Gene transfer into mouse prepancreatic endoderm by whole embryo electroporation.
BACKGROUND
Understanding gene function in the developing pancreas is a major issue for pancreatic cell therapy. The in vivo analysis of gene function has essentially been performed by modulating gene expression in transgenesis. A faster and easier method is electroporation of mouse embryos. This technique, coupled with whole embryo culture, enables one to deliver genes and analyze their effects in a spatially and temporally regulated manner.


OBJECTIVE
We wanted to adapt the electroporation technique for gene transfer of whole e8.5 mouse embryos into the endoderm to allow expression of transgenes in the pancreas or liver.


RESULTS
Using two platinum plate electrodes, low voltage and a precise positioning of the embryo in the electroporation cuvette we could target and express DNA constructs in the prepancreatic or prehepatic territories, identified with cell markers. We also demonstrated that this technique is a valuable tool in the study of transcriptional regulation in the developing endoderm.


CONCLUSIONS
Targeted electroporation of whole embryos is a useful method of characterizing the gene network which controls pancreatic development.
Pierreux
Christophe E
CE
Hormone and Metabolic Research Unit, Institute of Cellular Pathology and Université Catholique de Louvain. Brussels, Belgium. christophe.pierreux@horm.ucl.ac.be
Poll
Aurélie V
AV
Jacquemin
Patrick
P
Lemaigre
Frédéric P
FP
Rousseau
Guy G
GG
D000818:Animals; D002463:Cell Membrane Permeability; D018274:Electroporation / Q000379:methods*; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000166:cytology / Q000201:enzymology / Q000502:physiology*; D047108:Embryonic Development; D004707:Endoderm / Q000737:chemistry / Q000201:enzymology / Q000502:physiology*; D005260:Female; D005455:Fluorescent Antibody Technique; D018507:Gene Expression Regulation, Developmental; D018014:Gene Transfer Techniques; D008099:Liver / Q000737:chemistry / Q000196:embryology / Q000502:physiology; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D051379:Mice; D008815:Mice, Inbred Strains; D010179:Pancreas / Q000737:chemistry / Q000196:embryology* / Q000502:physiology; D011247:Pregnancy; D014158:Transcription, Genetic
D008156:Luciferases