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Nucleic Acids Research, 1992, Vol. 20, No. 17 4607-4612
© 1992

MOLECULAR BIOLOGY

Enhancement of ribozyme catalytic activity by a contiguous oligodeoxynucleotide (facilitator) and by 2'-O-methylation

John Goodchild*

Worcester Foundation for Experimental Biology 222 Maple Avenue, Shrewsbury, MA 01545, USA

* To whom correspondence should be addressed

Received April 14, 1992. Revised August 4, 1992. Accepted August 4, 1992.

RNA catalysts (ribozymes) designed to cleave sequences unique to viral RNA's might be developed as therapeutics. For this purpose, they would require high catalytic efficiency and resistance to nucleases. Reported here are two approaches that can be used in combination to improve these properties. First, catalytic efficiency can be improved by oligonucleo tides (facilitators) that bind to the substrate contiguously with the 3'-end of the ribozyme. Second, 2'-O-methylation of flanking sequences of the ribozyme increases catalytic activity as well as resistance to nucleases. In combination with a facilitator oligodeoxynucleotide, the cleavage rate was increased 20 fold over that of the unmodified ribozyme.


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