Exogenous expression of proteins in neurons using the biolistic particle delivery system.
Exogenous expression of genes in mammalian neurons represents a substantial experimental challenge because of the low efficiency of commercially available liposomal transfection reagents for nondividing cells and considerable toxicity of viral transfection systems. In this chapter, we discuss application of the "biolistic" particle delivery system for heterologous expression of genes in primary neuron cultures. The method is based on the direct introduction of cDNA of interest into the nucleus by penetration with DNA-coated gold particles. With this approach, cDNA expression is independent of cell cycling and proliferation and is similar to intranuclear microinjection, with both avoiding cDNA delivery through the cytosol. Examples of successful transfection using PDS of rat superior cervical ganglion and trigeminal ganglion neurons are discussed.
Gamper
Nikita
N
School of Biomedical Sciences, University of Leeds, UK.
Shapiro
Mark S
MS
D000818:Animals; D019470:Biolistics / Q000379:methods*; D002478:Cells, Cultured; D015870:Gene Expression; D008297:Male; D009474:Neurons / Q000378:metabolism*; D021381:Protein Transport; D011506:Proteins / Q000235:genetics*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017783:Superior Cervical Ganglion / Q000166:cytology; D014162:Transfection; D012668:Trigeminal Ganglion / Q000166:cytology; D010738:Type C Phospholipases / Q000378:metabolism
D011506:Proteins; D010738:Type C Phospholipases