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EXP001149

Paper

Advancing Antisense Oligonucleotide Delivery through Click Chemistry Based Chemical Conjugation with Designed Short Non-Cationic Peptides for Duchenne Muscular Dystrophy (2025)

Peptide

ETWWK

Sequence: ETWWK

RNA

ASO

All experiment fields

Experiment Id EXP001149
Paper Advancing Antisense Oligonucleotide Delivery through Click Chemistry Based Chemical Conjugation with
Peptide ETWWK
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 1.2 µM (functional); 5 µM used for uptake assays
Rna Concentration 1.2 µM (functional); 5 µM used for uptake assays
Mixing Ratio
Formulation Format covalent CPP–ASO conjugate (CuAAC click chemistry)
Formulation Components ETWWK peptide + exon51 ASO via triazole linker
Size Nm
Zeta Mv
Model Scope in_vitro
Model Type in vitro
Cell Lines Or Primary Cells C2C12; human DMD patient-derived myoblasts (KM1327, KM1328); human DMD patient-derived skin fibroblasts
Animal Model
Administration Route
Output Type dystrophin protein expression / splice correction
Output Value Increased dystrophin protein in KM1328 vs unconjugated ASO; Lipofectamine 2000 used as positive control
Output Units
Output Notes Uptake shown by fluorescence microscopy (C2C12) and flow cytometry (KM1328) using FAM-ETWWK-ASO vs FAM-ASO; uptake inhibited by chlorpromazine suggesting clathrin-mediated endocytosis. Lysosomal escape assessed by LysoTracker colocalization (reduced colocalization; nuclear shift). Functional readout: dystrophin upregulation by western blot after 24 h treatment at 1.2 µM.
Toxicity Notes MTT cytotoxicity evaluated in C2C12 and patient-derived myoblast/fibroblast cells; ETWWK-ASO reported as cytocompatible (arginine-containing variants noted as more cytotoxic).
Curation Notes