title: Oligonucleotide-based inhibition of embryonic gene expression.
abstract: We describe a technique to define gene function using antisense oligonucleotide (AS-ODN) inhibition of gene expression in mice. A single intravenous injection of an AS-ODN targeting vascular endothelial growth factor (VEGF) into pregnant mice between E7.5-8.5 resulted in a lack of primary angiogenesis. This enabled us to define the critical window required to inhibit VEGF expression and recapitulate the primary loss of function phenotype observed in VEGF (-/-) embryos. This phenotype was sequence-specific and time- and dose-dependent. Injection of an AS-ODN targeting a second gene, E-cadherin, into pregnant mice at E10 confirmed a hypothesized secondary phenotype. This is the first report of AS-ODN inhibition of gene expression in utero and provides a new strategy for target validation in functional genomics.
authors.affiliation: QIK Technology, Inc, Boston, MA, USA. sam.driver@qiktechnology.com
authors.forename: S E
authors.initials: SE
authors.lastname: Driver
authors.forename: G S
authors.initials: GS
authors.lastname: Robinson
authors.forename: J
authors.initials: J
authors.lastname: Flanagan
authors.forename: W
authors.initials: W
authors.lastname: Shen
authors.forename: L E
authors.initials: LE
authors.lastname: Smith
authors.forename: D W
authors.initials: DW
authors.lastname: Thomas
authors.forename: P C
authors.initials: PC
authors.lastname: Roberts
mesh_terms: D000818:Animals; D001483:Base Sequence; D015820:Cadherins / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D016228:Endothelial Growth Factors / Q000235:genetics*; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D008222:Lymphokines / Q000235:genetics*; D051379:Mice; D018345:Mice, Knockout; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D010641:Phenotype; D011247:Pregnancy; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
chemical_list: D015820:Cadherins; D016228:Endothelial Growth Factors; D008222:Lymphokines; D016376:Oligonucleotides, Antisense; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors