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EXP001459

Paper

Cyclic peptide-based nanostructures as efficient siRNA carriers (2018)

Peptide

CP III

Sequence: Cyclo[WWGGGRRRRGGG]

RNA

siRNA

All experiment fields

Experiment Id EXP001459
Paper Cyclic peptide-based nanostructures as efficient siRNA carriers
Peptide CP III
Delivery Success Class no
In Vivo Flag no
Uptake Confirmed yes
Label Confidence high
In Vitro Functional Effect yes
Endosomal Escape Evidence
Peptide Concentration Uptake: 30 µM (FACS) or ~8 µM (confocal). Knockdown: 24 µM.
Rna Concentration Uptake: 200 nM (FACS) or ~80 nM (confocal). Knockdown: 600 nM.
Mixing Ratio Confocal: 0.04 nmol FAM-siRNA + 4 nmol peptide in 500 µL DMEM, 30 min complexation. FACS: 200 nM FAM-siRNA + 30 µM peptide in Opti-MEM, 30 min. Western blot: 24 µM peptide + 600 nM GAPDH siRNA in 500 µL serum-free DMEM, 30 min; incubate cells 3 h then add complete media and culture 48 h.
Formulation Format Self-assembled cyclic peptide nanostructures / peptide–siRNA nanocomplexes (noncovalent)
Formulation Components CP III + GAPDH siRNA (FAM-labeled for uptake; unlabeled for knockdown)
Size Nm 238.50
Zeta Mv
Model Scope in_vitro
Model Type in vitro
Cell Lines Or Primary Cells HCT116 human colon cancer cells
Animal Model
Administration Route
Output Type Uptake (confocal microscopy + flow cytometry) and functional gene silencing (GAPDH protein by Western blot)
Output Value 2.4
Output Units
Output Notes Moderate uptake by confocal microscopy and FACS (55.4% FAM-siRNA positive vs 2.3% siRNA alone). Significant GAPDH protein knockdown (~2.4-fold vs control), comparable to lipofectamine.
Toxicity Notes Cyclic peptides showed >90% viability at 10 µM (24 h). siRNA/peptide complexes showed no significant cytotoxicity except CP I showed some cytotoxicity relative to other complexes; lipofectamine 2000 showed significant cytotoxicity.
Curation Notes