The genetic and functional heterogeneity of tumors imposes the challenge of understanding how a cancer progresses, evolves and adapts to treatment at the subclonal level. Therefore, there is a critical need for methods that enable profiling of individual cancer cell lineages. Here, we report a novel system that couples an established DNA barcoding technique for lineage tracing with a controlled DNA barcode-guided lineage isolation (B-GLI). B-GLI allows both high-complexity of lineage tracing and effective isolation of individual clones by CRISPRa-mediated induction of puromycin resistance, making it possible to unbiasedly trace, isolate, and study individual cancer cell lineages. We present an experimental evaluation of the system performance in isolation of lineages and outline a comprehensive workflow for B-GLI applications. We believe the system has broad applications aimed at molecular and phenotypic profiling of individual lineages in heterogeneous cell populations.

DNA barcode-guided lentiviral CRISPRa tool to trace and isolate individual clonal lineages in heterogeneous cancer cell populations