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Updated: Jan 24, 2026
13:36
A Practical Approach to Genetic Inducible Fate Mapping: A Visual Guide to Mark and Track Cells In Vivo
Published on: December 30, 2009
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Using Cre-recombinase-driven Polylox barcoding for in vivo fate mapping in mice
Weike Pei1, Xi Wang2,3, Jens Rössler2,3
1Division of Cellular Immunology, German Cancer Research Center, Heidelberg, Germany.
Nature Protocols
|May 22, 2019
Summary
We developed Polylox, a genetic barcoding system for high-resolution cell lineage tracing in mice. This method uses random DNA recombination to label individual stem cells, enabling detailed in vivo fate mapping across various tissues.
Area of Science:
- Developmental Biology
- Genetics
- Molecular Biology
Background:
- Fate mapping is crucial for understanding cell lineage and development in vivo.
- Current methods often lack the resolution to trace individual stem or progenitor cells.
- High diversity of genetic markers is needed for precise lineage tracing.
Purpose of the Study:
- To provide a detailed protocol for the Polylox genetic barcoding system.
- To enable high-resolution in vivo fate mapping in mice.
- To facilitate the study of cell lineages with individual cell resolution.
Main Methods:
- Utilizing the Polylox system for endogenous genetic barcoding in mice (Rosa26Polylox reporter).
- Inducing random DNA recombination via transient Cre recombinase activity.
- Employing single-molecule real-time (SMRT) DNA sequencing for barcode retrieval.
- Applying computational analysis for barcode identification and probability calculation.
Main Results:
- Polylox enables high-resolution fate mapping in various mouse tissues.
- The system allows for cell-type-specific and non-invasive labeling.
- High label diversity is achievable, surpassing some alternative methods.
- Experimental data generation takes ~2-3 weeks, with computational analysis under 2 days.
Conclusions:
- Polylox is a powerful tool for high-resolution in vivo fate mapping.
- It offers advantages in label diversity and non-invasive application.
- This protocol facilitates detailed studies of cell lineages in mice.
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