Methods and systems for analysis of cell lineage
The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing and analyte characterization from a single cell. Such polynucleotide processing may be useful for a variety of applications, including cell lineage analysis. Cell lineage analysis may comprise the use of one or more lineage tracing nucleic acid molecules. The disclosed methods may comprise using a lineage tracing nucleic acid molecule to identify a biological particle with one or more progenitor cells.
1. A method, comprising:
(a) partitioning a biological particle and a plurality of nucleic acid barcode molecules comprising a common barcode sequence into a partition, wherein said biological particle comprises an edited nucleic acid array and an analyte, wherein said edited nucleic acid array comprises an edited nucleic acid sequence that identifies an edited cell from which said biological particle was derived, and wherein said plurality of nucleic acid barcode molecules comprises (1) a first barcode molecule comprising said common barcode sequence and a first capture sequence configured to couple to said edited nucleic acid array; and (2) a second barcode molecule comprising said common barcode sequence and a second capture sequence configured to couple to said analyte or a second nucleic acid molecule corresponding to said analyte, wherein said first capture sequence or said second capture sequence lacks a poly(dT) sequence, wherein said biological particle is a cell, a cell nucleus, or a cell bead, and wherein said biological particle is not said edited cell, a nucleus of said edited cell, or cell bead comprising said edited cell or constituents thereof;
(b) coupling said first barcode molecule to said edited nucleic acid array to generate said first barcode molecule coupled to said edited nucleic acid array;
(c) coupling said second barcode molecule to said analyte or said second nucleic acid molecule corresponding to said analyte to generate said second barcode molecule coupled to (1) said analyte or (2) said second nucleic acid molecule corresponding to said analyte;
(d) synthesizing, using said first barcode molecule coupled to said edited nucleic acid array, a first nucleic acid product comprising said common barcode sequence and a sequence corresponding to said edited nucleic acid sequence; and
(e) synthesizing, using said second barcode molecule coupled to (1) said analyte or (2) said second nucleic acid molecule corresponding to said analyte, a second nucleic acid product comprising said common barcode sequence and (i) a sequence corresponding to said analyte or (ii) a sequence corresponding to said second nucleic acid molecule corresponding to said analyte.
2. The method of claim 1 , further comprising sequencing (i) said first nucleic acid product or a derivative thereof and (ii) said second nucleic acid product or a derivative thereof, to identify said common barcode sequence, said edited nucleic acid sequence, and said sequence of said second nucleic acid molecule corresponding to said analyte, wherein said common barcode sequence identifies said edited nucleic acid sequence and said analyte as having originated from said biological particle.
3. The method of claim 1 , wherein said common barcode sequence comprises identical barcode sequence segments.
4. The method of claim 3 , wherein said common barcode sequence of said first barcode molecule and said common barcode sequence of said second barcode molecule are identical.
5. The method of claim 1 , wherein said analyte is a ribonucleic acid (RNA) molecule.
6. The method of claim 5 , wherein said RNA molecule is a messenger RNA molecule (mRNA).
7. The method of claim 1 , wherein said nucleic acid molecule corresponding to said analyte is coupled to a labelling agent configured to couple to said analyte.
8. The method of claim 7 , wherein said second nucleic acid molecule corresponding to said analyte comprises an analyte barcode sequence that corresponds to said analyte.
9. The method of claim 7 , wherein said labelling agent is an antibody.
10. The method of claim 7 , wherein said analyte is a metabolite or a protein.
11. The method of claim 1 , wherein said plurality of nucleic acid barcode molecules is attached to a solid support.
12. The method of claim 11 , further comprising releasing said plurality of nucleic acid barcode molecules from said solid support.
13. The method of claim 11 , wherein said solid support is a bead.
14. The method of claim 1 , wherein (b) is performed in said partition.
15. The method of claim 1 , wherein (b) and (c) are performed in a partition.
16. The method of claim 1 , wherein said partition is an aqueous droplet in an emulsion.
17. The method of claim 1 , wherein said edited nucleic acid array nucleic acid molecule comprises an exogenous nucleic acid sequence.
18. The method of claim 1 , wherein prior to (a), said method comprises editing a nucleic acid molecule in an individual cell or group of cells to generate said edited cell.
19. The method of claim 1 , wherein said first capture sequence is different than said second capture sequence.
20. The method of claim 17 , wherein said exogenous nucleic acid sequence is a synthetic nucleic acid sequence.