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Nucleic Acids Research, 2000, Vol. 28, No. 9 1859-1863
© 2000 Oxford University Press

Flexible non-nucleotide linkers as loop replacements in short double helical RNAs

Werner Pils1 and Ronald Micura1,2,*

1Institut für Chemie, Johannes Kepler Universität, Altenbergerstrasse 69, A-4040 Linz, Austria and 2Laboratorium für Organische Chemie, ETHZ, Universitätsstrasse 16, CH-8092 Zürich, Switzerland

Ethylene glycol oligomers have been studied systematically as non-nucleotide loop replacements in short hairpin oligoribonucleotides. Structural optimization concerns the length of the linkers and is based on the thermodynamic stabilities of the corresponding duplexes. The optimum linker is derived from heptakis (ethylene glycol) provided that the duplex end to be bridged comprises solely the terminal base pair; the optimum linker is derived from hexakis(ethylene glycol) if a dangling unpaired nucleotide is incorporated into the loop. Moreover, these linkers have been compared to other commonly used linker types which consist of repeating units of tris- or tetrakis(ethylene glycol) phosphate, or of 3-hydroxypropane-1-phosphate. In all cases, the correlation between linker length and duplex stability is independent of the kind of counter ions used (Na+, Na+/Mg2+, K+ or Li+). Furthermore, all duplexes with non-nucleotide loop replacements are less stable than those with the corresponding standard nucleotide loop. The results corroborate that the linkers are solvent-exposed and do not specifically interfere with the terminal nucleotides at the bridged duplex end.

* To whom correspondence should be addressed at: Institut für Chemie, Johannes Kepler Universität, Altenbergerstrasse 69, A-4040 Linz, Austria. Tel: +43 732 2468 780; Fax: +43 732 2468 747; Email: micura@gmx.net


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