Targeted Delivery of Plasmid DNA to Cells` Nucleus via Nuclear Localization Signal Peptide Conjugated to Modified Starch Based Carrier - ISMBE 2016 - Biomedical Engineering 2016, International Convention Center Haifa, 24.2.2016

Shachar Gat ¹ Ramesh Chintakunta ¹ Riki Goldbart ¹ Tamar Traitel ¹ Dina Aranovich ¹ Anne Bernheim-Groswasser ^1,2 Joseph Kost ^1,2

¹Chemical Engineering, Ben-Gurion University of the Negev
²Ilse Kats Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev

Transfection with non-viral gene delivery vectors, generally results in low transgene expression in vivo. One of the reasons is poor cytoplasmic transport and intra-nuclear DNA delivery. The import of molecules larger than 45 kDa into the nucleus is mediated by nuclear localization signals (NLS) which recruit import proteins (importins) to their surface as well as motor proteins. Our objective was to design an improved pDNA carrier system that can recruit import and motor proteins and thereby mediate the active movement of the pDNA from the plasma membrane toward the nucleus, docking followed by import to the nucleus through the nuclear pore complex. Starch, a natural polysaccharide was modified into cationic starch (Q-starch) in order to enable self-assembly with the pDNA. Covalent coupling of polyethylene glycol-thiol to the Q-starch was carried out in order to attach NLS peptides. The pDNA delivery system was developed and characterized, specific binding to motor proteins was confirmed. The cellular transfection path was investigated.