Oligonucleotide-based inhibition of embryonic gene expression.
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.
Driver
S E
SE
QIK Technology, Inc, Boston, MA, USA. sam.driver@qiktechnology.com
Robinson
G S
GS
Flanagan
J
J
Shen
W
W
Smith
L E
LE
Thomas
D W
DW
Roberts
P C
PC
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
D015820:Cadherins; D016228:Endothelial Growth Factors; D008222:Lymphokines; D016376:Oligonucleotides, Antisense; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors