Abstract
beta-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 beta-glucuronidase mRNA. Injections of antisense RNA from a beta-glucuronidase cDNA clone lacking the 5'-untranslated region and the coding sequences for approximately equal to 150 N-terminal amino acids were effective in partially blocking the appearance of beta-glucuronidase activity. Injection of the same RNA, capped with guanosine(5')triphospho(5')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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- Bachvarova R., De Leon V. Polyadenylated RNA of mouse ova and loss of maternal RNA in early development. Dev Biol. 1980 Jan;74(1):1–8. doi: 10.1016/0012-1606(80)90048-2. [DOI] [PubMed] [Google Scholar]
- Bass B. L., Weintraub H. A developmentally regulated activity that unwinds RNA duplexes. Cell. 1987 Feb 27;48(4):607–613. doi: 10.1016/0092-8674(87)90239-x. [DOI] [PubMed] [Google Scholar]
- Chang L. J., Stoltzfus C. M. Gene expression from both intronless and intron-containing Rous sarcoma virus clones is specifically inhibited by anti-sense RNA. Mol Cell Biol. 1985 Sep;5(9):2341–2348. doi: 10.1128/mcb.5.9.2341. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
- Coleman J., Green P. J., Inouye M. The use of RNAs complementary to specific mRNAs to regulate the expression of individual bacterial genes. Cell. 1984 Jun;37(2):429–436. doi: 10.1016/0092-8674(84)90373-8. [DOI] [PubMed] [Google Scholar]
- Cox K. H., DeLeon D. V., Angerer L. M., Angerer R. C. Detection of mrnas in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Dev Biol. 1984 Feb;101(2):485–502. doi: 10.1016/0012-1606(84)90162-3. [DOI] [PubMed] [Google Scholar]
- Curci A., Bevilacqua A., Mangia F. Lack of heat-shock response in preovulatory mouse oocytes. Dev Biol. 1987 Sep;123(1):154–160. doi: 10.1016/0012-1606(87)90437-4. [DOI] [PubMed] [Google Scholar]
- Flach G., Johnson M. H., Braude P. R., Taylor R. A., Bolton V. N. The transition from maternal to embryonic control in the 2-cell mouse embryo. EMBO J. 1982;1(6):681–686. doi: 10.1002/j.1460-2075.1982.tb01230.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Green M. R., Maniatis T., Melton D. A. Human beta-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei. Cell. 1983 Mar;32(3):681–694. doi: 10.1016/0092-8674(83)90054-5. [DOI] [PubMed] [Google Scholar]
- Green P. J., Pines O., Inouye M. The role of antisense RNA in gene regulation. Annu Rev Biochem. 1986;55:569–597. doi: 10.1146/annurev.bi.55.070186.003033. [DOI] [PubMed] [Google Scholar]
- Harland R., Weintraub H. Translation of mRNA injected into Xenopus oocytes is specifically inhibited by antisense RNA. J Cell Biol. 1985 Sep;101(3):1094–1099. doi: 10.1083/jcb.101.3.1094. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hieber V. C. Cloning of a cDNA complementary to rat preputial gland beta-glucuronidase mRNA. Biochem Biophys Res Commun. 1982 Feb 26;104(4):1271–1278. doi: 10.1016/0006-291x(82)91387-0. [DOI] [PubMed] [Google Scholar]
- Holt J. T., Gopal T. V., Moulton A. D., Nienhuis A. W. Inducible production of c-fos antisense RNA inhibits 3T3 cell proliferation. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4794–4798. doi: 10.1073/pnas.83.13.4794. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Izant J. G., Weintraub H. Constitutive and conditional suppression of exogenous and endogenous genes by anti-sense RNA. Science. 1985 Jul 26;229(4711):345–352. doi: 10.1126/science.2990048. [DOI] [PubMed] [Google Scholar]
- Izant J. G., Weintraub H. Inhibition of thymidine kinase gene expression by anti-sense RNA: a molecular approach to genetic analysis. Cell. 1984 Apr;36(4):1007–1015. doi: 10.1016/0092-8674(84)90050-3. [DOI] [PubMed] [Google Scholar]
- Kim S. K., Wold B. J. Stable reduction of thymidine kinase activity in cells expressing high levels of anti-sense RNA. Cell. 1985 Aug;42(1):129–138. doi: 10.1016/s0092-8674(85)80108-2. [DOI] [PubMed] [Google Scholar]
