RECORDING AND MAPPING LINEAGE INFORMATION AND MOLECULAR EVENTS IN INDIVIDUAL CELLS
First Claim
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1. A method for characterizing lineage information or recording molecular events among cells in a cell population, comprising:
- introducing, over a time period of multiple cell cycle generations, a plurality of molecular changes in at least one of one or more genetic scratchpads in one or more cells in a cell population,wherein the cell population comprises cells that have developed for one or more cell cycle generations,wherein each genetic scratchpad in the one or more genetic scratchpads comprises a polynucleotide sequence and a plurality of target sites within the polynucleotide sequence, andwherein each of the plurality of molecular changes is associated with a target site among the plurality of target sites;
characterizing, at one or more time points during the time period, a status of molecular changes at each time point for the plurality of target sites in each genetic scratchpad in cells in the cell population, wherein the cells are essentially intact or undisrupted, wherein at least one time point in the one or more time points is two or more cell cycle generations from the beginning of the time period; and
establishing lineage connections or a sequence of molecular changes between cells from different cell cycle generations by comparing statuses of molecular changes of the cells, wherein the molecular changes may represent one or more molecular events.
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Abstract
Methods and systems for recording and mapping lineage information and molecular events in individual cells are provided. Molecular changes, which may result from random or specific molecular events, are introduced to defined regions in cells over multiple cell cycle generations. Techniques such as fluorescent imaging are applied to track and identify the molecular changes before such information is used for lineage analysis or for identifying key processes and key players in cellular pathways.
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Citations
25 Claims
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1. A method for characterizing lineage information or recording molecular events among cells in a cell population, comprising:
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introducing, over a time period of multiple cell cycle generations, a plurality of molecular changes in at least one of one or more genetic scratchpads in one or more cells in a cell population, wherein the cell population comprises cells that have developed for one or more cell cycle generations, wherein each genetic scratchpad in the one or more genetic scratchpads comprises a polynucleotide sequence and a plurality of target sites within the polynucleotide sequence, and wherein each of the plurality of molecular changes is associated with a target site among the plurality of target sites; characterizing, at one or more time points during the time period, a status of molecular changes at each time point for the plurality of target sites in each genetic scratchpad in cells in the cell population, wherein the cells are essentially intact or undisrupted, wherein at least one time point in the one or more time points is two or more cell cycle generations from the beginning of the time period; and establishing lineage connections or a sequence of molecular changes between cells from different cell cycle generations by comparing statuses of molecular changes of the cells, wherein the molecular changes may represent one or more molecular events. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24)
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25. A system for characterizing lineage information or molecular events among cells in a cell population, comprising:
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a housing component for one or more cells in a cell population, wherein a plurality of molecular changes is introduced over a time period of multiple cell cycle generations in at least one of one or more genetic scratchpads in one or more cells in a cell population, wherein the cell population comprises cells that have developed for one or more cell cycle generations, wherein each genetic scratchpad in the one or more genetic scratchpads comprises a polynucleotide sequence and a plurality of target sites within the polynucleotide sequence, and wherein each of the plurality of molecular changes is associated with a target site among the plurality of target sites; a characterization component, configured to characterize the cell population, at one or more time points during the time period, a status of molecular events at each time point for the plurality of target sites in each genetic scratchpad in cells in the cell population, wherein the cells are essentially intact or undisrupted, wherein at least one time point in the one or more time points is two or more cell cycle generations from the beginning of the time period; and an analytical component, designed to receive data from the characterization component and establish lineage connections or a sequence of molecular changes between cells from different cell cycle generations by comparing statuses of molecular changes of the cells, wherein the molecular changes may represent one or more molecular events.
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Specification