Human pluripotent stem cells (hPSCs) offer an unprecedented opportunity to model diverse cell types and tissues. To enable systematic exploration of the programming landscape mediated by transcription factors (TFs), we present the Human TFome, a comprehensive library containing 1,564 TF genes and 1,732 TF splice isoforms. By screening the library in three hPSC lines, we discovered 290 TFs, including 241 that were previously unreported, that induce differentiation in 4 days without alteration of external soluble or biomechanical cues. We used four of the hits to program hPSCs into neurons, fibroblasts, oligodendrocytes and vascular endothelial-like cells that have molecular and functional similarity to primary cells. Our cell-autonomous approach enabled parallel programming of hPSCs into multiple cell types simultaneously. We also demonstrated orthogonal programming by including oligodendrocyte-inducible hPSCs with unmodified hPSCs to generate cerebral organoids, which expedited in situ myelination. Large-scale combinatorial screening of the Human TFome will complement other strategies for cell engineering based on developmental biology and computational systems biology.

人类多能干细胞 (hPSC) 为模拟不同的细胞类型和组织提供了前所未有的机会。为了能够系统地探索转录因子 (TF) 介导的编程景观，我们提出了 Human TFome，这是一个包含 1,564 个 TF 基因和 1,732 个 TF 剪接亚型的综合文库。通过筛选三个 hPSC 系中的文库，我们发现了 290 个 TF，其中包括 241 个之前未报道的 TF，它们在 4 天内诱导分化，且不改变外部可溶性或生物力学线索。我们使用其中的四个命中将 hPSC 编程为神经元、成纤维细胞、少突胶质细胞和血管内皮样细胞，这些细胞与原代细胞具有分子和功能相似性。我们的细胞自主方法能够同时将 hPSC 并行编程为多种细胞类型。我们还通过将少突胶质细胞诱导型 hPSC 与未修饰的 hPSC 纳入其中来生成大脑类器官，从而证明了正交编程，从而加速了原位髓鞘形成。人类 TFome 的大规模组合筛选将补充基于发育生物学和计算系统生物学的其他细胞工程策略。