Lineage Analysis at Single-Cell Resolution by Twin-Spot MARCM with Lineage-Restricted Drivers

Cell lineage analysis is primarily undertaken to understand cell fate specification and diversification along a cell lineage tree. Built with dual repressible markers, twin-spot mosaic analysis with repressible cell markers (MARCM) labels the two daughter cells made by a common precursor in distinct colors. The power of twin-spot MARCM to systematically subdivide complex lineages is exemplified in studies of Drosophila neural stem-cell lineages. To achieve targeted lineage analysis, lineage-restricted drivers can be created by restricting and immortalizing gene expression to a lineage of interest. Here, we describe detailed methods to perform lineage analysis in Drosophila neural stem-cell lineages utilizing lineage-specific drivers. Our innovative lineage tracing method can help resolve complex tissue development.