Plasmid DNA delivery into MDA-MB-453 cells mediated by recombinant Her-NLS fusion protein

Abstract

Sivakumar Jeyarajan*, Jennifer Xavier*, N Madhusudhana Rao, Vijaya GopalCentre for Cellular and Molecular Biology, Council for Scientific and Industrial Research, Hyderabad, Andhra Pradesh, India; *These authors contributed equally to this workAbstract: A major rate-limiting step in nonviral gene delivery is the entry of nucleic acids across various membrane barriers and eventually into the nucleus where it must be transcribed. Cell-penetrating peptides and proteins are employed to generate formulations that overcome these challenges to facilitate DNA delivery into cells efficiently. However, these are limited by their inability to deliver nucleic acids selectively due to lack of specificity because they deliver to both cancer and normal cells. In this study, through modular design, we generated a recombinant fusion protein designated as Her-nuclear localization sequence (Her-NLS), where heregulin-a (Her), a targeting moiety, was cloned in frame with cationic NLS peptide to obtain a cell-specific targeting biomolecule for nucleic acid delivery. The heregulin-a1 isoform possesses the epidermal growth factor-like domain and binds to HER2/3 heterodimers which are overexpressed in certain breast cancers. Purified recombinant Her-NLS fusion protein binds plasmid DNA and specifically transfects MDA-MB-453 cells overexpressing the epidermal growth factor receptors HER2/3 in vitro. The approach described would also permit replacement of heregulin ligand with other targeting moieties that would be suited to cell-specific nucleic acid delivery mediated via receptor-ligand interactions.Keywords: cell targeting, nucleic acid delivery, nuclear localization sequences, heregulin, transfection, HER2 receptors