Regenerative base editing enables deep lineage recording.

Area of Science:

• Molecular Biology
• Genetics
• Systems Biology

Background:

• Cell lineage tracing is crucial for understanding development and disease.
• Existing molecular recording systems face limitations in recording depth due to declining editing rates over time.
• Reconstructing cell lineage histories relies on heritable genetic barcodes.

Purpose of the Study:

• To develop a regenerative molecular recording system for deep lineage reconstruction.
• To overcome the limitations of exponentially declining editing rates in current systems.
• To enable high-density information storage and constant mutation accumulation.

Main Methods:

• Introduction of the hypercascade system, a regenerative molecular recording tool.
• Utilizing predictable A-to-G base editing to create new target sites progressively.
• Engineering a system with high density of editable target sites (4 per 20 bp).

Main Results:

• The hypercascade achieves an approximately constant rate of mutation accumulation.
• Demonstrated reconstruction of deep lineage relationships spanning 23 days and ~17 generations.
• Simulations indicate potential for parallel recording of chromatin state dynamics.

Conclusions:

• The hypercascade system significantly enhances the depth and accuracy of cell lineage reconstruction.
• This regenerative approach offers a flexible and broadly applicable tool for molecular recording.
• The technology holds promise for studying dynamic biological processes at an unprecedented resolution.