title: A systematic High-Content Screening microscopy approach reveals key roles for Rab33b, OATL1 and Myo6 in nanoparticle trafficking in HeLa cells.
abstract: Synthetic nanoparticles are promising tools for imaging and drug delivery; however the molecular details of cellular internalization and trafficking await full characterization. Current knowledge suggests that following endocytosis most nanoparticles pass from endosomes to lysosomes. In order to design effective drug delivery strategies that can use the endocytic pathway, or by-pass lysosomal accumulation, a comprehensive understanding of nanoparticle uptake and trafficking mechanisms is therefore fundamental. Here we describe and apply an RNA interference-based high-content screening microscopy strategy to assess the intracellular trafficking of fluorescently-labeled polystyrene nanoparticles in HeLa cells. We screened a total of 408 genes involved in cytoskeleton and membrane function, revealing roles for myosin VI, Rab33b and OATL1 in this process. This work provides the first systematic large-scale quantitative assessment of the proteins responsible for nanoparticle trafficking in cells, paving the way for subsequent genome-wide studies.
authors.affiliation: School of Biology and Environmental Science, University College Dublin, Belfield, Dublin 4, Ireland.|Conway Institute for Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Angela
authors.initials: A
authors.lastname: Panarella
authors.affiliation: School of Biology and Environmental Science, University College Dublin, Belfield, Dublin 4, Ireland.|Conway Institute for Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Mariana G
authors.initials: MG
authors.lastname: Bexiga
authors.affiliation: School of Biology and Environmental Science, University College Dublin, Belfield, Dublin 4, Ireland.|Conway Institute for Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: George
authors.initials: G
authors.lastname: Galea
authors.affiliation: School of Biology and Environmental Science, University College Dublin, Belfield, Dublin 4, Ireland.|Conway Institute for Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Elaine D
authors.initials: ED
authors.lastname: O' Neill
authors.affiliation: Centre for BioNano Interactions, School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Anna
authors.initials: A
authors.lastname: Salvati
authors.affiliation: Centre for BioNano Interactions, School of Chemistry and Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Dawson
authors.affiliation: School of Biology and Environmental Science, University College Dublin, Belfield, Dublin 4, Ireland.|Conway Institute for Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
authors.forename: Jeremy C
authors.initials: JC
authors.lastname: Simpson
mesh_terms: D001692:Biological Transport; D003599:Cytoskeleton / Q000378:metabolism; D016503:Drug Delivery Systems; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D015894:Genome, Human; D006367:HeLa Cells; D006801:Humans; D051907:Lysosomal Membrane Proteins / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D008853:Microscopy / Q000379:methods; D018995:Myosin Heavy Chains / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry; D036103:Nanotechnology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism*; D052118:Lysosomal-Associated Membrane Protein 1
chemical_list: D020690:GTPase-Activating Proteins; D051907:Lysosomal Membrane Proteins; D018995:Myosin Heavy Chains; D034741:RNA, Small Interfering; D033921:Vesicular Transport Proteins; D020691:rab GTP-Binding Proteins; C489116:LAMP1 protein, human; D052118:Lysosomal-Associated Membrane Protein 1; C000627575:TBC1D25 protein, human; C093846:early endosome antigen 1; C089785:myosin VI; C486397:RAB33B protein, human