Bi-specific D3 (both PMOs attached at Pip6a C-terminus; Dmd PMO click)
Sequence: RXRRBRRXRYQFLIRXRBRXRB
| Experiment Id | EXP001987 |
|---|---|
| Paper | Bi-specific splice-switching PMO oligonucleotides conjugated via a single peptide active in a mouse |
| Peptide | Bi-specific D3 (both PMOs attached at Pip6a C-terminus; Dmd PMO click) |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | no |
| Label Confidence | high |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | |
| Peptide Concentration | 0.5–1 µM conjugate (serum-free Opti-MEM, 4 h transfection; then DMEM/5% horse serum 20 h). |
| Rna Concentration | Same as conjugate concentration (PMO content equimolar to conjugate; for bi-specific constructs total PMO per conjugate contains two PMOs). |
| Mixing Ratio | Covalent bi-specific: one PMO amide-linked; the other click-conjugated at C-terminus (triazole) using Bpg alkyne with X/B spacers; in D3 the Dmd-targeting PMO is click-conjugated. |
| Formulation Format | Covalent CPP–PMO conjugate (P-PMO); bi-specific constructs carry two PMOs on one CPP via amide + triazole (click) or disulfide linkages. |
| Formulation Components | Pip6a CPP conjugated to PMO(s): Dmd exon23 PMO and/or Acvr2b exon5 PMO (see construct-specific linkage notes). |
| Size Nm | |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | H2K/mdx mouse myoblasts differentiated to myotubes (mdx dystrophin model) |
| Animal Model | |
| Administration Route | |
| Output Type | In vitro functional RNA effect: exon skipping (RT-PCR) for Dmd exon 23 and Acvr2b exon 5 |
| Output Value | High exon skipping for both targets; marginally more efficient than D2 in cells (RT-PCR). |
| Output Units | |
| Output Notes | Top-performing bi-specific design (with both PMOs at C-terminus). |
| Toxicity Notes | |
| Curation Notes |