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Updated: May 16, 2025
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
Published on: April 11, 2019
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BASELINE: A CRISPR Base Editing Platform for Mammalian-Scale Single-Cell Lineage Tracing.
Biorxiv : the Preprint Server for Biology
|April 1, 2025
Summary
We developed BASELINE, a CRISPR-based tool for high-resolution cell lineage tracing. It significantly increases recorded information and enables deep lineage reconstructions for developmental and disease studies.
Area of Science:
- Molecular Biology
- Genetics
- Developmental Biology
Background:
- Cell fate is determined by inherited lineage and environmental context.
- CRISPR-based lineage recording technologies face challenges in single-cell recovery, engineering complexity, and scale.
Purpose of the Study:
- To introduce BASELINE, a novel CRISPR-based system for high-resolution cell lineage tracing.
- To overcome limitations of existing lineage recording technologies.
Main Methods:
- Utilized CRISPR base editing with the Cas12a adenine base editor.
- Engineered cells with 50 synthetic target sites integrated into the genome.
- Applied BASELINE to a pancreatic cancer model for lineage recording and single-cell profiling.
Main Results:
- BASELINE records lineage-specific marks over biologically relevant intervals.
- Achieved over 4300 bits of information recording, a 50X increase over existing technologies.
- Enabled high-fidelity lineage reconstructions up to 40 cell divisions deep.
Conclusions:
- BASELINE offers a powerful tool for high-resolution lineage tracing in development and disease.
- The technology is suitable for complex, small, and distributed biological systems.
- Expects broad applicability in various lineage-tracing projects.
