In early T-cell development, single cells dynamically shift expression of multiple transcription factors (TFs) during transition from multipotentiality to T-lineage commitment, but the functional roles of many TFs have been obscure. Here, synchronized in vitro differentiation systems, scRNA-seq with batch indexing, and controlled gene-disruption strategies have unraveled single-cell impacts of perturbing individual TFs at two stages in early T-cell development. Single-cell CRISPR perturbation revealed that early-acting TFs Bcl11a, Erg, Spi1 (PU.1), Gata3, and Tcf7 (TCF1) each play individualized roles promoting or retarding T-lineage progression and suppressing alternative trajectories, collectively determining population dynamics and path topologies. Later, during T-lineage commitment, cells prevented from expressing TF Bcl11b ‘realized’ this abnormality not with a developmental block, but by shifting into a divergent path via bZIP and Sox TF activation as well as E protein antagonism, finally exiting the T-lineage trajectory. These TFs thus exert a network of impacts to control progression kinetics, trajectories, and differentiation outcomes of early pro-T cells.

中文翻译：

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单细胞扰动剖析转录因子在早期 T 细胞发育中对进展速度和轨迹选择的控制

在早期 T 细胞发育中，单细胞在从多能性到 T 谱系承诺的转变过程中动态改变多种转录因子 (TF) 的表达，但许多 TF 的功能作用一直不清楚。这里，体外同步分化系统、带有批量索引的 scRNA-seq 和受控基因破坏策略已经阐明了在早期 T 细胞发育的两个阶段扰乱单个 TF 对单细胞的影响。单细胞 CRISPR 扰动显示早期作用的 TFs Bcl11a、Erg、Spi1 (PU.1)、Gata3 和 Tcf7 (TCF1) 各自发挥促进或延缓 T 谱系进展和抑制替代轨迹的个体化作用，共同决定种群动态和路径拓扑。后来，在 T 谱系定型期间，阻止表达 TF Bcl11b 的细胞“意识到”这种异常不是因为发育障碍，而是通过 bZIP 和 Sox TF 激活以及 E 蛋白拮抗转移到不同的路径，最终退出 T-血统轨迹。因此，这些 TF 会施加影响网络来控制进展动力学，