Camilla Brolin Hjortkjær, Denmark
Post doc - Institute of Cellular and Molecular Medicine, University of Copenhagen, Denmark
From April 11th to 15th I visited Professor Matthew Woods’ lab in Oxford on a STSM. I am currently working on a project combining PNA targeting exon 23 with a novel controlled drug release formulation in mdx mice, the animal model of DMD. The main aim of my stay was to learn their quantifications methods for exon 23 skipping in mdx mice both at the RNA and protein level in order to implement their methods in our lab. Another aim was to see and discuss functional testing in mice.
Dr. Graham McClorey showed me their PCR protocols. We discussed RNA extraction method, advantages and disadvantages with nested RT-PCR and RT-qPCR, primer design and we went through data analysis. Dr. Caroline Godfrey and Dr. Corinne Betts talked me through their immunohistochemistry (IHC) protocol and we discussed different IHC dystrophin quantification methods. They showed me their new quantification method using Image pro. Dr. Betts also showed me their western blotting protocol and we discussed loading and quantification of gels. Furthermore, we discussed purification methods for PNA and PMO (HPLC and desalting procedures) and preparation of P-PMOs for i.v. injections. I also spent some time in the animal facility observing and discussing different functional testing in mice with Dr. Melissa Bowerman (rotarod and grid test).
This STSM has been a very inspiring and constructive experience, and will be extremely useful in my future work. In Copenhagen I am the only one in my lab working with exon skipping and DMD, so it was fantastic being in the Wood lab full of experts in field. It resulted in many good discussions and future collaboration. Finally, I would like to thank for Professor Matthew Wood for welcoming me in his lab and to Dr. Corinne Betts for being a great host.