Enhanced target cell specificity and uptake of lipid nanoparticles using RNA aptamers and peptides (2021)
Sequence: H-HAIYPRH-NH2
RNA aptamer + Cy5 DNA probe (aptamer–probe hybrid)
| Experiment Id | EXP000985 |
|---|---|
| Paper | Enhanced target cell specificity and uptake of lipid nanoparticles using RNA aptamers and peptides |
| Peptide | T7 |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | yes |
| Label Confidence | high |
| In Vitro Functional Effect | no |
| Endosomal Escape Evidence | |
| Peptide Concentration | postinsertion: T7-lipid to 3.0 µg/mL; 18 µL added to 90 µL LNP; 30 min at 25°C, 250 rpm |
| Rna Concentration | aptamer–probe: 1.4 nmol in 30 µL PBS added to 165 µL LNP; DNA/lipid ~0.05 w/w; final ~3 mg/mL lipid in PBS |
| Mixing Ratio | LNPs:cells = 1000:1 (transwell uptake; macrophage cytokines; viability) |
| Formulation Format | LNPs loaded with aptamer–Cy5 DNA probe hybrids; Tat/T7 added by postinsertion |
| Formulation Components | DLin-MC3-DMA/DSPC/Cholesterol/DMG-PEG2000 (0.4:0.1:0.4:0.1 mol fraction); G-3 (CCR5) RNA aptamer + Cy5 DNA probe (Cy5/TCTATCAATCTATCA); T7 lipopeptide (postinserted) |
| Size Nm | 66.50 |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro (BBB transwell + uptake; immunogenicity; viability) |
| Cell Lines Or Primary Cells | hCMEC/D3 transwell BBB model; HeLa; TZM-bl (CCR5+); HEK293T; HEK293T-gp160; human MDMs |
| Animal Model | |
| Administration Route | |
| Output Type | in vitro uptake/BBB transit (FACS Cy5 % positive + MFI); immunogenicity; viability |
| Output Value | G-3 (CCR5) aptamer increased uptake into CCR5+ TZM-bl vs HeLa; Tat/T7 did not improve BBB penetration over aptamer alone. |
| Output Units | |
| Output Notes | aptamer + peptide. Uptake quantified by flow cytometry (Cy5) after 24 h exposure in BBB transwell model (hCMEC/D3 TEER ~30 Ω·cm2) and by direct addition; macrophage uptake by fluorescence microscopy; cytokines by Luminex; viability by alamarBlue. |
| Toxicity Notes | HeLa: ~20% reduction in viability with aptamer-loaded LNPs (A-1 or G-3) and with peptide-only (Tat or T7); no additional loss with aptamer+peptide combos. HEK293T: no notable loss. Primary macrophages: no cytokine induction vs PBS. |
| Curation Notes |