Journal: Nature Communications
Article Title: Engineering a streamlined virus-like particle for programmable tissue-specific gene delivery
doi: 10.1038/s41467-025-64181-9
Figure Lengend Snippet: a Ai9 mice were intravenously injected with 1 ×10 12 gc Cre mRNA containing original VLP (SFV-Env, n = 3) or engineered VLP (Env-P11, n = 4). Scale representing relative levels of fluorescence from high to low (yellow to red). b Liver tdTomato expression was examined by immunofluorescence (tdTomato, green; DAPI, blue; scale bar = 50 μm). c Calculation of the ratio of positive tdTomato cells in different organs. Data represented the mean with standard deviation (SD) of three independent animals and three fields per organ (Brain: PBS vs. SFV-Env, ** P = 0.0068; PBS vs. Env-P11 and SFV-Env vs. Env-P11, **** P < 0.0001. Liver: PBS vs. Env-P11, ** P = 0.0088; PBS vs. SFV-Env and SFV-Env vs. Env-P11, **** P < 0.0001. Spleen: PBS vs. SFV-Env and SFV-Env vs. Env-P11, **** P < 0.0001; two-way ANOVA with Bonferroni’s multiple comparison test). Co-immunofluorescence staining of tdTomato with d neuronal marker NeuN or e glial marker Gfap. (NeuN or Gfap, red; tdTomato, white; DAPI, blue; scale bar = 50 μm). Binding ability of VLP vectors to mouse neural cell lines f N2a (**** P < 0.0001), g bEnd.3 (**** P < 0.0001), and h C8-D1A (**** P < 0.0001), and primary cells i primary pericyte (** P = 0.0046), j primary microvascular endothelial cell (EC) (* P = 0.0429), and k primary astrocyte (*** P = 0.0001), with vector genomic copies per cell of 100,000, normalized using Gapdh . Error bars represented the mean with SD of four independent replicates. Statistical analyses were performed using two-tailed unpaired t test. l Construction of blood-brain barrier spheroids using primary mouse brain microvascular endothelial cells, primary brain microvascular pericytes, and primary astrocytes , . Spheroids were co-incubated with VLPs containing luciferase mRNA at vector genomic copies per cell of m 10,000,000 or n 30,000,000, and luminescence levels of the BBB spheroids were measured. Error bars represented the mean with SD of three independent replicates (* P = 0.0291, two-tailed unpaired t test). Source data are provided as a Source Data file.
Article Snippet: Primary mouse brain microvascular endothelial cells (Procell, CP-M108) were cultured in Mouse Brain Microvascular Endothelial Cell Complete Medium (Procell, CM-M108).
Techniques: Injection, Fluorescence, Expressing, Immunofluorescence, Standard Deviation, Comparison, Staining, Marker, Binding Assay, Plasmid Preparation, Two Tailed Test, Incubation, Luciferase
