Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency

Capel, Victoria and Vllasaliu, Driton and Watts, Peter and Stolnik, Snow (2016) Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency. Biochemical and Biophysical Research Communications, 477 (2). pp. 260-265. ISSN 0006-291X

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Abstract

Despite research efforts, cell uptake processes determining siRNA silencing efficiency remain unclear. Here, we examine the relationship between in vitro cell culture models, cellular trafficking and siRNA silencing efficiency to provide a mechanistic insight on siRNA delivery system design. Model siRNA-polyplexes, based on chitosan as a ‘classical’ condensing agent, were applied to a panel of lung epithelial cell lines, H1299, A549 and Calu-3 and cell internalization levels, trafficking pathways and gene silencing assessed on exposure to pharmacological inhibitors. The data reveal striking differences in the internalization behaviour and gene silencing efficiency in the tested cell lines, despite their common lung epithelial origins. The model system’s silencing was lower where clathrin internalization pathway predominated in Calu-3, relative to silencing in H1299 cells where a non-clathrin internalization appears dominant. Increased silencing on endosomal disruption was apparent in Calu-3 cells, but absent when cellular internalization was not predominantly clathrin-mediated in A549 cells. This highlights that identifying cell trafficking pathways before incorporation of functional components to siRNA delivery systems (e.g. endosomolytic compounds) is crucial. The study hence stresses the importance of selection of appropriate cell culture model, relevant to in vivo target, to assess the gene silencing efficiency and decide which functionalities the ‘stratified siRNA silencing vector’ requires.

Item Type: Article
Keywords: Endocytosis pathways; Lung siRNA therapy; Pharmacological inhibitors; siRNA polyplexes; siRNA silencing
Schools/Departments: University of Nottingham UK Campus > Faculty of Science > School of Pharmacy
Identification Number: https://doi.org/10.1016/j.bbrc.2016.06.054
Depositing User: Eprints, Support
Date Deposited: 22 Jul 2016 16:27
Last Modified: 14 Sep 2016 02:22
URI: http://eprints.nottingham.ac.uk/id/eprint/35333

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