Activatable probes and methods for in vivo gene detection
First Claim
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1. A probe set comprising:
- (a) a donor polymer comprising(i) a first polynucleotide binding domain complementary to a first region of a target polynucleotide flanked by first and second stem regions which hybridize in the absence of the target polynucleotide to form a stem-loop or random-coil structure, and(ii) a quantum dot; and
(b) an acceptor polymer comprising(i) a second polynucleotide binding domain complementary to a second region of the target polynucleotide flanked by first and second stem regions which hybridize in the absence of the target polynucleotide to form a stem-loop or random-coil structure, and(ii) at least one reporter;
wherein energy transfer occurs between the donor and the at least one reporter when the donor polymer and the acceptor polymer hybridize to the target polynucleotide and the quantum dot is exposed to an exciting amount of energy.
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Abstract
Probes for detecting a target polynucleotide are provided. One aspect provides a molecular beacon probe set wherein the donor molecular beacon comprises a quantum dot and an acceptor molecular beacon comprises at least one reporter. The probes optionally comprise a protein transduction domain, targeting signal, or a combination thereof. Methods for detecting target polynucleotides using the disclosed probes are also provided.
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Citations
20 Claims
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1. A probe set comprising:
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(a) a donor polymer comprising (i) a first polynucleotide binding domain complementary to a first region of a target polynucleotide flanked by first and second stem regions which hybridize in the absence of the target polynucleotide to form a stem-loop or random-coil structure, and (ii) a quantum dot; and (b) an acceptor polymer comprising (i) a second polynucleotide binding domain complementary to a second region of the target polynucleotide flanked by first and second stem regions which hybridize in the absence of the target polynucleotide to form a stem-loop or random-coil structure, and (ii) at least one reporter; wherein energy transfer occurs between the donor and the at least one reporter when the donor polymer and the acceptor polymer hybridize to the target polynucleotide and the quantum dot is exposed to an exciting amount of energy. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19)
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20. A method of detecting a target polynucleotide comprising:
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(a) delivering at least one probe pair to cell lysates, tissue extracts or the interior of a cell, wherein the probe pair comprises (i) a donor probe comprising a quantum dot and forms a stem-loop (or random coil) structure in the absence of a target polynucleotide and (ii) an acceptor probe comprising at least one reporter and forms a stem-loop (or random coil) structure in the absence of the target polynucleotide; and (b) exposing the quantum dot to an exciting amount of energy, wherein energy transfer occurs between the quantum dot and the reporter to produce a detectable signal when the donor probe and the acceptor probe hybridize to the target polynucleotide.
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Specification