Targeted Activation of Cystic Fibrosis Transmembrane Conductance Regulator (2019)
HIV-1 Tat CPP (recombinant Tat peptide)
Sequence: GRKKRRQRRR
Gapmer antisense oligonucleotide (ASO; 20-mer)
| Experiment Id | EXP001998 |
|---|---|
| Paper | Targeted Activation of Cystic Fibrosis Transmembrane Conductance Regulator |
| Peptide | HIV-1 Tat CPP (recombinant Tat peptide) |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | no |
| Label Confidence | high |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | |
| Peptide Concentration | Tat CPP final concentration 200 mM (Tat:gapmer 200:10 mM) for electrostatic particles; incubated 2 h at 37°C before adding to cells; cells incubated 5 days before qPCR/ELISA readouts. |
| Rna Concentration | Gapmer final concentration 10 mM in Tat particle formulation (G10). |
| Mixing Ratio | Electrostatic particle: Tat CPP : gapmer final concentrations 200:10 mM (i.e., 20:1 molar ratio Tat:gapmer) after 2 h incubation at 37°C. |
| Formulation Format | Electrostatic CPP/ASO particles (Tat–gapmer complexes) |
| Formulation Components | Tat CPP (GRKKRRQRRR) complexed with BGas gapmer G10 (20-mer) via electrostatic binding. |
| Size Nm | |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | Human nasal epithelial cells (WT CFTR wt/wt; conditionally reprogrammed primary nasal cells) |
| Animal Model | |
| Administration Route | |
| Output Type | In vitro functional RNA effect: BGas knockdown and CFTR activation (qRT-PCR); CFTR protein by ELISA; functional iodide transport assay (EYFP quench) with/without VX809+VX770 |
| Output Value | CFTR mRNA increased (qRT-PCR) and CFTR protein increased (ELISA) after G10-Tat vs scramble; enhanced CFTR function in halide (iodide) transport assay (EYFP quench), including with VX809+VX770. |
| Output Units | |
| Output Notes | G10-Tat electrostatic particles target BGas lncRNA; functional readouts include CFTR transcript/protein increases and improved halide transport. |
| Toxicity Notes | IL-6 induction: no differential increase vs scramble; LDH cytotoxicity: modest increase vs control; overall described as relatively non-immunogenic/toxic under tested conditions. |
| Curation Notes |