Particle bombardment-mediated gene transfer and expression in rat brain tissues.
We have previously demonstrated that the particle bombardment method for gene transfer (Accell) provides a new means for transfection of various cell types in culture. In this study we evaluate its application to rat brain systems. Using a luciferase (luc) gene as a reporter, we obtained high levels of transient gene expression in primary cultures of fetal brain tissue. Reduced but significant levels were also detected in adult brain primary cultures. Both neuron and glial cells were transfected using this technique. The transient gene expression level obtained with Accell was at least 100-fold higher than that obtained with three other gene transfer methods. The relative strengths of four cellular and seven viral promoters were also evaluated in these cultures. In vivo gene expression was studied using freshly excised and bombarded fetal brain tissues which were immediately transplanted into caudate or intracortical brain tissues of adult host animals. Assays showed that luciferase activity was present in transplants for up to two months following gene transfer. In vitro and in vivo expression of a rat tyrosine hydroxylase (TH) gene, a candidate gene for treatment of Parkinson's disease, was also detected in this rat brain system. Our results suggest that the particle bombardment gene transfer technology can be employed as an effective method for ex vivo gene transfer into brain tissues.
Jiao
S
S
Department of Pediatrics and Medical Genetics, Waisman Center, University of Wisconsin, Madison 53705.
Cheng
L
L
Wolff
J A
JA
Yang
N S
NS
D000818:Animals; D001921:Brain / Q000196:embryology / Q000378:metabolism*; D002478:Cells, Cultured; D016332:Fetal Tissue Transplantation; D015870:Gene Expression; D006801:Humans; D008156:Luciferases / Q000235:genetics; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D051381:Rats; D014162:Transfection / Q000379:methods*; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
D008156:Luciferases; D014446:Tyrosine 3-Monooxygenase