title: RNA inhibition of BMP-4 gene expression in postimplantation mouse embryos.
abstract: Short, hairpin RNA (shRNA) directed against bone morphogenetic protein 4 (Bmp-4) was delivered to early postimplantation staged mouse embryos via tail vein injection of pregnant dams. As early as 24 h postinjection, embryos expressed a DsRed marker and later exhibited defects of neural fold elevation and closure and of cardiac morphogenesis. Immunohistochemical analysis of sectioned embryos indicated that Bmp-4 protein was depleted and gene expression analysis indicated there was a reduction in Bmp-4 mRNA and an upregulation of the Bmp-4 antagonists, noggin and chordin, in embryos exposed to the shRNA, but not in control embryos. There was no change in the expression of Gata4, brachyury, or claudin6 in RNAi exposed embryos, indicating that RNA silencing was specific to Bmp-4 rather than producing widespread gene inhibition. Delivery of shRNA to embryos has the potential to specifically knockdown the expression of developmentally essential genes and to rescue gene mutations, significantly decreasing the time required to analyze the function(s) of individual genes in development.
mesh_terms: D000818:Animals; D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis* / Q000235:genetics; D057167:Claudins; D018076:DNA, Complementary / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism; D005326:Fetal Proteins / Q000378:metabolism; D005786:Gene Expression Regulation; D020868:Gene Silencing; D005822:Genetic Vectors; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000494:pharmacology; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D009154:Mutation; D009474:Neurons / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012313:RNA / Q000378:metabolism*; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020825:T-Box Domain Proteins / Q000378:metabolism; D013997:Time Factors; D015854:Up-Regulation; D000098908:Brachyury Protein
chemical_list: D055415:Bone Morphogenetic Protein 4; D019485:Bone Morphogenetic Proteins; D057167:Claudins; D018076:DNA, Complementary; D005326:Fetal Proteins; D008164:Luminescent Proteins; D008565:Membrane Proteins; D012313:RNA; D020825:T-Box Domain Proteins; D000098908:Brachyury Protein; C525810:Bmp4 protein, mouse; C413663:fluorescent protein 583; C122101:claudin 6