Peptide Nanoparticle Delivery of Charge-Neutral Splice-Switching Morpholino Oligonucleotides (2015)
Sequence: Stearoyl-(RXH)4 (X=Ahx)
PMO (phosphorodiamidate morpholino oligonucleotide; splice-switching)
| Experiment Id | EXP001016 |
|---|---|
| Paper | Peptide Nanoparticle Delivery of Charge-Neutral Splice-Switching Morpholino Oligonucleotides |
| Peptide | STR5 |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | no |
| Label Confidence | high |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | |
| Peptide Concentration | Formulated as nanoparticles; DMD model exon 23 skipping in differentiated H2K mdx myotubes (also tested in healthy H2K) |
| Rna Concentration | PMO 1 µM (25-mer DMD ON) |
| Mixing Ratio | peptide:PMO molar ratio 5:1 (screen); ratio series 2:1–7:1 explored for selected peptides |
| Formulation Format | noncovalent lipopeptide/PMO nanoparticles (co-incubation; complex formation) |
| Formulation Components | STR5 + PMO DMD (M23D; exon 23 skipping); 5'-GGCCAAACCTCGGCTTACCTGAAAT-3' |
| Size Nm | 133.00 |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | U2OS stable luciferase reporter (BTK intron XLA model); SMA type I fibroblasts GM03813; H2K mdx myotubes + healthy H2K myotubes |
| Animal Model | |
| Administration Route | |
| Output Type | in vitro splice switching (RT-PCR exon inclusion/skipping; luc reporter splice correction) |
| Output Value | Exon 23 skipping observed by nested RT-PCR; several novel lipopeptides outperformed PF14 in H2K myotubes (notably histidine-rich STR5). |
| Output Units | |
| Output Notes | Serum presence (20% FBS) showed similar exon skipping to serum-free transfection for selected formulations; peptide:PMO ratio affected activity depending on peptide cationicity. |
| Toxicity Notes | Myotube viability (MTS): no toxicity at working concentration (~5 µM peptide). St-RXR4 showed toxicity at 40 µM (without PMO). |
| Curation Notes |