Rowan Asfahani, PhD

This job involved combining gene and stem cell therapy for Duchenne Muscular Dystrophy. The objective of this work was to develop a therapeutic lentiviral vector carrying the dystrophin gene and transducing this into the most effective primary muscle stem cell to produce high expression of dystrophin. Over the course of the three years, my day-to-day laboratory duties included the maintenance of numerous cell lines including both primary murine and human cell lines, which I derived from transgenic mice and humans with mutations in the Dystrophin gene. Working with these tools, I was able to develop valuable skills which included RNA extraction and cDNA synthesis following myofibre isolation which was also facilitated by flow cytometry analysis. Myopathic phenotypes were characterized for stem cell characteristics and phenocopied by cellular transfections employing lentiviral vectors coding for dystrophin gene variants. Furthermore, I was responsible for genotyping several colonies of transgenic mouse models for the entire unit, which greatly facilitated my skills for team-work and independent project management.