scDynaBar: CRISPR-Cas9 dynamic barcoding enables single-cell temporal tracking using a molecular clock approach

Summary

This article presents scDynaBar, a novel method combining CRISPR-Cas9 dynamic barcoding with single-cell sequencing to track temporal cellular processes. The system uses genetic barcodes that accumulate mutations over time, which are sequenced alongside individual cell transcriptomes. The divergence of these barcodes from the original sequence serves as a molecular clock recording the timing of cellular events. The researchers demonstrate the method by tracking the transition from pluripotent to two-cell (2C)-like state in mouse embryonic stem cells, providing evidence for the transient nature of the 2C-like state. They also show consistent mutation rates across diverse cell types in a mouse gastruloid model, suggesting broad applicability for studying single-cell dynamics.