Julienne Dietz (Witten / DE; Bochum / DE), Frank Jacobsen (Bochum / DE), Heidi Zhuge (Bochum / DE), Nassam Daya (Bochum / DE), Kamel Mamchaoui (Paris / FR), Wenli Zhang (Witten / DE), Anja Ehrhardt (Witten / DE), Prof. Dr. Matthias Vorgerd (Bochum / DE), Eric Ehrke-Schulz (Witten / DE)
Abstract-Text (inkl. Referenzen)
Background Depending on the therapy approach and disease background, the heterogeneity of muscular tissues complicates the development of targeted gene therapy, where either expression in all muscle types or restriction to only one muscle type is warranted. Muscle specificity can be achieved using promotors mediating tissue specific and sustained physiological expression in the desired muscle types but limited activity in non-targeted tissue. Several muscle specific promotors have been described, but direct comparisons between them are lacking. Here we present a direct comparison of muscle specific DES, MHCK7, miRNA206 and CAPN3 promoter.
Materials/Methods To directly compare the muscle specific promotors, we utilized transfection of reporter plasmids using an in vitro model based on electrical pulse stimulation (EPS) to provoke sarcomere formation in 2D cell culture for quantification of promoter activities in differentiated mouse and human myotubes.
Results We found that DES and MHCK7 promoters showed stronger reporter gene expression levels in proliferating and differentiated myogenic cell lines than miR206 and CAPN3 promoter. However, DES and MHCK7 promotor promoted gene expression also in cardiac cells whereas miR206 and CAPN3 promoter expression was restricted to skeletal muscle.
Conclusion Our results provide direct comparison of muscle specific promotors regarding expression strengths and specificity as this is an important feature for the desired therapy approach.