Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
Spatial Genomics, Inc., Pasadena, CA 91106, USA.
Science. 2021 Apr 9;372(6538). doi: 10.1126/science.abb3099.
During multicellular development, spatial position and lineage history play powerful roles in controlling cell fate decisions. Using a serine integrase-based recording system, we engineered cells to record lineage information in a format that can be read out in situ The system, termed integrase-editable memory by engineered mutagenesis with optical in situ readout (intMEMOIR), allowed in situ reconstruction of lineage relationships in cultured mouse cells and flies. intMEMOIR uses an array of independent three-state genetic memory elements that can recombine stochastically and irreversibly, allowing up to 59,049 distinct digital states. It reconstructed lineage trees in stem cells and enabled simultaneous analysis of single-cell clonal history, spatial position, and gene expression in brain sections. These results establish a foundation for microscopy-readable lineage recording and analysis in diverse systems.
在多细胞发育过程中，空间位置和谱系历史在控制细胞命运决策方面起着强大的作用。我们使用基于丝氨酸整合酶的记录系统，设计细胞以记录谱系信息的格式，可以在原位读出。该系统通过光原位读出的工程诱变被称为整合酶可编辑记忆（integrase-editable memory by engineered mutagenesis with optical in situ readout，intMEMOIR），允许在培养的小鼠细胞和果蝇中原位重建谱系关系。intMEMOIR 使用独立的三态遗传记忆元件阵列，可以随机和不可逆地重组，允许多达 59049 个不同的数字状态。它在干细胞中重建了谱系树，并能够同时分析脑切片中单细胞克隆历史、空间位置和基因表达。这些结果为在不同系统中进行显微镜可读的谱系记录和分析奠定了基础。
