Extended Data Fig. 7: Knockdown of Itgav or Itgb3 reduces the lung delivery of 6R-LNP in vivo.
(a) Percentage of VtnR+ cells in mouse heart, liver, spleen, lung, and kidney, respectively. n = 3 animals for each condition. (b) Experiment design of EGFP mRNA delivery in C57/BL6 wild-type mice using 6R-LNP. (c) Quantification of VtnR+ and VtnR- cell populations, respectively, within EGFP+ cells in the lung after EGFP mRNA delivery by 6R-LNP. n = 3 animals for each condition. (d) Experiment design for the knockdown of Itgav and/or Itgb3 in the lung of C57/BL6 wild-type mice using CRISPR/Cas9 system with 6R-LNP. (e) qPCR analysis of the knockdown of Itgav and/or Itgb3 expression in the lung under indicated conditions. n = 3 animals for each condition. (f) Flow cytometry analysis of the knockdown of Itgav and/or Itgb3 receptor expression in the lung under indicated conditions. n = 3 animals for each condition. (g) Quantification of Itgav+Itgb3+ (that is, VtnR + ) cell population in the lung after gene editing under indicated conditions. n = 3 animals for each condition. (h) The workflow of Luc mRNA delivery using 6R-LNP. (i) Ex vivo organ luminescence images showing the expression level of luciferase in the liver, lung, spleen, heart, and kidney, respectively, of mice with lung-edited knockdown of Itgav and/or Itgb3 as indicated. n = 3 animals for each condition. (j) Quantification of luminescence of the lung of gene-edited mice as indicated, after Luc mRNA delivery by 6R-LNP. n = 3 animals for each condition. One-way analysis of variance (ANOVA) and Dunnett’s multiple comparisons test were used for statistical analysis. ns: p > 0.05. All data were plotted as the mean ± s.d. Schematics in b, d and h created using BioRender.com.
