17 Lansing, Felix Mukhametzyanova, Liliya Rojo-Romanos, Teresa Iwasawa, Kentaro Kimura, Masaki Paszkowski-Rogacz, Maciej Karpinski, Janet Grass, Tobias Sonntag, Jan Schneider, Paul Martin Günes, Ceren Hoersten, Jenna Schmitt, Lukas Theo Rodriguez-Muela, Natalia Knöfler, Ralf Takebe, Takanori Buchholz, Frank AG Rodriguez-Muela Nature Communications Nature Communications Nature Publishing Group UK [London] 2041-1723 10.1038/s41467-022-28080-7 English 422 1 13 Despite 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. PUB:(DE-HGF)16, ; 0, ; Amino Acid Sequence Base Sequence Cell Differentiation Chromosome Inversion: genetics Clone Cells Directed Molecular Evolution Endothelial Cells: cytology Endothelial Cells: metabolism Exons: genetics Factor VIII: genetics HEK293 Cells HeLa Cells Humans Induced Pluripotent Stem Cells: metabolism Inverted Repeat Sequences: genetics Recombinases: metabolism Recombination, Genetic: genetics Substrate Specificity Whole Genome Sequencing Recombinases Factor VIII Journal Article 2022 DZNE-2022-00242 2022 pmc:PMC8776779 pmid:35058465 https://pub.dzne.de/record/163482 https://doi.org/10.1038/s41467-022-28080-7