A method is described for cleaving a nucleic acid substrate with a nucleic
acid enzyme at a cleavage site comprising mixing the substrate with the
enzyme, wherein the substrate includes a 7 nucleotide sequence with at
least 6 nucleotides 3' to the cleavage site and at least 1 nucleotide 5'
to the cleavage site and of formula: 5'-H'GNNHNN-3' wherein each N is a
nucleotide which may be the same or different, H is a nucleotide selected
from the group consisting of A, U, C, and T, and is the site of cleavage,
and H' is a ribonucleotide selected from the group consisting of A, U,
and C, wherein (i) the first nucleotide 3' to the cleavage site is
capable of forming a wobble pair with the enzyme, (ii) the second, third,
fifth, and sixth nucleotides 3' to the cleavage site are capable of
forming conventional Watson-Crick base pairs with the enzyme, (iii) the
fourth nucleotide 3' to the cleavage site is capable of forming a
non-conventional Watson-Crick base pair with the enzyme, and (iv) the
first nucleotide 5' to the cleavage site does not form a base pair with
the enzyme; and the enzyme comprises a substrate binding portion which is
capable of base pairing to the 6 nucleotides 3' to the cleavage site of
the substrate and which binding portion comprises the sequence:
3'-UNNXNN-5' wherein each N is a nucleotide which may be the same or
different, and X is a nucleotide selected from the group consisting of T,
U, A, and G, whereby binding of the substrate to the enzyme effects
cleavage of the substrate at the cleavage site.