NANOREPORTERS AND METHODS OF MANUFACTURING AND USE THEREOF
First Claim
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1. A probe pair comprising a first probe and a second probe,said first probe being a complex comprising:
- (a) a first molecule, comprising;
(i) a first label attachment region to which are attached one or more label monomers that emit light constituting a first signal;
(ii) a second label attachment region, which is non-overlapping with the first label attachment region, to which are attached one or more label monomers that emit light constituting a second signal; and
(b) a first target-specific sequence attached to the first molecule,said second probe being a second molecule, comprising (i) a second target-specific sequence;
(ii) optionally, a third label attachment region to which are attached one or more label monomers that emit light constituting a third signal; and
(iii) optionally, an affinity tag attached to said second molecule;
wherein the first target-specific sequence and the second target-specific sequence bind to different regions of the same target molecule,wherein the target molecule is a naturally occurring molecule or a cDNA of a naturally occurring molecule or the complement of said cDNA,and wherein when said probe pair is bound to its target molecule, the identity of the first and second signals and their locations relative to each other constitute at least part of a code that identifies the target molecule.
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Abstract
The present invention relates to compositions and methods for detection and quantification of individual target molecules in biomolecular samples. In particular, the invention relates to coded, labeled probes that are capable of binding to and identifying target molecules based on the probes'"'"' label codes. Methods of making and using such probes are also provided. The probes can be used in diagnostic, prognostic, quality control and screening applications.
120 Citations
47 Claims
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1. A probe pair comprising a first probe and a second probe,
said first probe being a complex comprising: -
(a) a first molecule, comprising;
(i) a first label attachment region to which are attached one or more label monomers that emit light constituting a first signal;
(ii) a second label attachment region, which is non-overlapping with the first label attachment region, to which are attached one or more label monomers that emit light constituting a second signal; and(b) a first target-specific sequence attached to the first molecule, said second probe being a second molecule, comprising (i) a second target-specific sequence;
(ii) optionally, a third label attachment region to which are attached one or more label monomers that emit light constituting a third signal; and
(iii) optionally, an affinity tag attached to said second molecule;wherein the first target-specific sequence and the second target-specific sequence bind to different regions of the same target molecule, wherein the target molecule is a naturally occurring molecule or a cDNA of a naturally occurring molecule or the complement of said cDNA, and wherein when said probe pair is bound to its target molecule, the identity of the first and second signals and their locations relative to each other constitute at least part of a code that identifies the target molecule. - View Dependent Claims (3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47)
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2. A probe pair comprising a first probe and a second probe,
said first probe being a complex comprising: -
(a) a first molecule, comprising;
(i) a first label attachment region to which are attached one or more label monomers that emit light constituting a first signal;
(ii) a second label attachment region to which are attached one or more label monomers that emit light constituting a second signal; and(b) a first target-specific sequence attached to the first molecule, said second probe being a second molecule, comprising (i) a second target-specific sequence;
(ii) optionally, a third label attachment region to which are attached one or more label monomers that emit light constituting a third signal; and
(iii) optionally, an affinity tag attached to said second molecule;wherein the first target-specific sequence and the second target-specific sequence bind to different regions of the same target molecule, wherein the first signal and the second signal are spatially or spectrally distinguishable; wherein the target molecule is a naturally occurring molecule or a cDNA of a naturally occurring molecule or the complement of said cDNA, and wherein when said probe pair is bound to its target molecule, the identity of the first and second signals and their locations relative to each other constitute at least part of a code that identifies the target molecule.
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Specification