Functional correction of CFTR mutations in human airway epithelial cells using adenine base editors |
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| Authors: | Sateesh Krishnamurthy Soumba Traore Ashley L Cooney Christian M Brommel Katarina Kulhankova Patrick
L Sinn Gregory
A Newby David
R Liu Paul
B McCray Jr |
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| Institution: | Department of Pediatrics, University of Iowa, Iowa City, IA, USA;Molecular Medicine Graduate Program, Pappajohn Biomedical Institute, University of Iowa, Iowa, City, IA, USA;Merkin Institute of Transformative Technologies in Healthcare, Broad Institute of Harvard and MIT, Cambridge, MA, USA;Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA;Howard Hughes Medical Institute, Harvard University, Cambridge, MA, USA |
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| Abstract: | Mutations in the CFTR gene that lead to premature stop codons or splicing defects cause cystic fibrosis (CF) and are not amenable to treatment by small-molecule modulators. Here, we investigate the use of adenine base editor (ABE) ribonucleoproteins (RNPs) that convert A•T to G•C base pairs as a therapeutic strategy for three CF-causing mutations. Using ABE RNPs, we corrected in human airway epithelial cells premature stop codon mutations (R553X and W1282X) and a splice-site mutation (3849 + 10 kb C > T). Following ABE delivery, DNA sequencing revealed correction of these pathogenic mutations at efficiencies that reached 38–82% with minimal bystander edits or indels. This range of editing was sufficient to attain functional correction of CFTR-dependent anion channel activity in primary epithelial cells from CF patients and in a CF patient-derived cell line. These results demonstrate the utility of base editor RNPs to repair CFTR mutations that are not currently treatable with approved therapeutics. |
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