Peptide/Cas9 nanostructures for ribonucleoprotein cell membrane transport and gene edition (2017)
PT24 (oleic aldehyde hydrazone amphiphile from peptide P)
Sequence: Arg/Leu-rich cationic peptide scaffold with two hydrazide (Hy) residues (exact sequence not in main text; schematic shows RRLKRRLLRRLKRL)
gRNA (CRISPR guide RNA) in Cas9 RNP
| Experiment Id | EXP001804 |
|---|---|
| Paper | Peptide/Cas9 nanostructures for ribonucleoprotein cell membrane transport and gene edition |
| Peptide | PT24 (oleic aldehyde hydrazone amphiphile from peptide P) |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | yes |
| Label Confidence | medium |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | yes |
| Peptide Concentration | PT24 10 mM (optimized); also tested 0.5–12.5 µM for gel retardation; uptake studies at 5 µM TmPT24 |
| Rna Concentration | Cas9 RNP 40–160 nM (dose-response); typical 80 nM or 160 nM; peptide:RNP ratio 62.5:1 (mol/mol) used in T7E1 assays |
| Mixing Ratio | RNP:PT24 = 1:62.5 (mol/mol) (editing assays) |
| Formulation Format | Non-covalent peptide amphiphile/Cas9 ribonucleoprotein nanoparticles |
| Formulation Components | PT24 amphiphilic CPP + SpCas9 protein complexed with gRNA (RNP); TAMRA-labelled PT24 (TmPT24) for uptake; DexPT24 for GR translocation assay |
| Size Nm | 270.00 |
| Zeta Mv | 4.20 |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | HeLa-EGFP |
| Animal Model | |
| Administration Route | |
| Output Type | Genome editing (EGFP knockout; % EGFP-negative cells by flow cytometry, 3 days) |
| Output Value | Screening shows EGFP-negative cells depending on aldehyde tail; PT24 (oleic aldehyde) best hit (typically >30% EGFP-negative in screen) |
| Output Units | |
| Output Notes | Hydrazone tail screening (PTn) identified T24 (oleic aldehyde) as best for EGFP editing; single incubation protocol. |
| Toxicity Notes | Lower toxicity vs Lipofectamine 2000 at comparable editing (MTT). |
| Curation Notes |