Efficient gene delivery to primary neuron cultures using a synthetic peptide vector system

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23 Citations (Scopus)

Abstract

A bi-functional, 31 amino acid synthetic peptide (polylysine-molossin) was evaluated for gene delivery to primary cultures of rat cerebral cortex neurons. Polylysine-molossin consists of an amino terminal domain of 16 lysines for electrostatic binding of DNA, and a 15 amino acid, integrin-binding domain at the carboxyl terminal. High levels of gene delivery were obtained with 20-30 muM chloroquine, with a synthetic fusogenic peptide at an optimal DNA: polylysine-molossin: fusogenic peptide w/w ratio of 1:3:0.2, and with the addition of low concentrations of Lipofectamine 2000 at an optimal DNA:polylysine-molossin:Lipofectamine 2000 w/w ratio of 1:3:0.5. With the best combination, > 30% of neurons strongly expressed the beta-galactosidase reporter gene, with no observable toxicity. DNA concentrations > 2 mug/ml were essential for efficient gene delivery. This synthetic peptide provides a safe, readily standardised and flexible DNA vector system well suited to ex vivo gene delivery to neurons for experimental and clinical applications. (C) 2003 Elsevier Science B.V. All rights reserved.
Original languageEnglish
Pages (from-to)113 - 120
Number of pages8
JournalJournal of Neuroscience Methods
Volume125
Issue number1-2
DOIs
Publication statusPublished - 30 May 2003

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