Thiol-Cyanobenzothiazole Ligation for the Efficient Preparation of Peptide-PNA Conjugates (2019)
ApoE(140-150) CPP – thiol–maleimide linkage (C3)
Sequence: Ac-LRKLRKRLLR-Ahx-Cys-NH2
PNA (antisense oligonucleotide; 20-mer)
| Experiment Id | EXP000987 |
|---|---|
| Paper | Thiol-Cyanobenzothiazole Ligation for the Efficient Preparation of Peptide-PNA Conjugates |
| Peptide | ApoE(140-150) CPP – thiol–maleimide linkage (C3) |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | no |
| Label Confidence | high |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | |
| Peptide Concentration | Cell treatment: 5 µM conjugate (24 h; 4 h Opti-MEM + 20 h growth media). Stability: 100 µM conjugate in buffer + 10 mM GSH (37°C). |
| Rna Concentration | 5 µM (equimolar in conjugate). |
| Mixing Ratio | covalent 1:1 |
| Formulation Format | covalent CPP–PNA conjugate (1:1) |
| Formulation Components | ApoE(140-150) CPP (Ac-LRKLRKRLLR) covalently linked to 20-mer PNA targeting SMN2 exon 7 inclusion; thiosuccinimide linkage (thiol–maleimide) |
| Size Nm | |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | SMA patient-derived fibroblasts (Type I, GM03813) |
| Animal Model | |
| Administration Route | |
| Output Type | in vitro functional RNA effect (splice correction; RT-qPCR full-length SMN2 after pre-incubation in 10 mM GSH) |
| Output Value | Splice-correction decreased after GSH exposure: ~22% loss after 6 h and ~28% loss after 24 h vs untreated (-GSH). |
| Output Units | |
| Output Notes | LC-MS stability in 10 mM GSH: ~20% intact after 72 h. Maleimide linkage susceptible to thiol-exchange/cleavage under reducing conditions. |
| Toxicity Notes | Not reported (no dedicated cytotoxicity section for fibroblast treatment). |
| Curation Notes |