Lineage barcoding in mice with homing CRISPR.

Kathleen Leeper, Kian Kalhor, Andyna Vernet, Amanda Graveline, George M Church, Prashant Mali, Reza Kalhor

Nature protocols 2021 Apr

filter terms:

Classic approaches to mapping the developmental history of cells in vivo have relied on techniques that require complex interventions and often capture only a single trajectory or moment in time. We have previously described a developmental barcoding system to address these issues using synthetically induced mutations to record information about each cell's lineage in its genome. This system uses MARC1 mouse lines, which have multiple homing guide RNAs that each generate hundreds of mutant alleles and combine to produce an exponential diversity of barcodes. Here, we detail two MARC1 lines that are available from a public repository. We describe strategies for using MARC1 mice and experimental design considerations. We provide a protocol for barcode retrieval and sequencing as well as the analysis of the sequencing data. This protocol generates barcodes based on synthetically induced mutations in mice to enable lineage analysis.

Citation

Kathleen Leeper, Kian Kalhor, Andyna Vernet, Amanda Graveline, George M Church, Prashant Mali, Reza Kalhor. Lineage barcoding in mice with homing CRISPR. Nature protocols. 2021 Apr;16(4):2088-2108