Unbiased assessment of genome integrity and purging of large deletions at the target locus upon CD4 T-cell CRISPR-Cas9 long range editing for CD40LG deficiency

ABSTRACT: Hyper IgM1 is an X-linked combined immunodeficiency, caused by CD40LG mutations, potentially treatable with CD4+ T-cell Cas9 gene editing with a "one-size-fits-most" corrective template. Contrary to established gene therapies, there is limited data on genomic alterations following long-range gene editing, and no consensus on the relevant assays. We developed drop-off digital PCR assays for unbiased detection of large on-target deletions and found them at high frequency upon editing human cells. Large deletions were common upon editing different loci and cell types and using alternative Cas9 and template delivery methods. In CD40LG edited T cells, on-target deletions were counter-selected in culture and further purged by enrichment for edited cells using a selector coupled to gene correctio

SUBMITTER: Dr. Daniele Canarutto 

PROVIDER: S-SCDT-10_15252-EMBJ_2023114188 | biostudies-other |

REPOSITORIES: biostudies-other