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EXP001696

Paper

Oligoarginine-Bearing Tandem Repeat Penetration-Accelerating Sequence Delivers Protein to Cytosol via Caveolae-Mediated Endocytosis (2019)

Peptide

Pas2r12

Sequence: FFLIGFFLIG-rrrrrrrrrrrr-amide (Pas2 = FFLIG-FFLIG; r12 = D-dodeca-arginine, amidated)

RNA

All experiment fields

Experiment Id EXP001696
Paper Oligoarginine-Bearing Tandem Repeat Penetration-Accelerating Sequence Delivers Protein to Cytosol vi
Peptide Pas2r12
Delivery Success Class no
In Vivo Flag no
Uptake Confirmed yes
Label Confidence high
In Vitro Functional Effect
Endosomal Escape Evidence yes
Peptide Concentration Final to cells typically 20 µM CPP (after 45 min treatment; see Figure 2b/Methods).
Rna Concentration
Mixing Ratio Simple mixing with protein cargo (e.g., EGFP:CPP 15 µM : 20 µM final to cells).
Formulation Format Noncovalent CPP/protein complexes by simple mixing + incubation (no cross-linking required).
Formulation Components CPP + protein cargo (EGFP or IgG) in Opti-MEM; then DMEM added for incubation.
Size Nm
Zeta Mv
Model Scope in_vitro
Model Type in vitro
Cell Lines Or Primary Cells HEK293
Animal Model
Administration Route
Output Type protein cytosolic delivery (not RNA delivery)
Output Value Cytosolic/nuclear distribution of EGFP observed by CLSM (up to ~36% of cells with cytosolic EGFP after 16 h). Also delivers Alexa488-IgG to cytosol (~14–18% of cells at optimized conditions).
Output Units
Output Notes Mechanism: energy-dependent uptake requiring actin polymerization; inhibited strongly by caveolae-mediated endocytosis inhibitors (genistein, MβCD) and by caveolin-1 knockdown. Note: a siRNA against caveolin-1 is used as a mechanistic tool and is delivered using Lipofectamine RNAiMAX (not by Pas2r12).
Toxicity Notes MTT: mild viability decrease with increasing Pas2r12 concentration (e.g., ~85% at 20 µM; ~68% at 30 µM) in EGFP delivery condition.
Curation Notes