The use of Epstein-Barr virus-based shuttle vectors in rat PC12 cells.
1. The rat pheochromocytoma PC12 cell line has been a commonly used model for studies of neuronal development, function, and death. Thus the ability to transfect PC12 cells in an efficient manner and to manipulate their gene expression would enhance the usefulness of these cells. 2. We demonstrate that EBV-based vectors provide a useful expression system for gene manipulation in rat PC12 cells. 3. The EBV-based vectors replicate episomally in PC12 cells for at least 2 months, as evidence by their recovery from the transfected cells and by the digestion of the episomal plasmid with the isoschizomer MboI and DpnI restriction enzymes. 3. PC12 cells are efficiently transfected by EBV-based vectors both transiently and stably. 4. Transfection of PC12 cells with an EBV-based vector containing tau cDNA in the antisense orientation resulted in a decrease in the level of tau protein in the transfected cells. 5. The results demonstrate that EBV-based vectors can be a useful expression system for gene manipulation in PC12 cells.
Lindenboim
L
L
Department of Neurobiochemistry, George S. Wise Faculty of Life Sciences, Tel Aviv University, Israel.
Anderson
D
D
Stein
R
R
D000818:Animals; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000235:genetics*; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D016716:PC12 Cells / Q000187:drug effects; D010957:Plasmids; D051381:Rats; D014162:Transfection / Q000379:methods*; D016875:tau Proteins / Q000235:genetics / Q000378:metabolism
D016376:Oligonucleotides, Antisense; D016875:tau Proteins