High-throughput single-cell polyomics
Provided herein, in some embodiments, are devices, systems and methods for high-throughput single-cell polyomics (e.g., genomic, epigenomic, proteomic and/or phenotypic profile) analyses.
1. A polyomic multiplexing device, comprising:
a substrate comprising X rows intersecting (i) Y columns to form X*Y patches, and (ii) Z n+1 columns to form X*Z patches, wherein each of the X*Y patches comprises a unique nucleic acid barcode that is immobilized to the substrate and comprises a polyT sequence, wherein:
each of the rows comprises a different subset of barcoded nucleic acid strands of a first set of nucleic acid strands, and each of the Y columns comprises a different subset of barcoded nucleic acid strands of a second set of nucleic acid strands,
the nucleic acid strands of the first set are bound to nucleic acid strands of the second set to form the unique nucleic acid barcode at each of the X*Y patches,
the unique nucleic acid barcode comprises, in the 5′ to 3′ direction, a barcode sequence of the first set, a barcode sequence of the second set, and a polyT sequence,
each of the X*Z patches comprises an antibody immobilized to the substrate, and
n is zero or greater;
and
an array of microwells coupled to the substrate, wherein each microwell comprises one of the unique molecular barcodes immobilized the substrate and at least one of the antibodies immobilized to the substrate.
2. The polyomic multiplexing device of claim 1 , wherein X is at least 10.
3. The polyomic multiplexing device of claim 2 , wherein X is 10 to 20000.
4. The polyomic multiplexing device of claim 1 , wherein Y is at least 10.
5. The polyomic multiplexing device of claim 4 , wherein Y is 10 to 20000.
6. The polyomic multiplexing device of claim 1 , wherein n is at least 1, and each of the Z n+ 1 columns comprises a different antibody.
7. The polyomic multiplexing device of claim 6 , wherein n is at least 2, and each of the Z n+ 1 columns comprises a different antibody.
8. The polyomic multiplexing device of claim 1 , wherein the microwell array comprises at least 20 microwells.
9. The polyomic multiplexing device of claim 1 , wherein the nucleic acid strands of the first set of nucleic acid strands comprise, in the 5′ to 3′ direction: a promoter sequence, a sequencing adaptor sequence, a first barcode sequence and a first anchor sequence.
10. The polyomic multiplexing device of claim 1 , wherein the nucleic acid strands of the second set of nucleic acid strands comprise, in the 5′ to 3′ direction: a polyT sequence, a unique molecular identifier sequence, a second barcode sequence and a second anchor sequence, wherein the second anchor sequence is complementary to the first anchor sequence.
11. The polyomic multiplexing device of claim 1 , wherein the unique nucleic acid barcode comprises, in the 5′ to 3′ direction: a promoter sequence, a sequencing adaptor sequence, a first barcode sequence, a second barcode sequence, a unique molecular identifier, and a polyT sequence.
12. The polyomic multiplexing device of claim 1 , wherein the substrate comprises glass, silicon or silica.
13. The polyomic multiplexing device of claim 1 , wherein the substrate is coated with poly-1-lysine.
14. The polyomic multiplexing device of claim 1 , wherein each column and/or row has a width of 50-200 microns.
15. The polyomic multiplexing device of claim 1 , wherein each patch has an area of 400-40,000 μm 2 .
16. The polyomic multiplexing device of claim 1 , wherein patches within a single row and/or within a single column are separated from each other by 20-200 microns.
17. The polyomic multiplexing device of claim 1 , wherein patches between adjacent rows and/or between adjacent columns are separated from each other by 20-200 microns.