Sequence: Stearyl-AGYLLGKLLOOLAAAALOOLL-NH2 (O = ornithine)
ASO (antisense oligonucleotide; steric-blocking)
| Experiment Id | EXP001728 |
|---|---|
| Paper | The nuclear concentration required for antisense oligonucleotide activity in myotonic dystrophy cell |
| Peptide | PepFect14 (PF14) |
| 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 | At highest ASO dose (1 µM), peptide-only control corresponds to 9 µM PF14; polyplex peptide concentration scales with ASO (9:1 molar). |
| Rna Concentration | ASO tested 0.1–1.0 µM for splice correction; imaging/uptake typically 0.1 µM Cy5-ASO. |
| Mixing Ratio | N/P = 3 (PF14), corresponding to peptide:ASO molar ratio 9:1; polyplexes stabilized ~1 h RT. |
| Formulation Format | Noncovalent CPP/ASO polyplex (charge-driven nanoparticle) |
| Formulation Components | PF14 + steric-blocking ASO (CAG5) in serum-free medium; noncovalent polyplexes. |
| Size Nm | |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | Immortalized human DM1 myoblasts (DM11 cl5) and unaffected myoblasts (C25); fibroblast validation in some experiments |
| Animal Model | |
| Administration Route | |
| Output Type | in vitro functional ASO effect (splice correction + MBNL1 foci dissolution) |
| Output Value | Dose-dependent correction of DM1-associated mis-splicing (MBNL1 exons 5/7, DMD exon 78) and improved myogenic capacity at low dose; nuclear ASO concentrations (IC50 ~340±200 nM) correlate with reduction of MBNL1 nuclear foci. |
| Output Units | |
| Output Notes | 24 h treatment + 24 h recovery for splice assays. Time-lapse confocal shows rapid uptake and homogeneous cytoplasmic+nuclear Cy5-ASO distribution in a subset of cells (25–45% nuclei positive). FLIM indicates intact ASO in nucleus; PF14 shields ASO from degradation and enables endosomal escape per LysoTracker-associated quenched fraction and proposed trafficking model. |
| Toxicity Notes | Higher PF14 polyplex doses (≥0.2–1 µM ASO) reduced viability and impaired differentiation; lowest dose (0.1 µM) improved differentiation morphology. |
| Curation Notes |