iPS-CNM: an iPSC collection generated by base editing for centronuclear myopathy

Abstract

Centronuclear myopathy (CNM) is a rare form of inherited diseases often caused by single base mutations. Modelling CNM is challenging due to the diversity of CNM mutations and genetic background of individual patient. To address this, we used base editing to introduce CNM mutations into an induced pluripotent stem cell (iPSC) line from a healthy donor, generating a collection of iPSC lines (iPS-CNM) carrying distinct CNM mutations. We found that the efficiency of base editing depended critically on selecting base editor (BE) variants and the target sequences. Optimization using different BE variant and sgRNA pair was required for each target site. Moreover, whole genome sequencing (WGS) was performed to confirm on-targets and detect off-targets in order to select iPSC clones for the collection. Our findings highlight the feasibility of base editing for generating an iPSC collection from one parental iPSC line combined with thorough evaluation of rare off-targets using WGS.