Pérez Luz, SaraAbdulrazzak, HassanGrillot-Courvalin, CatherineHuxley, Clare2024-01-152024-01-152007Pérez-Luz, Sara, et al. «Factor VIII mRNA Expression from a BAC Carrying the Intact Locus Made by Homologous Recombination». Genomics, vol. 90, n.o 5, noviembre de 2007, pp. 610-19. https://doi.org/10.1016/j.ygeno.2007.07.005.0888-754310.1016/j.ygeno.2007.07.005https://hdl.handle.net/20.500.14352/93001Hemophilia A is caused by mutations in the gene encoding factor VIII (F8) and is an important target for gene therapy. The F8 gene contains 26 exons spread over ∼186 kb and no work using the intact genomic locus has been carried out. We have constructed a 250-kb BAC carrying all 26 exons, the introns, and more than 40 kb of upstream and 20 kb of downstream DNA. This F8 BAC was further retrofitted with either the oriP/EBNA-1 elements from Epstein–Barr virus, which allow episomal maintenance in mammalian cells, or alphoid DNA, which allows human artificial chromosome formation in some human cell lines. Lipofection of the oriP/EBNA-1-containing version into mouse Hepa1-6 cells resulted in expression of F8 mRNA spanning the F8 gene. The >300-kb BAC carrying alphoid DNA was successfully delivered to 293A and HT1080 cells using bacterial delivery, resulting in greater than endogenous levels of F8 mRNA expression.engAttribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/journal articlehttps://doi.org/10.1016/j.ygeno.2007.07.005restricted access636.09577Hemophilia AFactor VIIIF8Gene therapyBacterial artificial chromosomeBACBacterial deliveryOriP/EBNA-1Alphoid DNABiologíaVeterinaria2415 Biología Molecular3109 Ciencias Veterinarias