- Liebhaber S. A., Cash F. E., Shakin S. H. Translationally associated helix-destabilizing activity in rabbit reticulocyte lysate. J Biol Chem. 1984 Dec 25;259(24):15597–15602. [PubMed] [Google Scholar]
- McGarry T. J., Lindquist S. Inhibition of heat shock protein synthesis by heat-inducible antisense RNA. Proc Natl Acad Sci U S A. 1986 Jan;83(2):399–403. doi: 10.1073/pnas.83.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melton D. A. Injected anti-sense RNAs specifically block messenger RNA translation in vivo. Proc Natl Acad Sci U S A. 1985 Jan;82(1):144–148. doi: 10.1073/pnas.82.1.144. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mizuno T., Chou M. Y., Inouye M. A unique mechanism regulating gene expression: translational inhibition by a complementary RNA transcript (micRNA). Proc Natl Acad Sci U S A. 1984 Apr;81(7):1966–1970. doi: 10.1073/pnas.81.7.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Newport J., Kirschner M. A major developmental transition in early Xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage. Cell. 1982 Oct;30(3):675–686. doi: 10.1016/0092-8674(82)90272-0. [DOI] [PubMed] [Google Scholar]
- Newport J., Kirschner M. A major developmental transition in early Xenopus embryos: II. Control of the onset of transcription. Cell. 1982 Oct;30(3):687–696. doi: 10.1016/0092-8674(82)90273-2. [DOI] [PubMed] [Google Scholar]
- Nielsen D. A., Shapiro D. J. Preparation of capped RNA transcripts using T7 RNA polymerase. Nucleic Acids Res. 1986 Jul 25;14(14):5936–5936. doi: 10.1093/nar/14.14.5936. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oshima A., Kyle J. W., Miller R. D., Hoffmann J. W., Powell P. P., Grubb J. H., Sly W. S., Tropak M., Guise K. S., Gravel R. A. Cloning, sequencing, and expression of cDNA for human beta-glucuronidase. Proc Natl Acad Sci U S A. 1987 Feb;84(3):685–689. doi: 10.1073/pnas.84.3.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rebagliati M. R., Melton D. A. Antisense RNA injections in fertilized frog eggs reveal an RNA duplex unwinding activity. Cell. 1987 Feb 27;48(4):599–605. doi: 10.1016/0092-8674(87)90238-8. [DOI] [PubMed] [Google Scholar]
- Rosenberg U. B., Preiss A., Seifert E., Jäckle H., Knipple D. C. Production of phenocopies by Krüppel antisense RNA injection into Drosophila embryos. Nature. 1985 Feb 21;313(6004):703–706. doi: 10.1038/313703a0. [DOI] [PubMed] [Google Scholar]
- Rubenstein J. L., Nicolas J. F., Jacob F. L'ARN non sens (nsARN): un outil pour inactiver spécifiquement l'expression d'un gène donné in vivo. C R Acad Sci III. 1984;299(8):271–274. [PubMed] [Google Scholar]
- Watson G., Felder M., Rabinow L., Moore K., Labarca C., Tietze C., Vander Molen G., Bracey L., Brabant M., Cai J. D. Properties of rat and mouse beta-glucuronidase mRNA and cDNA, including evidence for sequence polymorphism and genetic regulation of mRNA levels. Gene. 1985;36(1-2):15–25. doi: 10.1016/0378-1119(85)90065-4. [DOI] [PubMed] [Google Scholar]
- Whitten W. K., Biggers J. D. Complete development in vitro of the pre-implantation stages of the mouse in a simple chemically defined medium. J Reprod Fertil. 1968 Nov;17(2):399–401. doi: 10.1530/jrf.0.0170399. [DOI] [PubMed] [Google Scholar]
- Wormington W. M. Stable repression of ribosomal protein L1 synthesis in Xenopus oocytes by microinjection of antisense RNA. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8639–8643. doi: 10.1073/pnas.83.22.8639. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wudl L., Chapman V. The expression of beta-glucuronidase during preimplantation development of mouse embryos. Dev Biol. 1976 Jan;48(1):104–109. doi: 10.1016/0012-1606(76)90049-x. [DOI] [PubMed] [Google Scholar]
- Yatziv S., Weiss L., Morecki S., Fuks Z., Slavin S. Long-term enzyme replacement therapy in beta-glucuronidase--deficient mice by allogeneic bone marrow transplantation. J Lab Clin Med. 1982 Jun;99(6):792–797. [PubMed] [Google Scholar]