Study of electrostatically stabilized nucleopeptide complexes for targeted DNA delivery to muscle cells

Abstract

Genetic constructs must be selectively delivered to target tissues and intracellular compartments in the necessary concentrations to achieve the maximum therapeutic effect in gene therapy. The search for ways to implement targeted non-viral delivery of nucleic acids into cells, including muscle cells, as one of the most difficult to transfect tissues in vivo, remains topical. We have developed coated cationic nucleopeptide complexes containing ASSLNIA ligand for targeted DNA delivery to muscle tissue. We transfected C2C12 myoblasts with subsequent analysis of toxic properties using the Alamar Blue test and detection of lacZ and GFP gene expression. Analysis of DNA compaction by peptide carriers was assessed using the EtBr displacement test. After delivery of plasmid DNA with the GFP gene to the femoral muscle of mdx mice, the presence of fluorescence was assessed using microscopy. Here we show that the developed carriers are non-toxic, can efficiently condense DNA, and protect it from polyanions. The carriers efficiently deliver plasmid DNA to C2C12 myoblasts and m. quadriceps of mdx mice. Results of this study are useful for the development of genetic drugs for the treatment of inherited neuromuscular diseases e.g. Duchenne muscular dystrophy.