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. 2026 Apr 23;17:5648. doi: 10.1038/s41467-026-72203-3

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© The Author(s) 2026

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 1 — a Schematic depicting the chimeric antigen receptor constructs. b Flow chart of in vitro mRNA transcription. Created in BioRender. Zhao, K. (2024) https://BioRender.com/s23r279. c TEM imaging of the CD34/LNP-CAR mRNA; and (d, e) DLS analysis of the CD34/LNP-CAR mRNA. Data in (c) are representative images from two independent experiments. f Time-dependent DLS size in PBS for 7 days. Data in (f) are from n = 3 biologically independent samples. g Workflow for CD34⁺ HSPCs isolation. Created in BioRender. Zhao, K. (2024) https://BioRender.com/x57e354. h Cell surface markers identified by flow cytometric analysis. Data in (h) are from n = 3 biologically independent samples. i Confocal images of CD34⁺ HSPCs treated with PBS, Fr-RNAs, LNP-CAR mRNAs, or CD34/LNP-CAR mRNA. mRNA was labeled with Cy5 (red), and the cell membrane and nuclei were counterstained with DIO (green) and DAPI (blue), respectively. j Cellular uptake of CD34/LNP-CAR mRNA by CD34⁺ HSPCs via flow cytometry analysis. Data in (j) are from n = 3 biologically independent samples. k Confocal images of subcellular compartments of CD34⁺ HSPCs incubated with CD34/LNP-CAR mRNA for 1, 3, and 6 h at 37 °C. l Quantitative analysis of co-localization of Cy5-mRNAs with endo/lysosomes labeled with Lysotracker Green. The cell nuclei were stained using Hoechst 33342 (blue), endo/lysosomes were stained using Lysotracker Green (green), and mRNAs were labeled with Cy5 (red). Data in (l) are from n = 3 biologically independent samples. m Viability of CD34⁺ HSPCs in different treatment groups at 24 h post-treatment. Data in (m) are from n = 5 biologically independent samples. All data are presented as mean ± s.d. Statistical significance and P-values of (j), and (m) were analyzed by one-way ANOVA followed by Tukey’s multiple comparisons test (two-sided). Statistical significance and P- values of (l) were analyzed by two-way ANOVA followed by Tukey’s multiple comparisons test (two-sided). *P < 0.05, **P < 0.01, ***P < 0.001. Scale bars, 200 or 100 nm (c); 10 μm (i); 10 μm (k). Source data are provided as a Source Data file.