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mRNA Lipoplexes with Cationic and Ionizable α-Amino-lipophosphonates: Membrane Fusion, Transfection, mRNA Translation and Conformation
oleh: Sohail Akhter, Mathieu Berchel, Paul-Alain Jaffrès, Patrick Midoux, Chantal Pichon
Format: | Article |
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Diterbitkan: | MDPI AG 2022-03-01 |
Deskripsi
Cationic liposomes are attractive carriers for mRNA delivery. Here, mRNA lipoplexes (LX) were prepared with the cationic lipids α-aminolipophosphonate (<b>3b</b>) or imidazolium lipophosphoramidate (<b>2</b>) associated with various α-aminolipophosphonates co-lipids comprising protonable groups (imidazole or pyridine) and <b>DOPE</b>. Physicochemical parameters of liposomes and their membrane fusion activity were measured. LXs comprising either <b>3b</b>- or <b>2</b>- allowed transfection of ~25% and 40% of dendritic cells with low cytotoxicity, respectively; the efficiency increased up to 80% when <b>2</b> was combined with the imidazole-based co-lipid <b>1</b>. The transfections were high with <b>3b</b>/<b>1</b>, <b>3b</b>/<b>DOPE</b>, <b>2</b>/<b>1</b> and <b>2</b>/<b>DOPE</b> LXs. We observed that the transfection level was not well correlated with the acid-mediated membrane fusion activity of liposomes supposed to destabilize endosomes. The mRNA release from LXs and its translation capacity after release were studied for the most efficient LXs. The results showed that the more mRNA was condensed, the poorer the translation efficiency after release was. In contrast to DNA, circular dichroism performed on mRNA complexed with <b>2</b>/<b>DOPE</b> revealed the presence of denatured mRNA in LXs explaining this lack of translation efficiency. This is an important parameter that should be stressed for the preparation of mRNA LXs with a conserved mRNA translation activity.