Baron Chloé S, Alemany Anna
Stem Cell Program and Division of Hematology/Oncology, Boston Children's Hospital and Dana-Farber Cancer Institute, Boston, MA, USA.
Stem Cell and Regenerative Biology Department, Harvard University, Cambridge, MA, USA.
Methods Mol Biol. 2025;2886:221-241. doi: 10.1007/978-1-0716-4310-5_11.
ScarTrace is a CRISPR/Cas9-based genetic lineage tracing method that allows for uniquely barcoding the DNA of single cells at a target GFP sequence during developing zebrafish embryos. Single cells from barcoded adult zebrafish can be isolated from various tissues (e.g., marrow, brain, eyes, fins), and their transcriptome and barcode sequences are captured by single-cell cDNA amplification and genomic DNA nested PCR, respectively. Computationally, cell type and barcode identification permit clone tracing and lineage tree reconstruction of tissues to unravel fate decisions during embryogenesis.
ScarTrace是一种基于CRISPR/Cas9的遗传谱系追踪方法,可在斑马鱼胚胎发育过程中对目标GFP序列处的单细胞DNA进行独特标记。可从条形码成年斑马鱼的各种组织(如骨髓、大脑、眼睛、鳍)中分离出单细胞,分别通过单细胞cDNA扩增和基因组DNA巢式PCR捕获它们的转录组和条形码序列。在计算上,细胞类型和条形码识别允许对组织进行克隆追踪和谱系树重建,以揭示胚胎发生过程中的命运决定。