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. 2024 Jan 18;15:590. doi: 10.1038/s41467-024-44830-1

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — A Schematic of LNPs encapsulating barcoded DNA (b-DNA) for accelerated in vivo delivery screening. Left: LNPs were formulated via microfluidic mixing of an aqueous phase of b-DNA and an ethanol phase of lipids. Middle: Schematic formation of LNPs encapsulating b-DNA. LNPs were formulated via microfluidic mixing, and each LNP formulation was encapsulated with a unique b-DNA. Right: 15 LNP formulations (B1–B15) were formulated by varying the identity and molar ratio of phospholipid (DOPE, DOTAP, or DSPC) and molar ratio of cholesterol. A 0.5 μg b-DNA dose of each b-DNA-LNP was pooled and administered to C57BL/6 mice intramuscularly as a single injection. 4 h post injection, b-DNA delivery to the heart, liver, spleen, lung, draining lymph nodes, and muscle were quantified. Organs were harvested 4 h post-injection, and b-DNA delivery was quantified using both whole-organ for deep sequencing to identify the top-performing LNP. B Schematic of immunization with LNP-based DNA vaccine and the efficacy in K18-hACE2 mice against SARS-CoV-2 variants. Top: K18-hACE2 mice were immunized with either controls or LNP-HPS and received a booster dose after 3 weeks. Bottom: vaccinated mice displayed decreased lethality post-viral challenge and generated potent cellular and humoral responses against SARS-CoV-2 VOCs, indicating a strong and comprehensive immune response following vaccination. C Darker clusters indicate higher delivery, whereas lighter clusters indicate lower delivery. Within the heatmap, the delivery of different LNP formulations within the same organ (left to right) can be compared. The delivery of the same LNP formulation across different samples (top to bottom) cannot be compared. D, E The bar graph illustrates the percentage quantification of LNP (B1–B15) delivery in different target tissues 4 hours after intramuscular injection. Data are presented as mean ± SEM (n = 4/group). Source data are provided as a Source Data file.