Transfection efficiency of lipoplexes for site-directed delivery

Reint K. Jellema, Paul Bomans, Niko Deckers, Liset Ungethum, Chris P. M. Reutelingsperger, Leo Hofstra, Peter M. Frederik*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Targeted gene delivery is a promising strategy to cure disease on its basic level at the site of interest. The ultrastructure, internalization, and transfection efficiency of lipoplexes was investigated. We found that at a charge ratio (rho) of 4.0 lipoplexes had optimum characteristics for gene delivery in vitro. To decrease the size of lipoplexes, we used a method of continuous-flow microfluidics. PEGylation of lipoplexes did not hinder internalization, but was found to hamper transfection. To discriminate between uptake and transfection efficiency of lipoplexes, we used fluorescence-based approaches: microscopy and FACS. To this end, GFP plasmid was labeled with Alexa 594, and, in parallel experiments, GFP plasmid was combined with rhodamine-labeled lipid. Our studies confirm that cellular uptake does not imply transfection efficiency, and that hurdles in cellular processing have to be taken before targeted gene delivery becomes an established therapeutic option.
Original languageEnglish
Pages (from-to)258-267
JournalJournal of Liposome Research
Volume20
Issue number3
DOIs
Publication statusPublished - Sept 2010

Keywords

  • Gene delivery
  • lipoplexes
  • PEGylation
  • microfluidics
  • transfection efficiency

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