Rational design and characterisation of a linear cell penetrating peptide for non-viral gene delivery

Emma M. McErlean, Monika Ziminska, Cian M. McCrudden, John W. McBride, Stephen P. Loughran, Grace Cole, Eoghan J. Mulholland, Vicky Kett, Niamh E. Buckley, Tracy Robson, Nicholas J. Dunne, Helen O. McCarthy*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

The design of a non-viral gene delivery system that can release a functional nucleic acid at the intracellular destination site is an exciting but also challenging proposition. The ideal gene delivery vector must be non-toxic, non-immunogenic, overcome extra- and intra-cellular barriers, protect the nucleic acid cargo from degradation with stability over a range of temperatures. A new 15 amino acid linear peptide termed CHAT was designed in this study with the goal of delivering DNA with high efficiency into cells in vitro and tissues in vivo. Rational design involved incorporation of key amino acids including arginine for nucleic acid complexation and cellular uptake, tryptophan to enhance hydrophobic interaction with cell membranes, histidine to facilitate endosomal escape and cysteine for stability and controlled cargo release. Six linear peptides were synthesised with strategic sequences and amino acid substitutions. Data demonstrated that all six peptides complexed pDNA to produce cationic nanoparticles less than 200 nm in diameter, but not all peptides resulted in successful transfection; indicating the influence of peptide design for endosomal escape. Peptide 4, now termed CHAT, was non-cytotoxic, traversed the plasma membrane of breast and prostate cancer cell lines, and elicited reporter-gene expression following intra-tumoural and intravenous delivery in vivo. CHAT presents an exciting new peptide for the delivery of nucleic acid therapeutics.

Original languageEnglish
Pages (from-to)1288
JournalJournal of Controlled Release
Volume330
Early online date21 Nov 2020
DOIs
Publication statusEarly online date - 21 Nov 2020

Bibliographical note

Funding Information:
Funding provided by the Department for the Economy, Northern Ireland

Publisher Copyright:
© 2020 Elsevier B.V.

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

Keywords

  • Cell penetrating
  • Gene delivery
  • Linear peptides
  • Non-viral
  • Nucleic acids

ASJC Scopus subject areas

  • Pharmaceutical Science

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