Exon skipping therapy using antisense oligonucleotides

Gisela Gaina, Romania

PhD

Short term scientific mission (STSM) – From generation of primary muscle culture to exon skipping therapy using antisense oligonucleotides.
Gisela Gaina, Romania

Department of Biochemistry and Molecular Biology, University of Bucharest, Romania

My Short Term Scientific Mission took place from the 14th of November to the 2nd of December 2016 at the Neuromuscular Disorders Group led by Dr Virginia Arechavala-Gomeza BioCruces Health Research Institute, Barakaldo, Spain.

The aim of my STSM was to acquire new knowledge about lab techniques associated with exon skipping therapy using antisense oligonucleotides, and also in the generation of primary cultures from biopsies.

During the three weeks of the training I had the opportunity to learn and to gain hands-on experience in cell culture techniques: both from a patient with muscular dystrophy and also in culture initiation from frozen vials, the maintenance, cell transfection, differentiation/quantification assays and storage of cultures in liquid nitrogen.

I was also trained in lab techniques associated with exon skipping therapy using antisense oligonucleotides, beginning with design of AONs, cell transfection using Lipofectamine 2000, RNA extraction and cDNA synthesis followed by exon skipping evaluation by two-round RT-PCR.

In all this period I had the opportunity to see and to learn methods used in Neuromuscular Disorders Group like oligo annealing and cloning into backbone vectors.

The STSM was very fruitful experience for me and gave me the opportunity to learn from experimented researchers. The knowledge and experience that I gained at Neuromuscular Disorders Group lab is of utmost value for my current work and also for future projects and I am truly grateful especially to Dr. Virginia Arechavala-Gomeza as well as the rest of the her team.