Cholesterol-DP7-ACP-T7 (DAT) (dual-functional peptide on liposome)
Sequence: cholesterol-VQWRIRVAVIRK-ACP-HAIYPRH
siRNA (duplex, 21-mer with 3′ TT overhangs)
| Experiment Id | EXP002001 |
|---|---|
| Paper | A blood–brain barrier- and blood–brain tumor barrier-penetrating siRNA delivery system targeting gli |
| Peptide | Cholesterol-DP7-ACP-T7 (DAT) (dual-functional peptide on liposome) |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | yes |
| Label Confidence | high |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | yes |
| Peptide Concentration | DAT used to modify LNP at 250 μg/mL (chosen as optimal). |
| Rna Concentration | 0.24 μg cy3-siRNA per well (24-well) with 1.44 μg LNP/DAT-LNP (mass ratio 6:1) for uptake/transfection assays; functional assays used DAT-LNP/si slit2 (dose given as 1.2 μg siRNA in migration/invasion pretreatment). |
| Mixing Ratio | DAT-LNP:siRNA mass ratio 6:1 (gel retardation complete at 6:1). |
| Formulation Format | DAT-modified DOTAP:cholesterol liposome (DAT-LNP) electrostatically adsorbing siRNA |
| Formulation Components | DOTAP:cholesterol (1:1 molar; DOTAP 2 mg/mL) liposome + DAT peptide (250 μg/mL for modification; dialysis) + siRNA (complexed at 6:1 mass ratio LNP:siRNA). |
| Size Nm | 130.10 |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | GL261 mouse glioma cells (uptake/transfection, slit2 knockdown, migration/invasion); BV2 microglia (targeting coculture); bEnd.3 brain microvascular endothelial cells (in vitro BBB model); BMDCs/BMDMs (immune modulation assays). |
| Animal Model | |
| Administration Route | |
| Output Type | In vitro uptake + functional RNA effect: siRNA transfection (flow cytometry/microscopy), slit2 mRNA/protein knockdown (RT–PCR/WB), reduced migration/invasion; immune activation (DC maturation, macrophage polarization). |
| Output Value | High transfection of cy3-siRNA in GL261 (flow + microscopy); significant slit2 knockdown; migration/invasion inhibited; DAT-LNP promotes DC maturation and shifts macrophages toward M1 markers (iNOS↑, IL-10↓). |
| Output Units | |
| Output Notes | Uptake pathways: caveolin- and clathrin-mediated endocytosis (CTB/transferrin colocalization; inhibited by genistein/chlorpromazine). |
| Toxicity Notes | DAT-LNP showed slightly lower toxicity vs Lipo2000/LNP in 293T CCK-8 assay (qualitative). |
| Curation Notes |