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US 2021/0172934 A1Published application
Single cell cloning approaches for biological studies
OwnerSandra S. McAllister, Jessica F. Oliveinventors
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Abstract
Clonally derived barcoded cell populations and methods of generating the same, as well as methods of use thereof, e.g., to evaluate heterogeneity of a starting population of cells.
Claims22
A method of creating a plurality of barcoded clonal populations (BCPs), the method comprising:
- (a) providing an initial heterogeneous population of cells;
- (b) dividing the initial heterogeneous population of cells into separate cultures, each culture comprising a single cell from the initial heterogeneous population;
- (c) maintaining the single cells in culture to provide a plurality of stable single cell-derived monoclonal populations;
- and
- (d) introducing individual identifying nucleic acid sequences into each cell of the plurality of stable single cell-derived monoclonal populations;
- to thereby create a plurality of barcoded clonal populations (BCPs).
3 more elements
A method of selecting a therapy for a subject who has cancer, the method comprising:
- creating a plurality of barcoded clonal populations (BCPs) by a method comprising:
- (a) providing an initial heterogeneous population of cells from the cancer in the subject;
- (b) dividing the initial heterogeneous population of cells into separate cultures, each culture comprising a single cell from the initial heterogeneous population;
- (c) maintaining the single cells in culture to provide a plurality of stable single cell-derived monoclonal populations;
- and
- (d) introducing individual identifying nucleic acid sequences into each cell of the plurality of stable single cell-derived monoclonal populations;
- to thereby create a plurality of barcoded clonal populations (BCPs);
- (e) mixing equal numbers of each BCP to create a barcoded polyclonal population of cells (BPP);
- (f) exposing the BPP to a candidate therapeutic compound;
- and
- determining one or both of identity and relative abundance of each BCP in the BPP.
8 more elements
Dependencies are inferred from the claim text.