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Antisense RNA Inhibits Endogenous Gene Expression in Mouse Preimplantation Embryos: Lack of Double-Stranded RNA ``Melting'' Activity

Arturo Bevilacqua, Robert P. Erickson and Virginia Hieber
Proceedings of the National Academy of Sciences of the United States of America
Vol. 85, No. 3 (Feb. 1, 1988), pp. 831-835
Stable URL: http://www.jstor.org/stable/31346
Page Count: 5
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Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
Antisense RNA Inhibits Endogenous Gene Expression in Mouse Preimplantation Embryos: Lack of Double-Stranded RNA ``Melting'' Activity
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Abstract

β -Glucuronidase activity increases 60-fold from the 4-cell to the blastocyst stage during in vitro development of mouse preimplantation embryos, secondary to a 13-fold increase in β -glucuronidase mRNA. Injections of antisense RNA from a β -glucuronidase cDNA clone lacking the 5-untranslated region and the coding sequences for ≈ 150 N-terminal amino acids were effective in partially blocking the appearance of β -glucuronidase activity. Injection of the same RNA, capped with guanosine(5)triphospho(5$^{\prime}$)guanosine (GpppG), into each blastomere at the 4-cell stage yielded 75% inhibition of enzyme activity at the blastocyst stage. Injections of the sense strand or of an unrelated RNA did not alter the normal increase in activity of the enzyme. These results are in accord with our inability to detect RNA-duplex ``melting'' activity in 1-cell mouse embryos. We suggest that it may be possible to analyze genetics of mammalian development by antisense techniques.

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