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Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo
Journal article   Open access   Peer reviewed

Shuttle peptide delivers base editor RNPs to rhesus monkey airway epithelial cells in vivo

Katarina Kulhankova, Soumba Traore, Xue Cheng, Hadrien Benk-Fortin, Stéphanie Hallée, Mario Harvey, Joannie Roberge, Frédéric Couture, Sajeev Kohli, Thomas J Gross, …
Nature communications, Vol.14(1), 8051
12/05/2023
DOI: 10.1038/s41467-023-43904-w
PMCID: PMC10698009
PMID: 38052872
url
https://doi.org/10.1038/s41467-023-43904-wView
Published (Version of record) Open Access

Abstract

Gene editing strategies for cystic fibrosis are challenged by the complex barrier properties of airway epithelia. We previously reported that the amphiphilic S10 shuttle peptide non-covalently combined with CRISPR-associated (Cas) ribonucleoprotein (RNP) enabled editing of human and mouse airway epithelial cells. Here, we derive the S315 peptide as an improvement over S10 in delivering base editor RNP. Following intratracheal aerosol delivery of Cy5-labeled peptide in rhesus macaques, we confirm delivery throughout the respiratory tract. Subsequently, we target CCR5 with co-administration of ABE8e-Cas9 RNP and S315. We achieve editing efficiencies of up-to 5.3% in rhesus airway epithelia. Moreover, we document persistence of edited epithelia for up to 12 months in mice. Finally, delivery of ABE8e-Cas9 targeting the CFTR R553X mutation restores anion channel function in cultured human airway epithelia. These results demonstrate the therapeutic potential of base editor delivery with S315 to functionally correct the CFTR R553X mutation in respiratory epithelia.Gene editing strategies for cystic fibrosis are challenging. Here the authors improve on their previously reported shuttle peptide noncovalently combined with Cas ribonucleoprotein (RNP), and derive the S315 peptide for delivery: they show base editing in the respiratory tract of the rhesus macaques.
Cystic Fibrosis Ion Channels Mutation Peptides CRISPR Editing Epithelial cells Epithelium Genetic modification Respiratory tract Trachea

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