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

Efficient gene delivery to primary neuron cultures using a synthetic peptide vector system
Efficient gene delivery to primary neuron cultures using a synthetic peptide vector system
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 ?M 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 ?-galactosidase reporter gene, with no observable toxicity. DNA concentrations >2 ?g/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.
neuron, non-viral dna vector, peptide, gene therapy, rat
0165-0270
113-120
Collins, Louise
960297b1-03b1-490d-af01-08f38483966d
Asuni, Ayodeji A.
b1412b1b-9794-4705-aada-aed5d3da038f
Anderton, Brian H.
33184c75-df57-402f-ac03-abd14f51eb66
Fabre, John W.
262227d8-9c65-4150-8a88-65bd371fceac
Collins, Louise
960297b1-03b1-490d-af01-08f38483966d
Asuni, Ayodeji A.
b1412b1b-9794-4705-aada-aed5d3da038f
Anderton, Brian H.
33184c75-df57-402f-ac03-abd14f51eb66
Fabre, John W.
262227d8-9c65-4150-8a88-65bd371fceac

Collins, Louise, Asuni, Ayodeji A., Anderton, Brian H. and Fabre, John W. (2003) Efficient gene delivery to primary neuron cultures using a synthetic peptide vector system. Journal of Neuroscience Methods, 125 (1-2), 113-120. (doi:10.1016/S0165-0270(03)00042-6).

Record type: Article

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 ?M 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 ?-galactosidase reporter gene, with no observable toxicity. DNA concentrations >2 ?g/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.

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More information

Published date: May 2003
Keywords: neuron, non-viral dna vector, peptide, gene therapy, rat
Organisations: Centre for Biological Sciences

Identifiers

Local EPrints ID: 345530
URI: https://eprints.soton.ac.uk/id/eprint/345530
ISSN: 0165-0270
PURE UUID: ad1259cd-85e5-408b-a2e9-c31dfe27355d

Catalogue record

Date deposited: 12 Feb 2013 16:18
Last modified: 18 Jul 2017 05:09

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