RT Journal Article
T1 Factor VIII mRNA expression from a BAC carrying the intact locus made by homologous recombination
A1 Pérez Luz, Sara
A1 Abdulrazzak, Hassan
A1 Grillot-Courvalin, Catherine
A1 Huxley, Clare
AB Hemophilia 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.
SN 0888-7543
YR 2007
FD 2007
LK https://hdl.handle.net/20.500.14352/93001
UL https://hdl.handle.net/20.500.14352/93001
LA eng
NO Pérez-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.
NO Ministerio de Educación y Ciencia (España)
NO Medical Research Council (United Kingdom)
NO European Commission 
DS Docta Complutense
RD 9 abr 2025