N-terminal modification of an LAH4-derived peptide increases mRNA delivery in the presence of serum (2024)
Sequence: KKALLAHALHLLAALALHLAHLLKKA
| Experiment Id | EXP000833 |
|---|---|
| Paper | N-terminal modification of an LAH4-derived peptide increases mRNA delivery in the presence of serum |
| Peptide | LAH4-A1-Sali |
| Delivery Success Class | no |
| In Vivo Flag | no |
| Uptake Confirmed | yes |
| Label Confidence | high |
| In Vitro Functional Effect | yes |
| Endosomal Escape Evidence | yes |
| Peptide Concentration | |
| Rna Concentration | |
| Mixing Ratio | Typically 5 µg peptide per 0.5 µg mRNA (HCT116) or 7 µg peptide per 1 µg mRNA (HEK293T GFP/Cy5 experiments); conditions varied. |
| Formulation Format | peptide/mRNA complexes (polyplex) |
| Formulation Components | Buffers used for complexation included 10 mM acetate pH5, 150 mM NaCl, or 10 mM phosphate buffer pH6.25 (also PBS, HBG in optimization). |
| Size Nm | 257.00 |
| Zeta Mv | |
| Model Scope | in_vitro |
| Model Type | in vitro |
| Cell Lines Or Primary Cells | HCT116; HEK293T |
| Animal Model | |
| Administration Route | |
| Output Type | Functional reporter expression (luciferase and/or GFP) |
| Output Value | |
| Output Units | |
| Output Notes | Modified LAH4-A1 peptides evaluated mainly in HCT116/HEK293T (luciferase/GFP); Tyr2 and Sali show improved activity in phosphate pH6.25 and improved serum tolerance. DLS in 10 mM phosphate pH6.25: 257 nm ±219 (PDI 0.218) with 10 µg peptide +0.8 µg FLuc mRNA. Uptake/association shown by flow cytometry with Cy5-labeled mRNA (all cells Cy5+). Endosomal acidification dependence shown: concanamycin A (H+-ATPase inhibitor) reduces transfection (suggesting acidification-dependent endosomal escape). |
| Toxicity Notes | Cell viability (HCT116) decreased in serum-free conditions; in presence of 10% serum, viability ~100% for Tyr2 and Sali; Sali more cytotoxic without serum. |
| Curation Notes |