Direct Translocation as Major Cellular Uptake for CADY Self-Assembling Peptide-Based Nanoparticles (2011)
Sequence: Ac-GLWRALWRLLRSLWRLLWKA-cya
| Experiment Id | EXP001806 |
|---|---|
| Paper | Direct Translocation as Major Cellular Uptake for CADY Self-Assembling Peptide-Based Nanoparticles |
| Peptide | CADY |
| 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 | CADY 1.6–3.2 µM typical (e.g., 1.6 µM with 80 nM siRNA at 20:1; 3.2 µM with 80 nM siRNA at 40:1) |
| Rna Concentration | 80 nM siRNA (typical in uptake/knockdown assays); 20 nM used in Fura2 Ca2+ influx assay pulses |
| Mixing Ratio | CADY:siRNA = 20:1 to 40:1 (molar) |
| Formulation Format | Self-assembled non-covalent peptide/siRNA nanoparticles |
| Formulation Components | CADY peptide complexed with siRNA (FAM/Cy3/nanogold-labelled variants for tracking) |
| Size Nm | 100.00 |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | HeLa (primary mechanistic assays); CHO WT, CHO HS−/−, CHO gl−/− (GAG-deficient panels) for uptake comparisons |
| Animal Model | |
| Administration Route | |
| Output Type | Gene knockdown + uptake mechanism (Western blot; FACS; confocal; TEM; Ca2+ influx) |
| Output Value | High uptake (~60–90% FITC+ by FACS) and GAPDH protein reduction; activity retained with endocytosis inhibitors and at 4°C / with sodium azide |
| Output Units | |
| Output Notes | Minimal co-localization with transferrin/Rab5/caveolin; partial late lysotracker overlap. TEM shows many particles in cytosol without surrounding membrane, supporting direct translocation; FURA-2 shows transient Ca2+ influx indicating membrane permeabilization and resealing. |
| Toxicity Notes | No major increase in PI-positive cells during uptake assays at 37°C vs 4°C; viability largely maintained during treatments. |
| Curation Notes |