Single-cell RNAseq of Brachyury knockout chimeric mouse embryos at embryonic days 8.5 and 7.5 of mouse development.. Single-cell RNAseq of Brachyury knockout chimeric mouse embryos at embryonic days 8.5 and 7.5 of mouse development.

Chimeric embryos were generated to investigate the effect of T knockout in mouse embryos by single-cell RNA-sequencing. T is an essential transcription factor for axial embryonic patterning. Chimeric embryos contain tissue that is T+/+, which prevents global developmental failures. Embryos were generated by blastocyst injection of tdTomato-labelled, Tal1-/- mouse embryonic stem cells into wild type embryos. After blastocyst harvest, cells were flow-sorted before 10X Genomics library preparation and single-cell RNA-sequencing.

本研究构建嵌合胚胎，通过单细胞RNA测序（single-cell RNA-sequencing）探究T基因敲除（T knockout）对小鼠胚胎的影响。T基因是调控轴式胚胎模式形成的关键转录因子。此类嵌合胚胎携带T基因野生型（T+/+）的组织，可避免整体发育失败。实验通过将经tdTomato标记的Tal1基因敲除（Tal1-/-）小鼠胚胎干细胞注射至野生型小鼠囊胚中，构建目标胚胎。在收集囊胚后，研究人员先通过流式细胞分选分离目标细胞，随后开展10X Genomics文库制备及单细胞RNA测序工作。