sars-cov-2 spike(b.1.617.2) gene orf cdna clone expression plasmid  (Sino Biological)

Journal: bioRxiv

Article Title: Polymer-lipid hybrid nanoparticle enhances mRNA delivery and T cell-mediated immunity

doi: 10.64898/2026.01.22.701138

Figure Lengend Snippet: (a) Schematic illustration of the design of LNP and PLNP. (b) Left: sizes of LNP and PLNP measured by dynamic light scattering (DLS). Right: TEM images of LNP and PLNP. Scale bars: 100 nm. (c) Polydispersity index (PDI) of LNP and PLNP. (d) mRNA encapsulation efficiencies of LNP and PLNP. (e) Spike mRNA translation in RAW 264.7 macrophages after incubation with spike mRNA-loaded LNP or PLNP for 48 hours. Spike expression were measured by flow cytometry. Dose: 1 µg mRNA/mL. n = 3. (f) FLuc mRNA translation in RAW 264.7 macrophages on day 1, 2, 3, and 5 after incubation with FLuc mRNA-loaded LNP or PLNP. Dose: 1 µg mRNA/mL. n = 3. The FLuc signals were normalized to the day-1 signal of LNP-treated group. (g) Left: Representative flow plots of CD11c + DCs expressing SIINFEKL/H-2K b after incubation with OVA mRNA-loaded LNP or PLNP for 48 hours. Right: The percentages of CD11c + SIINFEKL/H-2K b+ DCs in total DCs. Dose: 1 µg mRNA/mL. n = 3. (h) Intracellular cytokine levels in RAW 264.7 macrophages after incubation with spike mRNA-loaded LNP, spike mRNA-loaded PLNP, empty LNP, or empty PLNP for 24 hours. Left: IFN-γ. Middle: TNF-α. Right: IL-2. Dose: 1 µg mRNA/mL. n = 3. (i) Cytokine secretion from human PBMCs after incubation with spike mRNA-loaded LNP, spike mRNA-loaded PLNP, empty LNP, or empty PLNP for 24 hours. Dose: 1 µg mRNA/mL. n = 3. (j) Schematic illustration of PLNP enhancing antigen presentation and proinflammatory cytokine generation in APCs. Data are presented as mean ± s.e.m. Statistical significance was determined using two-sided unpaired t-test (d, e, g), one-way ANOVA with Tukey post hoc test for multiple comparisons (h), or two-way ANOVA with Tukey post hoc test for multiple comparisons ns: no significance, * P < 0.05, ** P < 0.01, *** P < 0.001, and ****P < 0.0001.

Article Snippet: Wild-type SARS-CoV-2 spike encoding plasmid was obtained from the MacMaster lab. Omicron spike (B.1.1.529), influenza A H1N1 (A/Puerto Rico/8/1934) hemagglutinin (HA), and FLuc sequences were obtained from commercially available omicron spike, HA, and FLuc Gene ORF cDNA clone expression plasmids (Sino Biological #VG40835-UT, Sino Biological #VG11684-UT, and Addgene #101156) and inserted into the same plasmid backbone using Gibson assembly.

Techniques: Encapsulation, Incubation, Expressing, Flow Cytometry, Immunopeptidomics

Journal: bioRxiv

Article Title: Polymer-lipid hybrid nanoparticle enhances mRNA delivery and T cell-mediated immunity

doi: 10.64898/2026.01.22.701138

Figure Lengend Snippet: (a) Left: In vivo luminescence images of BALB/c mice treated with 1×PBS, FLuc mRNA-loaded LNP, or FLuc mRNA-loaded PLNP at 4, 24, 48, 72, and 96 hours post administration. Right: In vivo luminescence signals measured by integration of total flux for each mouse. Dose: 5 µg mRNA/mouse. N = 5. (b) Left: Fluorescence images of lymph nodes collected from C57BL/6 mice at 24 h post vaccination with DiD-labeled spike mRNA-loaded LNP or PLNP. Right: Quantification of LNP and PLNP accumulation in lymph nodes at 24 h post vaccination. Dose: 10 µg mRNA/mouse. n = 4. (c) Innate immune cell activation (CD86 + ) at 7 days post boost vaccination. C57BL/6 mice were vaccinated with spike mRNA-loaded LNP or PLNP on day 0 and boosted with same doses on day 21. Dose: 10 µg mRNA/mouse. n = 5. Left: DCs. Middle: Macrophages. Right: Monocytes. (d) Schematic illustration of PLNP enhancing lymph node targeting and innate immune cell activation. Data are presented as mean ± s.e.m. Statistical significance was determined using two-way ANOVA with Tukey post hoc test for multiple comparisons (a) or one-way ANOVA with Tukey post hoc test for multiple comparisons (b, c). ns: no significance, * P < 0.05, ** P < 0.01, *** P < 0.001, and ****P < 0.0001.

Article Snippet: Wild-type SARS-CoV-2 spike encoding plasmid was obtained from the MacMaster lab. Omicron spike (B.1.1.529), influenza A H1N1 (A/Puerto Rico/8/1934) hemagglutinin (HA), and FLuc sequences were obtained from commercially available omicron spike, HA, and FLuc Gene ORF cDNA clone expression plasmids (Sino Biological #VG40835-UT, Sino Biological #VG11684-UT, and Addgene #101156) and inserted into the same plasmid backbone using Gibson assembly.

Techniques: In Vivo, Fluorescence, Labeling, Activation Assay