Generation of embryonic stem cells and mice for duchenne research

PLoS Curr. 2013 Sep 10; doi: 10.1371/


Duchenne muscular dystrophy (DMD) is a muscle-wasting disease in which muscle is continuously damaged, resulting in loss of muscle tissue and function. Antisense-mediated exon skipping is a promising therapeutic approach for DMD. This method uses sequence specific antisense oligonucleotides (AONs) to reframe disrupted dystrophin transcripts. As AONs function in a sequence specific manner, human specific AONs cannot be tested in the mdx mouse, which carries a mutation in the murine Dmd gene. We have previously generated a mouse model carrying the complete human DMD gene (hDMD mouse) integrated in the mouse genome to overcome this problem. However, as this is not a disease model, it cannot be used to study the effect of AON treatment on protein level and muscle function. Therefore, our long term goal is to generate deletions in the human DMD gene in a mouse carrying the hDMD gene in an mdx background. Towards this aim, we generated a male ES cell line carrying the hDMD gene while having the mdx point mutation. Inheritance of the hDMD gene by the ES cell was confirmed both on DNA and mRNA level. Quality control of the ES cells revealed that the pluripotency marker genes Oct-4 and Nanog are well expressed and that 85% of cells have 40 chromosomes. Germ line competence of this cell line has been confirmed, and 2 mice strains were derived from this cell line and crossed back on a C57BL6 background: hDMD/mdx and mdx(BL6).

Grant support

MV and LV are paid by a grant from the Prinses Beatrix Spierfonds (the Netherlands) and AAR receives funding from ZonMw, the Prinses Beatrix Spierfonds and the Duchenne Parent Project (the Netherlands). The infrastructure of the Center for Medical Systems Biology and the Center for Biomedical Genetics was utilized to conduct the reported work. JK, JC, AAR, and JV report being employed by LUMC, which has patents on exon skipping. As a co-author on some of these patents AAR is entitled to a share of royalties. MV, JK, JC, LV and JV declare no conflict of interest.