Tissue-specific RNA interference in post-implantation mouse embryos using directional electroporation and whole embryo culture.
In mammals, embryonic development is more difficult to analyze than in non-mammalian species because this development occurs in utero. Interestingly, whole embryo culture allows the normal development of mouse post-implantation embryos for up to 2 days in vitro. One limitation of this technology has been the difficulty of performing loss-of-gene function studies in this system. RNA interference (RNAi), whereby double-stranded RNA molecules suppress the expression of complementary genes, has rapidly become a widely used tool for gene function analyses. We have combined the technologies of mouse whole embryo culture and RNAi to allow the molecular dissection of developmental processes. Here, we review the manipulation by topical injection followed by directional electroporation of endoribonuclease-prepared siRNA to demonstrate that this technology may be useful to knock down genes in a tissue- and region-specific manner in several organs of the developing mouse embryo.
Calegari
Federico
F
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307, Dresden, Germany.
Marzesco
Anne-Marie
AM
Kittler
Ralf
R
Buchholz
Frank
F
Huttner
Wieland B
WB
D000818:Animals; D018274:Electroporation; D046149:Embryo Culture Techniques / Q000295:instrumentation; D004622:Embryo, Mammalian / Q000033:anatomy & histology / Q000502:physiology*; D004722:Endoribonucleases / Q000378:metabolism; D051379:Mice; D009420:Nervous System / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism
D034741:RNA, Small Interfering; D004722:Endoribonucleases