ALL STEM CELLS

Highlights •CRISPR-Cas9 and targeted sgRNAs can revert large inversions in hemophilia A iPSCs •Endothelial cells derived from corrected express correctly spliced Factor VIII •Transplantation of rescue injury mortality hemophiliac mice •Whole-genome deep sequencing did not detect off-target mutations Summary Hemophilia is an X-linked genetic disorder caused by the F8 gene, which encodes blood coagulation factor VIII. Almost half all severe cases result two gross (140-kbp or 600-kbp) chromosomal that involve introns 1 22 respectively. We induced pluripotent stem (iPSCs) patients with these inversion genotypes used CRISPR-Cas9 nucleases to segments back WT situation. isolated inversion-corrected frequencies up 6.7% without detectable based on whole-genome sequencing. Endothelial differentiated expressed gene functionally rescued deficiency otherwise lethal mouse model hemophilia. Our results therefore provide a proof principle for functional correction rearrangements patient-derived suggest potential therapeutic applications.