nanoparticle formulations  (Thermo Fisher)

Journal: Frontiers in Molecular Biosciences

Article Title: Efficiency and safety of five different agents for in vivo delivery of novel bioengineered RNAi molecules

doi: 10.3389/fmolb.2026.1785592

Figure Lengend Snippet: Comparative efficacy of BioRNA Gly /GFP-siRNA formulated with five different agents in GFP-transgenic mice in vivo . (A) Schematic representation of the dosing regimen. GFP-transgenic mice (C57BL/6-Tg (CAGEGFP)1Osb/J) were administered iv (via tail vein) with 30 μg of BioRNA Gly /GFP-siRNA every other day for three doses and then sacrificed on day 5, which were delivered in five different ways, including lipid nanoparticle (LNP) made by PackGene Biotech and Invivofectamine (Invivo) from Thermo Fisher Scientific, as well as a Nanoparticle (Nano), LIPID (lipid), and PEG-Liposome (PEG) based In Vivo Transfection Reagents from Altogen Biosystems. (B) GFP siRNA levels in mouse liver tissues were quantified by selective stem-loop RT-qPCR, indicating successful delivery and processing of BioRNA Gly /GFP-siRNA. Notably, GFP siRNA levels in the LNP and Invivofectamine groups were significantly higher than those in the other formulation groups. (C) GFP mRNA levels in mouse livers were determined by RT-qPCR, showing variable degrees of target gene silencing by different formulations. (D) Ex vivo whole-liver fluorescence and (E) frozen liver section imaging demonstrated the variations in suppressing GFP by different formulations. Overall, all formulations offered effective reductions in GFP mRNA levels and fluorescence intensities while the Invivofectamine and LNP formulations led to greater degrees of siRNA accumulation in the liver. Data are mean ± SD. **P < 0.01 and ***P < 0.001 (one-way ANOVA with Bonferroni post hoc tests). Blank: N = 6; LNP-BioRNA Gly /GFP-siRNA: N = 4; Invivo-BioRNA Gly /GFP-siRNA: N = 4; Nano-BioRNA Gly /GFP-siRNA: N = 4; Lipid-BioRNA Gly /GFP-siRNA: N = 3; PEG-BioRNA Gly /GFP-siRNA: N = 3.

Article Snippet: A custom lipid nanoparticle (LNP) formulation was synthesized by PackGene Biotech (Houston, TX, United States), using a DLin-MC3-based lipid composition (the same as Patisiran) to encapsulate BioRNA Gly /GFP-siRNA at a concentration of 0.5 mg/mL, and stored at −80 °C before use.

Techniques: Transgenic Assay, In Vivo, Transfection, Quantitative RT-PCR, Formulation, Ex Vivo, Fluorescence, Imaging

Journal: Frontiers in Molecular Biosciences

Article Title: Efficiency and safety of five different agents for in vivo delivery of novel bioengineered RNAi molecules

doi: 10.3389/fmolb.2026.1785592

Figure Lengend Snippet: Safety profiles of various BioRNA Gly /GFP-siRNA formulations in GFP-transgenic mice. (A) Except an 8%, statistically significant decrease observed in the LNP group, mouse body weights showed no significant differences between any RNA treatment and blank control groups. (B) Weights of mouse major organs, including the lung, kidney, heart, and brain, were comparable between the RNA treatment and blank groups, while LNP product led to an obvious hepatosplenomegaly. Liver weights were also significantly increased by Invivofectamine formulation, while a modest enlargement of spleen is not statistically significant. (C) Mouse blood biochemistry profiles showed that LNP formulation altered the levels of a number of biomarkers, including ALT, AST, albumin, ALP, and total bilirubin, whereas other formulations had no effects. The reference ranges were derived from healthy BALB/c mice and provided by the Comparative Pathology Laboratory at University of California - Davis. Data are mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001 (one- or two-way ANOVA with Bonferroni post hoc tests). Note that only statistically significant pairs are denoted herein, and other unmarked groups are not statistically significant (ns; P > 0.05).

