Widespread lipoplex-mediated gene transfer to vascular endothelial cells and hemangioblasts in the vertebrate embryo.
We report here a method that allows fast, efficient, and low-cost screening for gene function in the vascular system of the vertebrate embryo. Through intracardiac delivery of nucleic acids optimally compacted by a specific cationic lipid, we are able to induce in vivo endothelial cell-specific gain-of-function during development of the vascular network in the chick embryo. When the nucleic acids are delivered during the period of intraembryonic hematopoiesis, aortic hemangioblasts, the forerunners of the hematopoietic stem cells known to derive from the aortic endothelium, are also labeled. Similarly, we show that siRNA could be used to induce loss-of-function in vascular endothelial cells. This gene transfer technique was also applied to the mouse embryo with a high efficiency. The present method allows large-scale analysis and may represent a new and versatile tool for functional genomics.
Bollérot
Karine
K
UPMC, CNRS UMR7622, Laboratoire de Biologie du Développement, Bat C, 6ème étage, Case 24, 75252 Paris Cedex 05, France.
Sugiyama
Daisuke
D
Escriou
Virginie
V
Gautier
Rodolphe
R
Tozer
Samuel
S
Scherman
Daniel
D
Jaffredo
Thierry
T
D000818:Animals; D030801:Animals, Genetically Modified; D004622:Embryo, Mammalian / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism; D004730:Endothelium, Vascular / Q000196:embryology* / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D008081:Liposomes; D051379:Mice; D008822:Mice, Transgenic; D018919:Neovascularization, Physiologic / Q000235:genetics; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis / Q000235:genetics
D008081:Liposomes; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A