Creating maternal effect mutations in transgenic mice: antisense inhibition of an oocyte gene product.
Gene products present in mouse oocytes direct development until the two-cell stage and may be important in later development. Here, we demonstrate that expression of a specific maternal protein can be disrupted in mouse oocytes using transgenic antisense RNA technology. An oocyte-specific promoter (mZP3) was utilized to express antisense RNA directed against maternal mRNA encoding tissue-type plasminogen activator (tPA). Antisense expression results in reduced levels of tPA mRNA and enzyme activity in mouse oocytes. We also provide evidence for a novel mechanism of antisense-mediated translational inhibition, whereby the cytoplasmic polyadenylation of maternal tPA mRNA is altered. This strategy should prove applicable to functional studies of other murine maternal mRNAs in an in vivo environment.
Richards
W G
WG
Department of Pharmacology, State University of New York at Stony Brook 11794-8651.
Carroll
P M
PM
Kinloch
R A
RA
Wassarman
P M
PM
Strickland
S
S
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers; D015870:Gene Expression; D051379:Mice; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D009154:Mutation; D009865:Oocytes / Q000201:enzymology; D011061:Poly A / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D016372:RNA, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics*
D017931:DNA Primers; D011500:Protein Synthesis Inhibitors; D016372:RNA, Antisense; D012333:RNA, Messenger; D011061:Poly A; D010959:Tissue Plasminogen Activator