Nucleic Acids Research, 1992, Vol. 20, No. 4 763-770
© 1992
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Increased specificity for antisense oligodeoxynucleotide targeting of RNA cleavage by RNase H using chimeric methylphosphonodiester/phosphodiester structures
Department of Biochemistry, University of Liverpool PO Box 147, Liverpool, Merseyside, L69 3BX, UK
Received November 22, 1991. Revised January 20, 1992. Accepted January 20, 1992.
One of the inherent problems in the use of antisense oligodeoxynucleotides to ablate gene expression in cell cultures is that the stringency of hybridization in vivo is not subject to control and may be sub-optimal. Consequently, phosphodiester or phosphorothioate antisense effectors and non-targeted cellular RNA may form partial hybrids which are substrates for RNase H. Such processes could promote the sequence dependent Inappropriate effects recently reported in the literature. We have attempted to resolve this problem by using chimeric methylphosphonodiester/ phosphodiester oligodeoxynucleotides. In contrast to the extensive RNA degradation observed with all-phosphodiester oligodeoxynucleotides, highly modified chimeric antisense effectors displayed negligible, or undetectable, cleavage at non-target sites without significantly impaired activity at the target site. We also note that all of the all-phosphodiester oligodeoxynucleotides tested demonstrated inappropriate effects, and that such undesirable activity could vary widely between different sequences.
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