Gene-Delivery Ability of New Hydrogenated and Partially Fluorinated Gemini bispyridinium Surfactants with Six Methylene Spacers |
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| Authors: | Michele Massa Mirko Rivara Gaetano Donofrio Luigi Cristofolini Erica Peracchia Carlotta Compari Franco Bacciottini Davide Orsi Valentina Franceschi Emilia Fisicaro |
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| Affiliation: | 1.Department of Maternal Infantile and Urological Sciences, Sapienza University of Rome, 00165 Rome, Italy;2.Department Food and Drug, University of Parma, Parco Area delle Scienze 27/A, 43124 Parma, Italy; (M.R.); (E.P.); (C.C.); (F.B.);3.Department of Veterinary Sciences, University of Parma, Via del Taglio 10, 43126 Parma, Italy; (G.D.); (V.F.);4.Department of Mathematical, Physical and Computer Science, University of Parma, Parco Area delle Scienze 7/a, 43124 Parma, Italy; (L.C.); (D.O.) |
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| Abstract: | The pandemic emergency determined by the spreading worldwide of the SARS-CoV-2 virus has focused the scientific and economic efforts of the pharmaceutical industry and governments on the possibility to fight the virus by genetic immunization. The genetic material must be delivered inside the cells by means of vectors. Due to the risk of adverse or immunogenic reaction or replication connected with the more efficient viral vectors, non-viral vectors are in many cases considered as a preferred strategy for gene delivery into eukaryotic cells. This paper is devoted to the evaluation of the gene delivery ability of new synthesized gemini bis-pyridinium surfactants with six methylene spacers, both hydrogenated and fluorinated, in comparison with compounds with spacers of different lengths, previously studied. Results from MTT proliferation assay, electrophoresis mobility shift assay (EMSA), transient transfection assay tests and atomic force microscopy (AFM) imaging confirm that pyridinium gemini surfactants could be a valuable tool for gene delivery purposes, but their performance is highly dependent on the spacer length and strictly related to their structure in solution. All the fluorinated compounds are unable to transfect RD-4 cells, if used alone, but they are all able to deliver a plasmid carrying an enhanced green fluorescent protein (EGFP) expression cassette, when co-formulated with 1,2-dioleyl-sn-glycero-3-phosphoethanolamine (DOPE) in a 1:2 ratio. The fluorinated compounds with spacers formed by six (FGP6) and eight carbon atoms (FGP8) give rise to a very interesting gene delivery activity, greater to that of the commercial reagent, when formulated with DOPE. The hydrogenated compound GP16_6 is unable to sufficiently compact the DNA, as shown by AFM images. |
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| Keywords: | heterocyclic gemini cationic surfactants nonviral vectors gene delivery gene therapy partially fluorinated gemini surfactants atomic force microscopy on DNA DNA-surfactant interaction |
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