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.

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 a 140 kb genomic inversion of the F8 gene, which is frequently found in patients diagnosed with severe Hemophilia A. Employing substrate-linked directed molecular evolution, we developed a fused 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, resulted in prominent correction of the inversion and restored Factor VIII mRNA expression. Our data suggests that designer-recombinases may represent efficient and specific means towards treatment of monogenic diseases caused by large gene inversions.

The paper is devoted to the role of wood in the art of furniture. The work traces the typological relationship of certain types of furniture designed in the twentieth century with the prototypes of the XVIII and XIX centuries. The paper argues about the possibilities of the material for creating aesthetically significant furniture samples. The stage in recognizing the new possibilities of technologically processed wood is the furniture by J. Summers, which revealed the possibility of designing a chair from a single sheet of plywood “in one piece” without structural connections. H. Peak, C. Mollino, Ch. R. Eames, S. Yanagi worked in the same direction. Chipboard is the last of the materials discussed in the paper. The appeal to the archaic box-type design was laid in the works by Rietveld, M. Bill, and G. Gugelot. In the wake of the architecture, deconstruction in the furniture design can be seen in the works by F. and W. Campana.