Gene delivery into primary cerebral cortical neurons by lentiviral vector.
Several studies have shown the ability of human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors to infect nondividing brain neurons. We are the first to show that primary embryonic cerebral cortical neurons can be efficiently transduced by an HIV-1-based lentiviral vector encoding enhanced green fluorescent protein (EGFP). We also describe the optimal conditions for the transduction of cerebral cortical neurons with lentiviral vectors, and the kinetic process of infection. The percentage of cells expressing EGFP is a function of the time in culture and virus dose. The highest percentage of EGFP-expression achieved was 46.77% at 4 days in vitro (DIV) with a multiplicity of infection (m.o.i.) of 20. The results show that lentiviral vectors are not only good prospects for in vivo gene delivery, but are also good candidates for in vitro studies of the function of gene products in primary cerebral cortical neurons.
Zhang
Yuzhi
Y
Department of Pediatrics, Peking University First Hospital, No. 8 Xi An Men Street, West District, Beijing 100034, PR China.
Wang
Hansen
H
Pan
Hong
H
Bao
Xinhua
X
Li
Meirong
M
Jin
Jing
J
Wu
Xiru
X
D000818:Animals; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D004622:Embryo, Mammalian / Q000166:cytology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009474:Neurons / Q000378:metabolism*; D051381:Rats; D014161:Transduction, Genetic; D016896:Treatment Outcome
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins