Bowling Sarah, Camargo Fernando D
Stem Cell Program, Boston Children's Hospital, Boston, MA, USA.
Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA.
Methods Mol Biol. 2025;2886:281-298. doi: 10.1007/978-1-0716-4310-5_14.
The CRISPR-activated repair lineage tracing (CARLIN) mouse line uses DNA barcoding to enable high-resolution tracing of cell lineages in vivo (Bowling et al, Cell 181, 1410-1422.e27, 2020). CARLIN mice contain expressed barcodes that allow simultaneous interrogation of lineage and gene expression information from single cells. Furthermore, barcode editing is fully inducible, resulting in cell lineage labeling that can be performed at any time point in development or adulthood. This chapter details the protocols followed for maintaining CARLIN mice, inducing barcoding, and amplifying the CARLIN barcode from DNA, RNA, and single-cell RNA-sequencing libraries for next-generation sequencing.
CRISPR激活修复谱系追踪（CARLIN）小鼠品系利用DNA条形码技术在体内实现对细胞谱系的高分辨率追踪（Bowling等人，《细胞》，第181卷，第1410 - 1422.e27页，2020年）。CARLIN小鼠包含已表达的条形码，可同时从单细胞中获取谱系和基因表达信息。此外，条形码编辑是完全可诱导的，从而能够在发育或成年期的任何时间点进行细胞谱系标记。本章详细介绍了饲养CARLIN小鼠、诱导条形码生成以及从DNA、RNA和单细胞RNA测序文库中扩增CARLIN条形码用于下一代测序所遵循的方案。
