3TAT-DRBD (within STD fusion protein)
Sequence: GRKKRRQRRRGSHGRKKRRQRRRGSHGRKKRRQRRRGSHAGDLSAGFFMEELNTYRQKQGVVLKYQELPNSGPPHDRRFTFQVIIDGREFPEGEGRSKKEAKNAAAKLAVEILNKEKKA
| Experiment Id | EXP001477 |
|---|---|
| Paper | A specific aptamer-cell penetrating peptides complex delivered siRNA efficiently and suppressed pros |
| Peptide | 3TAT-DRBD (within STD fusion protein) |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | yes |
| Label Confidence | high |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | |
| Peptide Concentration | Complex dosing: 200 pmol sur-siRNA per 5×10^5 cells; final siRNA 100–400 nM; STD used at ~4–10× siRNA molar in assembly (example 20–50 µM STD with 5 µM siRNA during prep). |
| Rna Concentration | In vitro: 200 pmol siRNA per 5×10^5 cells (final 100–400 nM depending on volume); confocal/flow assays used 200 pmol FAM-sur-siRNA. |
| Mixing Ratio | Theoretical assembly: A10:STD:siRNA = 4:4:1 (4 DRBD per siRNA; 1 SA per biotin-A10). Assembly example: 10 µL 5 µM sur-siRNA + 10 µL 20–50 µM STD (PBS + 10% glycerol, on ice 30 min), then +10 µL 20 µM biotin-A10 (PBS pH 7.4, 30 min). |
| Formulation Format | A10-STD-(sur-siRNA) complex (aptamer-targeted CPP/DRBD fusion protein complex) |
| Formulation Components | STD fusion protein (streptavidin-3TAT-DRBD) + biotinylated PSMA aptamer A10 (2'-fluoro pyrimidine-modified) + SURVIVIN siRNA (FAM/Cy7-labeled for tracking in some assays). |
| Size Nm | |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | LNCaP (PSMA+ prostate cancer). Specificity controls: PC-3 (PSMA−), HeLa (PSMA−), HepG2 (PSMA−). |
| Animal Model | |
| Administration Route | |
| Output Type | Uptake/internalization (confocal microscopy + flow cytometry of FAM/Cy7-siRNA); functional effect (SURVIVIN mRNA by RT-qPCR at 48 h, protein by western blot at 72 h); apoptosis (Annexin V/PI flow cytometry ± cisplatin). |
| Output Value | |
| Output Units | |
| Output Notes | A10 provided PSMA-dependent binding to LNCaP (blocked by anti-PSMA). Complex delivered FAM-siRNA to cytoplasm with high % fluorescent cells (~99.8%), higher than A10-siRNA chimera (~39.9%). SURVIVIN knockdown and increased apoptosis vs controls were observed. |
| Toxicity Notes | Apoptosis increased after survivin knockdown; no separate general cytotoxicity panel reported beyond apoptosis assay context. |
| Curation Notes |