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Genospheres: self-assembling nucleic acid-lipid nanoparticles suitable for targeted gene delivery

Abstract

We describe the assembly of a cationic lipid-nucleic acid nanoparticle from a liquid monophase containing water and a water miscible organic solvent where both lipid and DNA components are separately soluble prior to their combination. Upon removal of the organic solvent, stable and homogenously sized (70–100 nm) lipid-nucleic acid nanoparticles (Genospheres™) were formed. The low accessibility (<15%) of the nanoparticle-encapsulated DNA to a DNA intercalating dye indicated well-protected nucleic acids and high DNA incorporation efficiencies. It was demonstrated that Genospheres could be stably stored under a variety of conditions including a lyophilized state where no appreciable increase in particle size or DNA accessibility was observed following reconstitution. Finally, Genospheres were made target-specific by insertion of an antibody-lipopolymer (anti-HER2 scFv (F5)-PEG-DSPE) conjugate into the particle. The target specificity (>100-fold) in HER2 overexpressing SK-BR-3 breast cancer cells was dependent on the degree of PEGylation, where the incorporation of high amounts of PEG-lipid on the particle surface (up to 5 mol%) had only a minor effect on the transfection activity of the targeted Genospheres. In summary, this work describes a novel, readily scalable method for preparing highly stable immunotargeted nucleic acid delivery vehicles capable of achieving a high degree of specific transfection activity.

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Acknowledgements

We thank Drs Joel Cohen and Valentina Khorosheva of the University of the Pacific, San Francisco for the zeta potential measurements. We also would like to thank Dr Brigitte Papahadjopoulos-Sternberg for performing the FFEM.

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Correspondence to K Hong.

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Hayes, M., Drummond, D., Kirpotin, D. et al. Genospheres: self-assembling nucleic acid-lipid nanoparticles suitable for targeted gene delivery. Gene Ther 13, 646–651 (2006). https://doi.org/10.1038/sj.gt.3302699

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  • DOI: https://doi.org/10.1038/sj.gt.3302699

Keywords

  • nonviral gene delivery
  • plasmid encapsulation
  • targeted gene delivery
  • HER2 receptor

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