A toolkit to study nuclear transport pathways: nanobody-mediated inhibition of importin β-related nuclear transport receptors (NTRs)

Abstract

Intro -- Thesis Committee and Examination Board Members -- Epigraph -- List of Figures -- List of Tables -- Summary -- 1 Introduction -- 1.1 Compartmentalization of eukaryotic cells -- 1.2 Nuclear pore complex -- 1.3 Permeability barrier of the NPC -- 1.4 Nuclear transport receptors -- 1.5 Overview of selected NTRs -- 1.6 Tools to investigate NTR mediated transport pathways -- 1.7 Nanobodies -- 1.8 Aim of this study -- 2 Results -- 2.1 Biochemical characterization of anti-NTR nanobodies -- 2.2 In vivo validation of Nb-mediated NTR pathway inhibition -- 2.3 Inhibition of NTR-facilitated transport through NPCs inpermeabilized cells -- 2.4 Tag-Nbs to interrupt NTR-facilitated transport through NPCsin permeabilized cells -- 2.5 Impeding NTR-facilitated transport into reconstituted FGhydrogels -- 3 Discussion -- 3.1 Functional classification of the selected anti-NTR Nbs -- 3.2 Tag-Nbs: the good, the bad, and the redundant -- 3.3 Future Outlook -- 4 Materials and Methods -- 4.1 Chemicals, proteins, and reagents -- 4.2 Molecular cloning -- 4.3 Protein purification -- 4.4 Binding assays -- 4.5 In vivo experiments -- 4.6 Permeabilized cell assays -- 4.7 Experiment using FG phase hydrogels -- References -- Acknowledgements.