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

TY - JOUR

T1 - Efficient gene delivery to primary neuron cultures using a synthetic peptide vector system

AU - Collins, L

AU - Asuni, A A

AU - Anderton, B H

AU - Fabre, J W

PY - 2003/5/30

Y1 - 2003/5/30

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0037901399&partnerID=8YFLogxK

U2 - 10.1016/S0165-0270(03)00042-6

DO - 10.1016/S0165-0270(03)00042-6

M3 - Article

VL - 125

SP - 113

EP - 120

JO - Journal of Neuroscience Methods

JF - Journal of Neuroscience Methods

IS - 1-2

ER -