In ovo transfection of chicken embryos using cationic liposomes.
It is reported that cationic liposomes are capable of transfecting embryos in unincubated fertile chicken eggs and that the cationic liposome, TransfectAce, has superior properties to Lipofectin. In order to determine the duration of expression of genes introduced in this way, embryos were transfected with an expression vector encoding the firefly luciferase cDNA under the control of the Rous sarcoma virus long terminal repeat (LTR). Luciferase activity could be observed consistently in day 3 embryos and activity was detectable up to day 8 of incubation. The relative expression of luciferase under the control of different viral promoters was compared in transfected chicken embryo fibroblasts and day 3 embryos. The cytomegalovirus immediate early promoter and the SV40 early promoter directed the highest amount of expression in fibroblasts while the Rous sarcoma virus LTR caused the highest amount of expression in embryos. Chicken embryo fibroblasts were transfected with the luciferase vector in order to examine duration of reporter gene expression in vitro. Luciferase expression was decreased exponentially over a 24-day period after which point luciferase activity could no longer be detected. These data suggest that stable integration of transfected DNA using liposomes is a rare event. Nevertheless, liposome-mediated transfection of embryos is suitable for the examination of promoter activity in vivo and may be a useful method to transfect genes to study embryonic development.
Rosenblum
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Department of Genetics and Molecular Biology, Merck Research Laboratories, Rahway, NJ 07065, USA.
Chen
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D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D002412:Cations; D002478:Cells, Cultured; D002642:Chick Embryo; D003587:Cytomegalovirus / Q000235:genetics; D004625:Embryo, Nonmammalian / Q000201:enzymology; D008081:Liposomes; D008156:Luciferases / Q000235:genetics; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection
D002412:Cations; D008081:Liposomes; D008156:Luciferases