C12-200 is an ionizable cationic lipidoid commonly utilized as a nucleic acid delivery vehicle[1]. C12-200 possesses unique ionization properties, enabling it to bind with negatively charged nucleic acid molecules, such as mRNA, under physiological conditions to form stable complexes. This facilitates efficient nucleic acid encapsulation and promotes cytosolic delivery, making it applicable in drug delivery, vaccine development, and gene therapy, particularly suited for mRNA vaccine delivery systems[2-4].
In vitro, pretreatment of rat dorsal root ganglion (DRG) neurons, rat primary cortical neurons, and the human cortical neuron cell line (HCN-2) with C12-200 lipid nanoparticles (LNPs) at 50nM for 4–24 hours significantly enhanced siRNA cellular uptake efficiency while maintaining high cell viability[5]. Co-incubation of ARPE-19 cells and MIO-M1 Müller glial cells with C12-200 LNPs (loaded with 1μg mRNA or 0.2–1μg saRNA) for 24 hours increased enhanced green fluorescent protein (eGFP) expression levels in the cells[6].
In vivo, administration of C12-200 LNPs (1mg mRNA/kg) via intraperitoneal or intravenous injection in 6-8-week-old female C57BL/6 mice significantly altered the biodistribution pattern of mRNA. Compared to the cholesterol-based benchmark LNP (S2), C12-200 LNPs exhibited strong liver tropism[7]. Additionally, intraperitoneal injection of C12-200 LNPs (1mg/kg siRNA-Cy5.5) in 8-12-week-old Balb/c mice significantly enhanced the uptake of Cy5.5-labeled siRNA in GATA6+ large peritoneal macrophages (LPMs) and induced the migration of LPMs to the lung tissue via systemic circulation within 12-24 hours following clodronate-mediated depletion of alveolar macrophages (AMs)[8].
References:
[1] Reinhart AG, Osterwald A, Ringler P, et al. Investigations into mRNA Lipid Nanoparticles Shelf-Life Stability under Nonfrozen Conditions. Mol Pharm. 2023 Dec 4;20(12):6492-6503.
[2] Barbieri BD, Peeler DJ, Samnuan K, et al. The role of helper lipids in optimising nanoparticle formulations of self-amplifying RNA. J Control Release. 2024 Oct;374:280-292.
[3] Kauffman KJ, Dorkin JR, Yang JH, et al. Optimization of Lipid Nanoparticle Formulations for mRNA Delivery in Vivo with Fractional Factorial and Definitive Screening Designs. Nano Lett. 2015 Nov 11;15(11):7300-6.
[4] Labonia MCI, Estapé Senti M, et al. Cardiac delivery of modified mRNA using lipid nanoparticles: Cellular targets and biodistribution after intramyocardial administration. J Control Release. 2024 May;369:734-745.
[5] Khare P, Dave KM, Kamte YS, et al. Development of Lipidoid Nanoparticles for siRNA Delivery to Neural Cells. AAPS J. 2021 Dec 6;24(1):8.
[6] Li W, Vanluchene H, Raes L, et al. Efficacy versus immunogenicity of LNP-mediated delivery of mRNA and self-amplifying RNA upon intravitreal injection in the mouse eye. J Control Release. 2025 Sep 10;385:114027.
[7] Patel SK, Billingsley MM, Mukalel AJ, et al. Bile acid-containing lipid nanoparticles enhance extrahepatic mRNA delivery. Theranostics. 2024 Jan 1;14(1):1-16.
[8] Oza D, Ivich F, Pace J, et al. Lipid nanoparticle encapsulated large peritoneal macrophages migrate to the lungs via the systemic circulation in a model of clodronate-mediated lung-resident macrophage depletion. Theranostics. 2024 Apr 8;14(6):2526-2543.