Abstract
Lanthanide complexes covalently attached to oligonucleotides have been shown to cleave RNA in a sequence-specific manner. Efficient cleavage, however, is at present limited to single-stranded RNA regions, as RNA in a duplex is considerably more resistant to strand scission. To overcome this limitation, we have designed and synthesised artificial nucleases comprising lanthanide complexes covalently linked to oligodeoxyribonucleotides which cleave a partially complementary RNA at a bulged site, in the duplex region. Strand scission occurs at or near the bulge. Cleavage of the RNA target by the metal complex can be addressed via the major or the minor groove. In an example of a competitive situation, where the cleavage moiety has access to both a bulge and a single-strand region, transesterification at the bulge is favoured. Such artificial ribonucleases may find application as antisense agents and as tools in molecular biology. In addition, the results may have importance for the design of artificial ribonucleases which are able to act with catalytic turnover.
Full Text
The Full Text of this article is available as a PDF (141.7 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brown R. S., Dewan J. C., Klug A. Crystallographic and biochemical investigation of the lead(II)-catalyzed hydrolysis of yeast phenylalanine tRNA. Biochemistry. 1985 Aug 27;24(18):4785–4801. doi: 10.1021/bi00339a012. [DOI] [PubMed] [Google Scholar]
- Hall J., Hüsken D., Pieles U., Moser H. E., Häner R. Efficient sequence-specific cleavage of RNA using novel europium complexes conjugated to oligonucleotides. Chem Biol. 1994 Nov;1(3):185–190. doi: 10.1016/1074-5521(94)90008-6. [DOI] [PubMed] [Google Scholar]
- Herschlag D., Cech T. R. Catalysis of RNA cleavage by the Tetrahymena thermophila ribozyme. 2. Kinetic description of the reaction of an RNA substrate that forms a mismatch at the active site. Biochemistry. 1990 Nov 6;29(44):10172–10180. doi: 10.1021/bi00496a004. [DOI] [PubMed] [Google Scholar]
- Hélène C., Toulmé J. J. Specific regulation of gene expression by antisense, sense and antigene nucleic acids. Biochim Biophys Acta. 1990 Jun 21;1049(2):99–125. doi: 10.1016/0167-4781(90)90031-v. [DOI] [PubMed] [Google Scholar]
- Kappen L. S., Goldberg I. H. Site-specific cleavage at a DNA bulge by neocarzinostatin chromophore via a novel mechanism. Biochemistry. 1993 Dec 7;32(48):13138–13145. doi: 10.1021/bi00211a024. [DOI] [PubMed] [Google Scholar]
- Milligan J. F., Matteucci M. D., Martin J. C. Current concepts in antisense drug design. J Med Chem. 1993 Jul 9;36(14):1923–1937. doi: 10.1021/jm00066a001. [DOI] [PubMed] [Google Scholar]
- Woolf T. M. To cleave or not to cleave: ribozymes and antisense. Antisense Res Dev. 1995 Fall;5(3):227–232. doi: 10.1089/ard.1995.5.227. [DOI] [PubMed] [Google Scholar]