kelly, Mark Hester, Zarife Sahenk, Sriram Vaidyanathan, Afrooz Rashnonejad, Jerry Mendell, Nizar Y. Saad, Roshini S. Abraham, Juhi Bagaitkar, Susan D. Reynolds, Allison Bradbury, Meisam Naeimi Kararoudi
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W4410879430