Targeted genomic inversion using zinc finger nucleases : Toward gene therapy for Haemophilia A

Abstract

As whole genome sequencing technology develops rapidly, there are many reports about structural variation in genome. Structural variations, which include insertions, deletions, duplications, translocations and inversions, occur in more wide region of genome than single-nucleotide polymorphisms. These structural variations are found in normal populations of human in some cases, but these are the cause of many genetic diseases. Especially, Haemophilia A and Hunter syndrome are caused by genomic inversion. For this reasons, structural variations are highly regarded in the fields of therapy of genetic disorders. Despite the importance of structural variations, it is difficult to investigate the mechanism of these, because there are no molecular tools to induce structural variations in genome. Here in this study, inducements of genomic inversions in human genomes are attempted using zinc finger nuclease technology. Zinc finger nucleases are made to induce genomic inversions in the region of Intron 1 of F8 gene-cause of severe Haemophilia A-and validated by various methods. Theses results hold new promises for the gene therapy of Haemophilia A and it is hoped that zinc finger nuclease technologies are widely used in the fields of gene therapy, based on this study.