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EXP001699

Paper

Development of a peptide-siRNA nanocomplex targeting NF-κB for efficient cartilage delivery (2019)

Peptide

p5RHH

Sequence: VLTTGLPALISWIRRRHRRHC

RNA

siRNA

All experiment fields

Experiment Id EXP001699
Paper Development of a peptide-siRNA nanocomplex targeting NF-κB for efficient cartilage delivery
Peptide p5RHH
Delivery Success Class no
In Vivo Flag no
Uptake Confirmed yes
Label Confidence high
In Vitro Functional Effect yes
Endosomal Escape Evidence
Peptide Concentration For cell culture: 20 mM stock diluted 1:400 (~50 µM peptide) then mixed with siRNA; for explants: mixed to peptide:siRNA 1:10 (HBSS)
Rna Concentration 50 nM siRNA (cell culture); 500 nM siRNA (cartilage explants)
Mixing Ratio Peptide:siRNA 100:1 (cell culture); 1:10 (explant formulation in HBSS)
Formulation Format Self-assembling peptide/siRNA nanocomplex (NP)
Formulation Components p5RHH peptide + p65 siRNA smart pool (Dharmacon); self-assembled NP
Size Nm 55.00
Zeta Mv 12.00
Model Scope in_vitro
Model Type in vitro
Cell Lines Or Primary Cells HUVECs; human primary chondrocytes; human articular cartilage explants (IL-1β-stimulated OA cartilage)
Animal Model
Administration Route
Output Type in vitro functional knockdown / pathway suppression
Output Value Marked suppression of p65 expression in HUVECs (WB) and suppression of IL-1β-induced phospho-p65 activation in human cartilage explants persisting up to ~3 weeks after 48 h exposure.
Output Units
Output Notes NP prepared with peptide:siRNA ratio 100:1 for cell culture (50 nM siRNA final); for explants, peptide:siRNA ratio 1:10 in HBSS (500 nM siRNA final) incubated 10 min on ice; deep penetration of siRNA signal into cartilage up to ≥700 µm reported.
Toxicity Notes Cartilage surface viability unaffected by NP exposure (TUNEL, Fig. S1).
Curation Notes