Adenine base editing (ABE) enables enzymatic conversion from A-T into G-C base pairs. ABE holds promise for clinical application, as it does not depend on the introduction of double-strand breaks, contrary to conventional CRISPR/Cas9-mediated genome engineering. Here, we describe a cystic fibrosis (CF) intestinal organoid biobank, representing 664 patients, of which ~20% can theoretically be repaired by ABE. We apply SpCas9-ABE (PAM recognition sequence: NGG) and xCas9-ABE (PAM recognition sequence: NGN) on four selected CF organoid samples. Genetic and functional repair was obtained in all four cases, while whole-genome sequencing (WGS) of corrected lines of two patients did not detect off-target mutations. These observations exemplify the value of large, patient-derived organoid biobanks representing hereditary disease and indicate that ABE may be safely applied in human cells.

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Persistent URL dx.doi.org/10.1016/j.stem.2020.01.019, hdl.handle.net/1765/128468
Journal Cell Stem Cell
Citation
Geurts, M.H., van de Poel, E, Amatngalim, GD, Oka, R., Meijers, F.M., Kruisselbrink, E, … Clevers, H.C. (2020). CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank. Cell Stem Cell, 26(4), 503–50+. doi:10.1016/j.stem.2020.01.019