Multiplexable tag-based reporter system
AssigneeNanoString Technologies, Inc.
InventorPhilippa J. WEBSTER
Life of this patent
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Abstract
The present invention relates to compositions and methods for the detection and quantification of individual target molecules in biomolecular samples. In particular, the invention relates to coded, labeled compositions comprising at least two probes hybridized to each other that are capable of binding to and identifying target molecules based on the probes' label codes. Methods of making and using such compositions are also provided. The compositions can be used in diagnostic, prognostic, quality control and screening applications.
Claims27
A composition comprising a first probe and a second probe,
- a. said first probe comprising
- i. a first region that comprising a first target-specific sequence;
- and
- ii. a second region that does not overlap with the first region and does not bind to the target molecule;
- b. said second probe comprising
- i. a first region that binds to the second region of said first probe;
- ii. a first label attachment region which is hybridized to a first RNA molecule, wherein the first RNA molecule is attached to one or more label monomers that emit light constituting a first signal;
- and
- iii. a second label attachment region, which is non-overlapping to the first label attachment region, and which is hybridized to a second RNA molecule, wherein the second RNA molecule is attached to one or more label monomers that emit light constituting a second signal, and wherein the label attachment regions do not overlap with the first region of the second probe.
A composition comprising a first probe and a second probe,
- a. said first probe comprising
- i. a first region comprising a first target-specific sequence;
- and
- ii. a second region that does not overlap with the first region and does not bind to the target molecule;
- b. said second probe comprising
- i. a first region that binds to the second region of said first probe;
- and
- ii. a second region that does not overlap with the first region and comprises at least one affinity moiety.
A composition pair comprising a first composition and a second composition, wherein the first composition comprises a first probe and a second probe,
- a. said first probe comprising
- i. a first region comprising a first target-specific sequence;
- and
- ii. a second region that does not overlap with the first region and does not bind to the target molecule;
- b. said second probe comprising
- i. a first region that binds to the second region of said first probe;
- and
- ii. a second region comprising at least one affinity moiety, wherein the first region does not overlap with the second region;
- and wherein the second composition comprises a third probe and a fourth probe,
- c. said third probe comprising
- i. a first region comprising a second target-specific sequence;
- and
- ii. a second region that does not bind to the target molecule, wherein the first region and the second region do not overlap;
- d. said fourth probe comprising
- i. a first region that binds to the second region of said third probe;
- ii. a first label attachment region which is hybridized to a first RNA molecule, wherein the first RNA molecule is attached to one or more label monomers that emit light constituting a first signal;
- and
- iii. a second label attachment region, which is non-overlapping to the first label attachment region, and which is hybridized to a second RNA molecule, wherein the second RNA molecule is attached to one or more label monomers that emit light constituting a second signal,
- wherein the first target-specific sequence and the second target-specific sequence bind to different regions of the same target molecule, and
- wherein the first and second probes of the first composition cannot bind to the third or fourth probe of the second composition, and
- wherein when said composition 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.
The composition of 9, wherein the second probe further comprises at least a first label attachment region which is hybridized to a first RNA molecule, wherein the first RNA molecule is attached to one or more label monomers that emit light constituting a first signal.
A composition comprising:
- a. a first region comprising a target-specific sequence;
- and
- b. a second non-overlapping region comprising any one of SEQ ID NOs:
- 1-1345, or a complement thereof.
Dependencies are inferred from the claim text.