Nucleic Acids Research, Vol 26, Issue 10 2279-2285, Copyright © 1998 by Oxford University Press
J Ludwig, M Blaschke and BS Sproat
In this paper, we show that an adenosine to inosine mutation at position
15.1 changes the substrate specificity of the hammerhead ribozyme from
N16.2U16.1H17to N16.2C16.1H17(H represents A, C or U). This result extends
the hammerhead cleavage triplet definition from N16.2U16.1H17to the more
general N16.2Y16.1H17. Comparison of cleavage rates using I15.1ribozymes
for NCH triplets and standard A15.1 ribozymes for NUH triplets under single
turnover conditions shows similar or slightly enhanced levels of reactivity
for the I15. 1- containing structures. The effect of I15.1 substitution was
also tested in nuclease-resistant 2'- O -alkyl substituted derivatives
(oligozymes), showing a similar level of activity for the NUH and NCH
cleaving structures. The availability of NCH triplets that can be targeted
without loss of efficiency increases the flexibility of ribozyme targeting
strategies. This was demonstrated by an efficient cleavage of an HCV
transcript at a previously inaccessible GCA site in codon 2.
ARTICLES
Extending the cleavage rules for the hammerhead ribozyme: mutating adenosine15.1 to inosine15.1 changes the cleavage site specificity from N16.2U16.1H17 to N16.2C16.1H17
Innovir GmbH, Olenhuser Landstrasse 20b, D-37124 Rosdorf, Germany. jludwig@innovir.de
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