Influence of Lipid Composition of Cationic Liposomes 2X3-DOPE on mRNA Delivery into Eukaryotic Cells

Author:

Vysochinskaya Vera,Shishlyannikov SergeyORCID,Zabrodskaya YanaORCID,Shmendel Elena,Klotchenko SergeyORCID,Dobrovolskaya OlgaORCID,Gavrilova NinaORCID,Makarova Darya,Plotnikova Marina,Elpaeva Ekaterina,Gorshkov Andrey,Moshkoff Dmitry,Maslov MikhailORCID,Vasin AndreyORCID

Abstract

The design of cationic liposomes for efficient mRNA delivery can significantly improve mRNA-based therapies. Lipoplexes based on polycationic lipid 1,26-bis(cholest-5-en-3β-yloxycarbonylamino)-7,11,16,20-tetraazahexacosane tetrahydrochloride (2X3) and helper lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) were formulated in different molar ratios (1:1, 1:2, 1:3) to efficiently deliver model mRNAs to BHK-21 and A549. The objective of this study was to examine the effect of 2X3-DOPE composition as well as lipid-to-mRNA ratio (amino-to-phosphate group ratio, N/P) on mRNA transfection. We found that lipoplex-mediated transfection efficiency depends on both liposome composition and the N/P ratio. Lipoplexes with an N/P ratio of 10/1 showed nanometric hydrodynamic size, positive ζ potential, maximum loading, and transfection efficiency. Liposomes 2X3-DOPE (1:3) provided the superior delivery of both mRNA coding firefly luciferase and mRNA-eGFP into BHK-21 cells and A549 cells, compared with commercial Lipofectamine MessengerMax.

Funder

Ministry of Health of the Russian Federation

the Ministry of Science and Higher Education of the Russian Federation under the strategic aca-demic leadership program “Priority 2030”

Publisher

MDPI AG

Subject

Pharmaceutical Science

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