CRISPR-Based Adenine Editors Correct Nonsense Mutations in a Cystic Fibrosis Organoid Biobank

Maarten H Geurts, Eyleen de Poel, Gimano D Amatngalim, Rurika Oka, Fleur M Meijers, Evelien Kruisselbrink, Peter van Mourik, Gitte Berkers, Karin M de Winter-de Groot, Sabine Michel, Danya Muilwijk, Bente L Aalbers, Jasper Mullenders, Sylvia F Boj, Sylvia W F Suen, Jesse E Brunsveld, Hettie M Janssens, Marcus A Mall, Simon Y Graeber, Ruben van BoxtelCornelis K van der Ent, Jeffrey M Beekman, Hans Clevers

Onderzoeksoutput: Bijdrage aan wetenschappelijk tijdschrift/periodieke uitgaveArtikelWetenschappelijkpeer review

Samenvatting

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.

Originele taal-2Engels
Pagina's (van-tot)503-510.e7
TijdschriftCell Stem Cell
Volume26
Nummer van het tijdschrift4
DOI's
StatusGepubliceerd - 02 apr 2020

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