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EXP001435

Paper

Improving the efficacy of liposome-mediated vascular gene therapy via lipid surface modifications (2017)

Peptide

Octaarginine (R8)

Sequence: RRRRRRRR

RNA

siRNA

All experiment fields

Experiment Id EXP001435
Paper Improving the efficacy of liposome-mediated vascular gene therapy via lipid surface modifications
Peptide Octaarginine (R8)
Delivery Success Class no
In Vivo Flag no
Uptake Confirmed yes
Label Confidence high
In Vitro Functional Effect yes
Endosomal Escape Evidence
Peptide Concentration Surface density: 10 mol% STR-R8 of total lipids
Rna Concentration Gene silencing tested at 200–400 nM encapsulated siRNA (GAPDH or NC) in HASMCs; transfection 24 h serum-free then 24 h recovery
Mixing Ratio Transfection efficacy depended on lipid-to-siRNA w:w ratio (2.5:1–20:1) for 10 mol% formulation
Formulation Format R8-modified PEGylated neutral liposomes (R8-PLP)
Formulation Components Neutral PEGylated liposome (PLP) base: DOPC:cholesterol 7:3 mol + 10 mol% DSPE-PEG2000 + STR-R8 (10 mol%) + encapsulated siRNA; Ethanol injection with siRNA (50 µg) in 10 mM Tris-HCl pH 8.0; neutral formulations include 10 mM CaCl2 during assembly; dialyzed and extruded (100 nm)
Size Nm 40.20
Zeta Mv 10.50
Model Scope in_vitro
Model Type in vitro
Cell Lines Or Primary Cells Human aortic smooth muscle cells (HASMCs, primary; 49-year-old male donor)
Animal Model
Administration Route
Output Type In vitro functional effect: GAPDH mRNA knockdown by qPCR (normalized to 18S; compared to control or NC siRNA)
Output Value
Output Units
Output Notes Significant GAPDH silencing observed for R8-PLP 10 mol% at higher lipid:siRNA capacities (e.g., 400 nM GAPDH siRNA at 20:1 w:w ~41% relative expression; 200 nM at 10:1 w:w ~83% relative expression).
Toxicity Notes No significant cytotoxicity detected for R8-PLP 10 mol% vs untreated controls at 200 nM siRNA exposure.
Curation Notes