Abstract
Artificial oligodeoxynucleotides (1,2), and later RNA (3) complementary to a particular gene, were shown to inhibit the expression of that gene in vertebrate cells. Transformation of plants with antisense transgenes was first used to block the expression of well-characterized plant genes in 1988 (4–6) and later it was found that sense transgenes would often similarly block endogenous gene expression (7–9). More recently, it has been shown that a single chimeric gene consisting of coding regions from two endogenous genes can silence the expression of both of the endogenous genes (10). This can be very useful if one of the two sequences is from a gene that gives rise to a color or other visual phenotype.
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References
Zamecnik, P. and Stephenson, M. (1978) Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. Proc. Natl. Acad. Sci. USA 75, 280–284.
Stephenson, M. and Zamecnik, P. (1978) Inhibition of Rous sarcoma viral RNA translation by a specific oligodeoxyribonucleotide. Proc.Natl. Acad. Sci. USA 75, 285–288.
Izant, J. G. and Weintraub, H. (1984) Inhibition of thymidine kinase gene expression by anti-sense RNA—a molecular approach to genetic analysis. Cell 36, 1007–1015.
van der Krol, A. R., Lenting, P. E., Veenstra, J., van der Meer, I. M., Koes, R. E., Gerats, A. G. M., Mol, J. N. M., and Stuitje, A. R. (1988) An anti-sense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature 333, 866–869.
Smith, C. J. S., Watson, C. F., Ray, J. Bird, C. R., Morris, P. C., Schuch, W., and Grierson, D. (1988) Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature 334, 724–726.
Sheehy, R. E., Kramer, M., and Hiatt, W. R. (1988) Reduction of polygalacturonase activity in tomato fruit by antisense RNA. Proc. Natl. Acad. Sci. USA 85, 8805–8809.
Napoli, C., Lemieux, C., and Jorgensen, R. (1990) Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. Plant Cell 2, 279–289.
van der Krol, A. R., Mur, L. A., Beld, M., Mol, J. N. M., and Stuitje, A. R. (1990) Flavonoid genes in petunia: addition of a limited number of gene copies may lead to a suppression of gene expression. Plant Cell 2, 291–299.
Smith, C. J. S., Watson, C. F., Bird, C. R., Ray, J., Schuch, W., and Grierson, D. (1990) Expression of a truncated tomato polygalacturonase gene inhibits expression of the endogenous gene in transgenic plants. Mol. Gen. Genet. 224, 477–481.
Seymour, G. B., Fray, R. G., Hill, P., and Tucker, G. A. (1993) Downregulation of two non-homologous endogenous tomato genes with a single chimaeric sense gene construct. Plant Mol. Biol. 23, 1–9.
Slater, A., Maunders, M. J., Edwards, K., Schuch, W., and Grierson, D. (1985) Isolation and characterisation of cDNA clones for tomato polygalacturonase and other ripening-related proteins. Plant Mol.Biol. 5, 137–147.
Smith, C. J. S., Slater, A., and Grierson, D. (1986) Rapid appearance of an mRNA correlated with ethylene synthesis encoding a protein of molecular weight 35 000. Planta 168, 94–100.
Holdsworth, M. J., Bird, C. J., Ray, J., Schuch, W., and Grierson, D. (1987) Structure and expression of an ethylene-related mRNA from tomato. Nucleic Acids Res. 15, 731–739.
Hamilton, A. J., Lycett, G. W., and Grierson, D. (1990) Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346, 284–287.
Ververidis, P. and John, P. (1991) Complete recovery in vitro of ethylene-forming enzyme activity. Phytochemistry 30, 725–727.
Hamilton, A. H., Bouzayen, M., and Grierson, D. (1991) Identification of a tomato gene for the ethylene-forming enzyme by expression in yeast. Proc. Natl. Acad. Sci. USA 88, 7434–7437.
John, I., Drake, R., Farrell, A., Cooper, W., Lee, P., Horton, P., and Grierson, D. (1995) Delayed leaf senescence in ethylene-deficient ACC-oxidase antisense tomato plants-molecular and physiological analysis. Plant J. 7, 483–490.
English, P. J., Lycett, G. W., Roberts, J. A., and Jackson, M. B. (1995) Increased 1-aminocyclopropane-1-carboxylic acid oxidase activity in shoots of flooded tomato plants raises ethylene production to physiologically active levels. Plant Physiol. 109, 1435–1440.
Cooper, W., Bouzayen, M., Hamilton, A., Barry, C., Rossall, S., and Grierson, D. (1998) Use of transgenic plants to study the role of ethylene and polygalacturonase during infection of tomato fruit by Colletotrichum gloeosporioides. Plant Pathol. 47, 308–316.
Pietrzk, M., Shillito, R. D., Hohn, T., and Potrykus, I. (1986) Expression in plants of two antibiotic resistance genes after protoplast transformation with a new plant expression vector. Nucleic Acids Res. 14, 5857–5868.
Bevan, M. W. (1984) Binary Agrobacterium vectors for plant transformation.Nucleic Acids Res. 12, 8711–8721.
Jones, C. G., Scothern, G. P., Lycett, G. W., and Tucker, G. A. (1998) Co-ordinated gene silencing by a single chimeric DNA construct.Planta 204, 499–505.
Bird, C. R., Ray, J. A., Fletcher, J. D., Boniwell, J. M., Bird, A. S., Teulieres, C., Blain, I., Bramley, P. M., and Schuch, W. (1991) Using antisense RNA to study gene function: inhibition of carotenoid biosynthesis in transgenic tomatoes. Biotechnology 9, 635–639.
Oeller, P. W. and Gutterson, N. (1997) A generic assay for gene silencing. 5th International Congress of Plant Molecular Biology, Singapore, September 1997, Abstract No 975.
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Lycett, G.W. (2000). The Identification of Ethene Biosynthetic Genes by Gene Silencing. In: Tucker, G.A., Roberts, J.A. (eds) Plant Hormone Protocols. Methods in Molecular Biology™, vol 141. Humana Press. https://doi.org/10.1385/1-59259-067-5:145
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DOI: https://doi.org/10.1385/1-59259-067-5:145
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