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US 11,377,689 B2Granted
Chemical compositions and uses thereof
AssigneeNanoString Technologies, Inc.
InventorsJoseph M. Beechem, Dae Kim, Margaret Hoang, Mark Gregory, Erin Piazza, Denise Zhou
Life of this patent
Estimating the term…
Abstract
The present invention relates to, among other things, probes, compositions, methods, and kits for simultaneous, multiplexed detection and quantification of protein and/or nucleic acid expression in a user-defined region of a tissue, user-defined cell, and/or user-defined subcellular structure within a cell that are adaptable for use with existing sequencing technologies.
Claims30
A method for spatially detecting at least one target analyte in a first location and a second location of a tissue sample comprising:
- a) contacting the tissue sample with a plurality of nucleic acid probes, wherein each of the nucleic acid probes comprise a target binding domain that binds to the at least one target analyte, wherein the tissue sample [has been] is treated to facilitate binding of the nucleic acid probes to the target analyte;
- b) collecting the nucleic acid probes, or portions thereof, bound to the at least one target analyte in a first location of the tissue sample under conditions that release the nucleic acid probes, or portions thereof, from the first location of the tissue sample;
- c) collecting the nucleic acid probes, or portions thereof, bound to the at least one target analyte in a second location of the tissue sample under conditions that release the nucleic acid probes, or portions thereof, from the second location of the tissue sample;
- d) performing an extension reaction that incorporates at least one nucleic acid sequence that identifies the first location of the tissue sample into each of the nucleic acid probes, or portions thereof, collected in step (b), thereby forming a first plurality of extension products that comprise the nucleic acid probes, or portions thereof, collected in step (b) and the at least one nucleic acid sequence that identifies the first location of the tissue sample;
- e) performing an extension reaction that incorporates at least one nucleic acid sequence that identifies the second location of the tissue sample into each of the nucleic acid probes, or portions thereof, collected in step (c), thereby forming a second plurality extension products that comprise the nucleic acid probes, or portions thereof, collected in step (c) and the at least one nucleic acid sequence that identifies the second location of the tissue sample;
- and
- f) identifying the first plurality of extension products and the second plurality of extension products by sequencing the first plurality of extension products and the second plurality of extension products, thereby spatially detecting the at least one target analyte in the first location of the tissue sample and the second location of the tissue sample.
4 more elements
A method for spatially detecting at least one target analyte in a first location and a second location of a tissue sample comprising:
- a) contacting the tissue sample with a first plurality of nucleic acid probes and a second plurality of nucleic acid probes,
- wherein each of the nucleic acid probes in the first plurality of nucleic acid probes comprise a target binding domain that binds to the at least one target analyte,
- wherein each of the nucleic acid probes in the second plurality of nucleic acid probes comprise a target binding domain that binds to the at least one target analyte,
- wherein the tissue sample is treated to facilitate binding of the first plurality of nucleic acid probes and the second plurality of nucleic acid probes to the at least one target analyte;
- b) forming ligated probes by ligating each of nucleic acid probes from the first plurality of nucleic acid probes and each of nucleic acid probes from the second plurality of nucleic acid probes that are bound to each of identical molecules from the at least one target analyte;
- c) collecting the ligated probes, or portions thereof, bound to each of the identical molecules from the at least one target analyte in a first location of the tissue sample under conditions that release the ligated probes, or portions thereof, from the first location of the tissue sample;
- d) collecting the ligated probes, or portions thereof, bound to each of the identical molecules from the at least one target analyte in a second location of the tissue sample under conditions that release the probes, or portions thereof, from the second location of the tissue sample;
- e) performing an extension reaction that incorporates at least one nucleic acid sequence that identifies the first location of the tissue sample into each of the ligated probes, or portions thereof, collected in step (c), thereby forming a first plurality of extension products that comprise the ligated probes, or portions thereof, collected in step (c) and the at least one nucleic acid sequence that identifies the first location of the tissue sample;
- f) performing an extension reaction that incorporates at least one nucleic acid sequence that identifies the second location of the tissue sample into each of the ligated probes, or portions thereof, collected in step (d), thereby forming a second plurality extension products that comprise the ligated probes, or portions thereof, collected in step (d) and the at least one nucleic acid sequence that identifies the at least second location of the tissue sample;
- and
- g) identifying the first plurality of extension products and the second plurality of extension products by sequencing the first plurality of extension products and the second plurality of extension products, thereby spatially detecting the at least one target analyte in the first location of the tissue sample and the second location of the tissue sample.
8 more elements
Dependencies are inferred from the claim text.