Dual resonance energy transfer nucleic acid probes
First Claim
1. A composition for detection of a subject nucleic acid comprising,a. a first nucleic acid probe that hybridizes to a first nucleic acid target sequence on the subject nucleic acid, forms a stem-loop structure when not bound to the first nucleic acid target sequence, and incorporates a resonance energy transfer donor moiety;
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Accused Products
Abstract
Dual nucleic acid probes with resonance energy transfer moieties are provided. In particular, fluorescent or luminescent resonance energy transfer moieties are provided on hairpin stem-loop molecular beacon probes that hybridize sufficiently near each other on a subject nucleic acid, e.g. mRNA, to generate an observable interaction. The invention also provides lanthanide chelate luminescent resonance energy transfer moieties on linear and stem-loop probes that hybridize sufficiently near each other on a subject nucleic acid to generate an observable interaction. The invention thereby provides detectable signals for rapid, specific and sensitive hybridization determination in vivo. The probes are used in methods of detection of nucleic acid target hybridization for the identification and quantification of tissue and cell-specific gene expression levels, including response to external stimuli, such as drug candidates, and genetic variations associated with disease, such as cancer.
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Citations
50 Claims
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1. A composition for detection of a subject nucleic acid comprising,
a. a first nucleic acid probe that hybridizes to a first nucleic acid target sequence on the subject nucleic acid, forms a stem-loop structure when not bound to the first nucleic acid target sequence, and incorporates a resonance energy transfer donor moiety; - and
b. a second nucleic acid probe that hybridizes to a second nucleic acid target sequence on the subject nucleic acid, forms a stem-loop structure when not bound to the second nucleic acid target sequence, and incorporates a resonance energy transfer acceptor moiety, wherein the first nucleic acid target sequence and the second nucleic acid target sequence are separated by a number of nucleotides on the subject nucleic acid such that a resonance energy transfer signal from interaction between the donor moiety of the first nucleic acid probe and the acceptor moiety of the second nucleic acid probe can be detected to determine hybridization of both the first nucleic acid probe and the second nucleic acid probe to the subject nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 43, 44, 45, 46, 47, 48, 49, 50)
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24. A composition for detection of a subject nucleic acid comprising,
a. a first nucleic acid probe that hybridizes to a first nucleic acid target sequence on the subject nucleic acid, and incorporates a luminescence resonance energy transfer lanthanide chelate donor moiety; - and
b. a second nucleic acid probe that hybridizes to a second nucleic acid target sequence on the subject nucleic acid, and incorporates an organic resonance energy transfer acceptor moiety, wherein the first nucleic acid target sequence and the second nucleic acid target sequence are separated by a number of nucleotides on the subject nucleic acid such that a luminescence resonance energy transfer signal from interaction between the lanthanide chelate donor moiety of the first nucleic acid probe and the acceptor moiety of the second nucleic acid probe can be detected to determine hybridization of both the first nucleic acid probe and the second nucleic acid probe to the subject nucleic acid. - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42)
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Specification