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.

绘制体内细胞发育历史的经典方法依赖于需要复杂干预的技术，并且通常仅捕获单个轨迹或时间点。我们之前描述了一种发育条形码系统来解决这些问题，使用合成诱导的突变来记录有关每个细胞基因组中谱系的信息。该系统使用 MARC1 小鼠品系，该品系具有多个归巢引导 RNA，每个产生数百个突变等位基因，并结合产生指数多样性的条形码。在这里，我们详细介绍了公共存储库中提供的两条 MARC1 行。我们描述了使用 MARC1 小鼠的策略和实验设计注意事项。我们提供条形码检索和测序以及测序数据分析的协议。该协议根据小鼠中合成诱导的突变生成条形码，以实现谱系分析。