Comparison of Gene-Editing Approaches for Severe Congenital Neutropenia-Causing Mutations in the <i>ELANE</i> Gene.

ABSTRACT: Safety considerations for gene therapies of inherited preleukemia syndromes, including severe congenital neutropenia (CN), are paramount. We compared several strategies for CRISPR/Cas9 gene editing of autosomal-dominant ELANE mutations in CD34⁺ cells from two CN patients head-to-head. We tested universal and allele-specific ELANE knockout, ELANE mutation correction by homology-directed repair (HDR) with AAV6, and allele-specific HDR with ssODN. All strategies were not toxic, had at least 30% editing, and rescued granulopoiesis in vitro. In contrast to published data, allele-specific indels in the last exon of ELANE also restored granulopoiesis. Moreover, by implementing patient-derived induced pluripotent stem cells for GUIDE-Seq off-target ana

SUBMITTER: Ritter MU 

PROVIDER: S-EPMC12344121 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature