engLansing, FelixMukhametzyanova, LiliyaGünes, CerenHoersten, JennaSchmitt, Lukas TheoRodriguez-Muela, NataliaKnöfler, RalfTakebe, TakanoriBuchholz, FrankRojo-Romanos, TeresaIwasawa, KentaroKimura, MasakiPaszkowski-Rogacz, MaciejKarpinski, JanetGrass, TobiasSonntag, JanSchneider, Paul MartinCorrection of a Factor VIII genomic inversion with designer-recombinases.info:eu-repo/classification/ddc/500Amino Acid SequenceBase SequenceCell DifferentiationChromosome Inversion: geneticsClone CellsDirected Molecular EvolutionEndothelial Cells: cytologyEndothelial Cells: metabolismExons: geneticsFactor VIII: geneticsHEK293 CellsHeLa CellsHumansInduced Pluripotent Stem Cells: metabolismInverted Repeat Sequences: geneticsRecombinases: metabolismRecombination, Genetic: geneticsSubstrate SpecificityWhole Genome SequencingRecombinasesFactor VIIIDespite advances in nuclease-based genome editing technologies, correcting human disease-causing genomic inversions remains a challenge. Here, we describe the potential use of a recombinase-based system to correct the 140 kb inversion of the F8 gene frequently found in patients diagnosed with severe Hemophilia A. Employing substrate-linked directed molecular evolution, we develop a coupled heterodimeric recombinase system (RecF8) achieving 30% inversion of the target sequence in human tissue culture cells. Transient RecF8 treatment of endothelial cells, differentiated from patient-derived induced pluripotent stem cells (iPSCs) of a hemophilic donor, results in 12% correction of the inversion and restores Factor VIII mRNA expression. In this work, we present designer-recombinases as an efficient and specific means towards treatment of monogenic diseases caused by large gene inversions.Nature Communications 13(1), 422 (2022). doi:10.1038/s41467-022-28080-7info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionNature Publishing Group UK2022info:eu-repo/semantics/openAccessDEhttps://pub.dzne.de/record/163482https://pub.dzne.de/search?p=id:%22DZNE-2022-00242%22Researchersinfo:eu-repo/semantics/altIdentifier/doi/10.1038/s41467-022-28080-7info:eu-repo/semantics/altIdentifier/issn/2041-1723info:eu-repo/semantics/altIdentifier/pmid/pmid:35058465