Complex Size and Surface Charge Determine Nucleic Acid Transfer by Fusogenic Liposomes

• Published in: International journal of molecular sciences 2020-03, Vol.21 (6), p.2244
• Main Authors: Hoffmann, Marco, Hersch, Nils, Gerlach, Sven, Dreissen, Georg, Springer, Ronald, Merkel, Rudolf, Csiszár, Agnes, Hoffmann, Bernd
• Format: Article
• Language: English
• Subjects: Animals; Biocompatibility; Biomedical materials; Cell culture; Cell cycle; Cell fusion; Charge transfer; CHO Cells; Cricetinae; Cricetulus; Cytoplasm; Deoxyribonucleic acid; DNA; Efficiency; Endocytosis; fusogenic liposomes; Interaction parameters; Lipids; Liposomes; Liposomes - chemistry; Mammalian cells; Membrane Fusion; mRNA; Nanoparticles; nucleic acid complexation; Nucleic acids; Nucleic Acids - chemistry; Nucleic Acids - genetics; Plasma; Plasmids; Polymers; Static Electricity; Surface charge; Transfection; Transfection - methods; Transfection - standards
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21062244

Highly efficient, biocompatible, and fast nucleic acid delivery methods are essential for biomedical applications and research. At present, two main strategies are used to this end. In non-viral transfection liposome- or polymer-based formulations are used to transfer cargo into cells via endocytosis, whereas viral carriers enable direct nucleic acid delivery into the cell cytoplasm. Here, we introduce a new generation of liposomes for nucleic acid delivery, which immediately fuse with the cellular plasma membrane upon contact to transfer the functional nucleic acid directly into the cell cytoplasm. For maximum fusion efficiency combined with high cargo transfer, nucleic acids had to be complexed and partially neutralized before incorporation into fusogenic liposomes. Among the various neutralization agents tested, small, linear, and positively charged polymers yielded the best complex properties. Systematic variation of liposomal composition and nucleic acid complexation identified surface charge as well as particle size as essential parameters for cargo-liposome interaction and subsequent fusion induction. Optimized protocols were tested for the efficient transfer of different kinds of nucleic acids like plasmid DNA, messenger RNA, and short-interfering RNA into various mammalian cells in culture and into primary tissues.