Localized electroporation: a method for targeting expression of genes in avian embryos.
Avian embryos are a popular model for cell and developmental biologists. However, analysis of gene function in living embryos has been hampered by difficulties in targeting the expression of exogenous genes. We have developed a method for localized electroporation that overcomes some of the limitations of current techniques. We use a double-barreled suction electrode, backfilled with a solution containing a plasmid-encoding green fluorescent protein (GFP) and a neurophysiological stimulator to electroporate small populations of cells in living embryos. As many as 600 cells express GFP 24-48 h after electroporation. The number of cells that express GFP depends on the number of trains, the pulse frequency and the voltage. Surface epithelial cells and cells deep to the point of electroporation express GFP. No deformities result from electroporations, and neurons, neural crest, head mesenchyme, lens and otic placode cells have been transfected. This method overcomes some of the disadvantages of viral techniques, lipofection and in vivo electroporation. The method will be useful to biologists interested in tracing cell lineage or making genetic mosaic avian embryos.
Atkins
R L
RL
University of Victoria, BC, Canada.
Wang
D
D
Burke
R D
RD
D000818:Animals; D002452:Cell Count; D002642:Chick Embryo; D018274:Electroporation / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D008839:Microelectrodes; D018613:Microscopy, Confocal; D010957:Plasmids; D014162:Transfection
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins