288 Traceless, homology-directed repair-mediated gene editing for junctional epidermolysis bullosa

Abstract

In this study, we used combinations of Cas9 or Cas9 nickase (Cas9n) and single gRNA (sgRNA) ribonucleoprotein (RNP) complexes with single-stranded oligonucleotides (ssODN) to induce homology-directed repair (HDR) in junctional epidermolysis bullosa (JEB) keratinocytes with a pathogenic mutation in exon 52 of COL17A1. To assess the correction efficiency, treated patient keratinocytes were then analysed via next generation sequencing (NGS), Western Blot, sqRT-PCR, Flow cytometry and immunofluorescence stainings of cell monolayers as well as 3D skin equivalents.