Article Snippet: A custom lipid nanoparticle (LNP) formulation was synthesized by PackGene Biotech (Houston, TX, United States), using a DLin-MC3-based lipid composition (the same as Patisiran) to encapsulate BioRNA Gly /GFP-siRNA at a concentration of 0.5 mg/mL, and stored at −80 °C before use.

Techniques: Transgenic Assay, Control, Formulation, Derivative Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Efficiency and safety of five different agents for in vivo delivery of novel bioengineered RNAi molecules

doi: 10.3389/fmolb.2026.1785592

Figure Lengend Snippet: Influence of different BioRNA Gly /GFP-siRNA formulations on immune responses in GFP-transgenic mice, as manifested by a panel of serum cytokine profiles determined by a Multiplex Cytokine Assay (Eve Technologies). Overall, the LNP formulation remarkably elevated many pro-inflammatory mediators, including MCP-1 (CCL2), MIG (CXCL9), MIP-1β (CCL4), IL-1β, TNFα, RANTES (CCL5), IP-10 (CXCL10), G-CSF, and MIP-2 (CXCL2), compared with the Blank group, indicating strong innate immune activation. In contrast, anti-inflammatory cytokines, including IL-4 and IL-10, did not show any statistically significant differences among treatment groups. Meanwhile, the Invivofectamine and nanoparticle formulations caused mild changes in particular cytokines (e.g., increase in IFNγ by Nano-BioRNA product; and increase in IP-10 and G-CSF by Invivo-BioRNA product), whereas the Lipid and PEG formulations had no or minimal effects. Data are mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001 (one-way ANOVA with Bonferroni post hoc tests). Note that only statistically significant pairs are denoted while other unmarked groups are not statistically significant (ns; P > 0.05).

Article Snippet: A custom lipid nanoparticle (LNP) formulation was synthesized by PackGene Biotech (Houston, TX, United States), using a DLin-MC3-based lipid composition (the same as Patisiran) to encapsulate BioRNA Gly /GFP-siRNA at a concentration of 0.5 mg/mL, and stored at −80 °C before use.

Techniques: Transgenic Assay, Multiplex Assay, Cytokine Assay, Formulation, Activation Assay

Journal: Frontiers in Molecular Biosciences

Article Title: Efficiency and safety of five different agents for in vivo delivery of novel bioengineered RNAi molecules

doi: 10.3389/fmolb.2026.1785592

Figure Lengend Snippet: Comparative efficiency of various BioRNA Gly /GFP-siRNA formulations to reduce GFP intensities in GFP-expressing Huh7 cells in vitro . Cells were transfected with 15 nM of BioRNA Gly /GFP-siRNA, control RNA, or vehicle using the indicated reagents, with Lipofectamine 3000 (Lipo) as a positive control. (A) Representative GFP fluorescence images at 72 h post-transfection. The Invivofectamine ( Invivo ) formulated siRNA significantly reduced GFP fluorescence intensity at 72 h post-transfection, whereas nanoparticle (Nano) and LNP formulated siRNAs did not show much suppression of GFP fluorescence intensities, when compared with control RNA or vehicle. (B) Quantification of GFP fluorescence intensity. Absolute fluorescence intensities (left) and those (right) normalized to respective cell confluency measured by IncuCyte live-cell imaging. (C) Changes in intracellular GFP-siRNA levels in different treatment groups as determined by selective stem-loop RT-qPCR assay. Note that LNP-BioRNA Gly /Control RNA formulation was not prepared for this study, and thus there are no related data. Data are presented as mean ± SD, n = 3. *P < 0.05, **P < 0.01, ***P < 0.001 (one- or two-way ANOVA with Bonferroni post hoc tests). Lipo: Lipofectamine 3000 transfection reagent. Other unmarked groups are not statistically significant (ns; P > 0.05).

Article Snippet: A custom lipid nanoparticle (LNP) formulation was synthesized by PackGene Biotech (Houston, TX, United States), using a DLin-MC3-based lipid composition (the same as Patisiran) to encapsulate BioRNA Gly /GFP-siRNA at a concentration of 0.5 mg/mL, and stored at −80 °C before use.

Techniques: Expressing, In Vitro, Transfection, Control, Positive Control, Fluorescence, Live Cell Imaging, Quantitative RT-PCR, Formulation