A NOVEL SPT ANALYZER SHEDS LIGHT ON NLS-COATED NANO-PARTICLES MOVING IN LIVE CELLS - ISBE 2015 Conference

Yael Lavi ¹ Levi A. Gheber ^1,2 Dina Aranovitch ³ Anne Bernheim ³

¹Avram and Stella Godstein-Goren Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva
²Ilse Katz Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev, Beer-Sheva
³Department of Chemical Engineering, Ben-Gurion University of the Negev, Beer-Sheva

Nuclear protein transport is vital to many cell processes, as well as to those pathogens that seek to infect and exploit cell resources and machinery. Understanding the modes of transports of Nuclear Localization Signal (NLS)-coated nano-particles (NLS-NP) in living cells gives insight into this important mechanism. This may give rise to tools for manipulating or harnessing this mechanism to further the field of targeted drug/therapeutic genes delivery.

Thousands of NLS-NP tracks were analyzed with our newly developed SPT analyzer. Through comparing such parameters as progression vector in relation to the nucleus, turning angle, track duration, confinement ratio, displacement in time, start and end points and distance from the nucleus, it is possible to analyze hundreds of tracks simultaneously and infer sub-populations with regard to mode of diffusion. Assisted diffusion is expected since the NLS tag recruits dynein motors onto microtubules to traffic attached NPs to the nucleus.

Our study shows coating NPs in NLS has significant effect on their movement inside living cells, which is attenuated by both the concentration and the distribution of NLS across the particles. NLS coating even has significant effect on the NPs ability to enter cells.

Understanding Nuclear protein transport allows designing novel drugs that are organelle specific. Specific targeting increases drug efficiency, significantly lowering dosages and drug toxicity.