Version 1
: Received: 10 July 2024 / Approved: 10 July 2024 / Online: 10 July 2024 (10:51:00 CEST)
How to cite:
Rai, M. S.; Sajid, M. I.; Moreno, J.; Parang, K.; Tiwari, R. K. Design, Synthesis, and Evaluation of Oleyl-WRH Peptides for siRNA Delivery. Preprints2024, 2024070833. https://doi.org/10.20944/preprints202407.0833.v1
Rai, M. S.; Sajid, M. I.; Moreno, J.; Parang, K.; Tiwari, R. K. Design, Synthesis, and Evaluation of Oleyl-WRH Peptides for siRNA Delivery. Preprints 2024, 2024070833. https://doi.org/10.20944/preprints202407.0833.v1
Rai, M. S.; Sajid, M. I.; Moreno, J.; Parang, K.; Tiwari, R. K. Design, Synthesis, and Evaluation of Oleyl-WRH Peptides for siRNA Delivery. Preprints2024, 2024070833. https://doi.org/10.20944/preprints202407.0833.v1
APA Style
Rai, M. S., Sajid, M. I., Moreno, J., Parang, K., & Tiwari, R. K. (2024). Design, Synthesis, and Evaluation of Oleyl-WRH Peptides for siRNA Delivery. Preprints. https://doi.org/10.20944/preprints202407.0833.v1
Chicago/Turabian Style
Rai, M. S., Keykavous Parang and Rakesh Kumar Tiwari. 2024 "Design, Synthesis, and Evaluation of Oleyl-WRH Peptides for siRNA Delivery" Preprints. https://doi.org/10.20944/preprints202407.0833.v1
Abstract
Delivering nucleic acid therapeutics across cell membranes is a significant challenge. Cell-penetrating peptides (CPPs) containing arginine (R), tryptophan (W), and histidine (H) show promise for siRNA delivery. To improve siRNA delivery and silencing a model STAT3 gene, we hypothesized that oleyl acylation to CPPs, specifically (WRH)n, would enhance STAT3 silencing efficiency in breast and ovarian cancer cells. Using Fmoc/tBu solid-phase peptide chemistry, we synthesized, purified, and characterized the oleyl-conjugated (WRH)n (n = 1-4) peptides. The peptide:siRNA complexes were non-cytotoxic at N/P 40 (~20 μM) against MDA-MB-231, MCF-7, SK-OV-3, and HEK-293 cells after 72 h incubation. All peptide:siRNA complexes showed serum stability at N/P ≥40. The synthesized conjugates, with a diameter of <100 nm, formed nano complexes with siRNA and exhibited stable range of zeta potential values (13-18 mV at N/P 40). Confocal microscopy and flow cytometry analysis provided qualitative and quantitative evidence of successful cellular internalization of siRNA. Peptides oleyl-(WRH)3 and oleyl-(WRH)4 showed ~60% and ~75% cellular uptake of siRNA, respectively, in both MDA-MB-231 and SK-OV-3 cells. Western blot analysis of oleyl-(WRH)4 demonstrated effective silencing of the STAT-3 gene, with ~75% silencing in MDA-MB-231 cells and ~45% in SK-OV-3 cells.
Keywords
acylation; arginine; breast cancer; cell-penetrating peptide; histidine; MDA-MB-231; oleic acid; RNA interference; siRNA; SK-OV-3; STAT-3; tryptophan; western blotting
Subject
Medicine and Pharmacology, Oncology and Oncogenics
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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