Design and Evaluation of a Stearylated Multicomponent peptide-siRNA Nanocomplex for Efficient Cellular siRNA Delivery

Abstract

Aim: To develop a new synthetic peptide-based nanoparticulate siRNA delivery system. Materials & methods: DEN-K(GALA)-TAT-K(STR) was generated by incorporating stearic acid into a multicomponent peptide (DEN-K(GALA)-TAT), containing a cationic poly-L-lysine dendron, an endosome-disrupting peptide GALA and a cell-penetrating peptide TAT(48–60). Its physicochemical characteristics, size, toxicity, cellular uptake and gene knockdown activity of the peptide/siRNA complexes were studied. Results: DEN-K(GALA)-TAT-K(STR) exhibited a pH-responsive behavior, which assists with endosomal escape. When siRNA was delivered by DEN-K(GALA)-TAT-K(STR), it showed a significantly enhanced cellular uptake, compared with the nonlipidic peptide. This system also displayed enhanced knockdown efficiency and reduced cytotoxicity over the widely used delivery system branched 25-kDa polyethyleneimine. Conclusion: Our stearylated multicomponent delivery system has great potential as an efficient siRNA delivery vector.

Keywords:

• cell-penetrating peptide
• endosome-disrupting peptide
• lipidic peptide
• nonviral vectors
• poly-L-lysine
• siRNA delivery

Supplementary data

To view the supplementary data that accompany this paper please visit the journal website at:www.tandfonline.com/doi/full/10.2217/epi-2016-0184

Acknowledgements

The authors wish to acknowledge B Gabrielli from Cell Cycle Group at the University of Queensland Diamantina Institute for providing a stable eGFP expressing HeLa cell line, and E Kaemmerer and A Peters (Calcium Signaling in Cancer Research Laboratory, School of Pharmacy, the University of Queensland) for assistance with the real-time PCR assay.

Financial & competing interests disclosure

This work was supported by an Australian Research Council (ARC) discovery project grant (DP130100952). Y Wan was supported by a University of Queensland International Postgraduate Research Scholarship (IPRS). PM Moyle was supported by an Australian National Health and Medical Research Council (NHMRC) postdoctoral training fellowship (569869). I Toth is funded by an ARC Australian Professorial Fellowship (DP110100212). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.