Cleancap fluc mrna 5mou

Female C57BL/6 (6–8 weeks old) were purchased from the Jackson Laboratories. The mice were housed under specific pathogen-free conditions in the School of Medicine, University of Maryland, Baltimore animal facilities. All animal procedures were performed according to the guidelines and protocols of the University of Maryland, Baltimore Animal Care and use committee and approved by the Institutional Animal Care and use Committee (IACUC; protocol #0721010). Polyplexes were prepared in acetate buffer (25 mM; pH 5) by mixing different polymers (50 mg/mL) and FLuc mRNA (1 mg/mL) (CleanCap^® FLuc mRNA (5moU), TriLink Biotechnologies, San Diego, CA) at 110:1 weight ratio. After the preparation, the polyplexes were dialyzed against DPBS buffer (pH 7.4) for 30–60 minutes to remove DMSO and acetate buffer. All the polymers were stable after dialysis except polyMDET-C10/mRNA. The polyplexes were administered to mice via tail vein injection (10 μg mRNA/injection). After 24 hours, the mice were injected with D-luciferin (300 μL, 15 mg/mL) intraperitoneally. After 15 minutes, the mice were euthanized, and various organs were collected (liver, spleen, heart, lung, kidneys) and imaged using the Xenogen IVIS^® Spectrum Imaging System (Alameda, CA).