Molecular torches
AssigneeGen-Probe Incorporated
InventorsMichael M. Becker, Gary P. Schroth
Life of this patent
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Abstract
The present invention features “molecular torches” and the use of molecular torches for detecting the presence of a target nucleic acid sequence. Molecular torches contain a target binding domain, a target closing domain, and a joining region. The target binding domain is biased towards the target sequence such that the target binding domain forms a more stable hybrid with the target sequence than with the target closing domain under the same hybridization conditions. The joining region facilitates the formation or maintenance of a closed torch.
Claims38
A molecular torch consisting essentially of:
- a target binding domain comprising nucleotide base recognition groups;
- a target closing domain comprising nucleotide base recognition groups, wherein said target binding domain is biased toward a target nucleic acid sequence, such that said target binding domain forms a more stable hybrid with said target sequence than with said target closing domain under strand displacement conditions;
- a joining region comprising one or more non-nucleotide linkers which joins said target binding domain and said target closing domain, wherein said joining region facilitates the formation of a target binding domain:target closing domain hybrid in the absence of said target sequence;
- and
- first and second labels, wherein said first label is attached to said target binding domain and said second label is attached to said target closing domain, and wherein said first and second labels interact to produce a first signal when said target binding domain and said target closing domain form a target binding domain:target closing domain hybrid and a second signal when said target binding domain and said target closing domain do not form said target binding domain:target closing domain hybrid, said first and second signals being distinguishable from each other.
A method for determining the presence of a target nucleic acid sequence in a sample, said method comprising the steps of:
- a) contacting said sample with a molecular torch consisting essentially of:
- a target binding domain comprising nucleotide base recognition groups;
- a target closing domain comprising nucleotide base recognition groups, wherein said target binding domain is biased toward said target sequence, such that said target binding domain forms a more stable hybrid with said target sequence than with said target closing domain under strand displacement conditions;
- a joining region comprising one or more non-nucleotide linkers which joins said target binding domain and said target closing domain, wherein said joining region facilitates the formation of a target binding domain:target closing domain hybrid in the absence of said target sequence;
- and
- first and second labels, wherein said first label is attached to said target binding domain and said second label is attached to said target closing domain, and wherein said first and second labels interact to produce a first signal when said target binding domain and said target closing domain form a target binding domain:target closing domain hybrid and a second signal when said target binding domain and said target closing domain do not form said target binding domain:target closing domain hybrid, said first and second signals being distinguishable from each other;
- b) exposing said sample to strand displacement conditions;
- and
- c) determining whether there has been a change in the interaction between said first and second labels after step b) as an indication of the presence or absence of a hybrid formed between said target binding domain and said target sequence in said sample.
Dependencies are inferred from the claim text.