Sequence: Guanidinylated linear synthetic polypeptide gPADG19 (DP~19); poly(glutamate) derivative with near-quantitative side-chain guanidinylation
| Experiment Id | EXP001659 |
|---|---|
| Paper | Influences of chain length and conformation of guanidinylated linear synthetic polypeptides on nucle |
| Peptide | gPADG19 |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | yes |
| Label Confidence | high |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | |
| Peptide Concentration | Carrier stock 1.0 µg/µL; used to form nanocomplexes (NCs) at various N/P ratios |
| Rna Concentration | Typical: 50–100 nM in transfection; 30 nM for DLS/zeta; 100 nM for imaging; 50 nM siEF52 for TGS experiment |
| Mixing Ratio | N/P ratios tested: 1–80 (typical transfection N/P=10; imaging N/P=10; some controls N/P=80) |
| Formulation Format | Electrostatic polymer/siRNA nanocomplexes (NCs), incubated 30 min at 37°C |
| Formulation Components | GLSP (guanidinylated linear polypeptide) + siRNA in water/DEPC water (no additional formulation components) |
| Size Nm | 50.00 |
| Zeta Mv | 5.00 |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | HeLa (uptake/toxicity); HeLa-Luc (siGL4 luciferase knockdown); 293T (siEF52 TGS study; for gPALG100) |
| Animal Model | |
| Administration Route | |
| Output Type | in vitro functional RNA effect |
| Output Value | Luciferase knockdown up to ~80% under optimized conditions (depends on N/P, chain length, conformation); gPALG100/siEF52 reduced EF1A mRNA by ~77% and effect reversed by TSA+5-azaC (TGS evidence) |
| Output Units | |
| Output Notes | Silencing efficiency increased with N/P ratio and chain length; helical conformations (especially right-handed) higher efficiency. Confocal microscopy showed rapid nuclear localization of Cy3-siRNA (0.5–8 h). Endocytic inhibitor studies indicated clathrin (helical) vs caveolin (random coil); partial energy-independent membrane penetration for gPALGn. |
| Toxicity Notes | MTT cytotoxicity increased with chain length; GLSPs less toxic than PEI25k; NCs at N/P<10 maintained >80% viability (siRNA=100 nM). |
| Curation Notes |