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Nucleic Acids Research, 1993, Vol. 21, No. 24 5600-5603
© 1993

RNA

Activity of hammerhead ribozymes containing nonnucleotidic linkers

James B. Thomson, Thomas Tuschl and Fritz Eckstein*

Max-Planck-lnstitut fur Experimented Medizin, Hermann-Rein-StraGe 3 D-37075 Gottingen, Germany

*To whom correspondence should be addressed

Received September 20, 1993. Revised October 28, 1993. Accepted October 28, 1993.

Hammerhead ribozymes were synthesized in which the tetranucleotide loop II was replaced by non-nucleotidic linkers of 7, 13, 17 and 19 atoms length. Ribozymes with 17 and 19 atom linkers, in combination with a 4 base pair stem II, had catalytic efficiencies which were 2 fold increased to that of the parent ribozyme with a tetranucleotide loop. Ribozymes with these linkers, but in combination with a 2 base pair stem II, showed a 2 fold decrease in catalytic efficiency when compared to the parent ribozyme. Prolonged preincubation in the presence of MgCI2 was required for hexaethylene gylcol linker-modified ribozymes to obtain maximum activity and reproducible kinetic data.


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