Synthetic recording and in situ readout of lineage information in single cells
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Summary
This summary is machine-generated.Scientists developed a new system called MEMOIR (memory by engineered mutagenesis with optical in situ readout) that records cellular lineage and events directly in the genome. This allows for in situ readout, reconstructing cell histories without direct imaging.
Area Of Science
- Cell biology
- Genetics
- Synthetic biology
Background
- Reconstructing cell lineage and dynamic event histories in native spatial contexts is challenging.
- Many biological processes occur in optically opaque or physically inaccessible environments, limiting direct imaging approaches.
Purpose Of The Study
- To develop a synthetic system for recording cellular lineage and event histories within the genome.
- To enable in situ readout of this recorded information from single cells.
Main Methods
- Developed MEMOIR (memory by engineered mutagenesis with optical in situ readout), a system using barcoded recording elements called scratchpads.
- Utilized CRISPR/Cas9 for targeted mutagenesis to alter scratchpad states irreversibly.
- Employed multiplexed single-molecule RNA fluorescence in situ hybridization (smFISH) for readout in single cells.
- Engineered mouse embryonic stem cells with MEMOIR components for proof-of-principle experiments.
Main Results
- Demonstrated progressive and stochastic alteration of scratchpads during cell proliferation.
- Successfully reconstructed lineage information from cell colonies by analyzing scratchpad states in situ.
- Inferred dynamic rates of gene expression state switching in embryonic stem cells by combining lineage reconstruction with endogenous gene expression analysis.
- Showcased through simulations the potential for parallel MEMOIR systems to record complex cellular event histories.
Conclusions
- MEMOIR provides a versatile platform for in situ, single-cell information recording and readout.
- This system overcomes limitations of direct imaging for studying cells in inaccessible contexts.
- Enables reconstruction of cell lineage and inference of dynamic cellular processes.