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.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:91150 |
| Date | 30 May 2024 |
| Creators | 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 |
| Publisher | Nature Publishing Group |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:article, info:eu-repo/semantics/article, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 2041-1723, 422, 10.1038/s41467-022-28080-7, info:eu-repo/grantAgreement/European Commission/H2020 | ERC | ERC-ADG/742133//Designer recombinases for efficient and safe genome surgery/GENSURGE |
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