in frame mutation  (ATCC)

Journal: PLoS ONE

Article Title: Antisense PMO cocktails effectively skip dystrophin exons 45-55 in myotubes transdifferentiated from DMD patient fibroblasts

doi: 10.1371/journal.pone.0197084

Figure Lengend Snippet: (A) Schematic diagram of MYOD1 expression vector. (B) Histogram comparison of ZsGreen fluorescence signal vs cell number between healthy and DMD patient fibroblast cells transduced with MYOD1 expression vector. Representative images shown, although results between either patient cell line were similar. (C) Immunocytochemistry of transduced fibroblasts following 18 d (MYOD1), 15 d (MyHC, myosin heavy chain), 18 d (desmin), and 24 d (dystrophin) differentiation, respectively. Pictured are results from a healthy cell line. Nuclei counterstained with DAPI. Scale bars: 100 μm. (D) RT-PCR time-course analysis of dystrophin expression in healthy and patient DMD transdifferentiated fibroblasts. Images are representative, with dystrophin expression observed in all transdifferentiated cell lines.

Article Snippet: Human DMD patient fibroblast cells harboring out-of-frame deletion mutations of dystrophin exons 45–50 (ID: GM05017) and exons 46–50 (ID: GM05162), as well as healthy human fibroblasts (ID: GM23815) were originally obtained in 2012 from the Coriell Institute for Medical Research (Camden, NJ, USA).

Techniques: Expressing, Plasmid Preparation, Comparison, Fluorescence, Transduction, Immunocytochemistry, Reverse Transcription Polymerase Chain Reaction

Journal: PLoS ONE

Article Title: Antisense PMO cocktails effectively skip dystrophin exons 45-55 in myotubes transdifferentiated from DMD patient fibroblasts

doi: 10.1371/journal.pone.0197084

Figure Lengend Snippet: (A) RT-PCR for dystrophin following cocktail PMO transfection in transdifferentiated DMD patient cells. Cells were treated with 1, 3, or 10 μM each PMO. Expected molecular weight of dystrophin exons 45–55 skipped mRNA is 308 bp. (B) Representative immunocytochemistry of transduced DMD fibroblasts following PMO cocktail transfection. Nuclei counterstained with DAPI. Scale bars: 100 μm.

Article Snippet: Human DMD patient fibroblast cells harboring out-of-frame deletion mutations of dystrophin exons 45–50 (ID: GM05017) and exons 46–50 (ID: GM05162), as well as healthy human fibroblasts (ID: GM23815) were originally obtained in 2012 from the Coriell Institute for Medical Research (Camden, NJ, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Transfection, Molecular Weight, Immunocytochemistry