Cell lineage reprogramming via transgene overexpression of key master regulatory transcription factors has been well documented. However, the poor efficiency and lack of fidelity of this approach is problematic. Synthetic transcription factors (sTFs)—built from the repurposed CRISPR/Cas9 system—can activate endogenous target genes to direct differentiation or trigger lineage reprogramming. Here we explored whether sTFs could be used to steer mouse neural stem cells and mouse embryonic fibroblasts toward the oligodendrocyte lineage. We developed a non-viral modular expression system to enable stable multiplex delivery of pools of sTFs capable of transcriptional activation of three key oligodendrocyte lineage master regulatory genes (Sox10, Olig2, and Nkx6-2). Delivery of these sTFs could enhance neural stem cell differentiation and initiated mouse embryonic fibroblast direct reprograming toward oligodendrocyte progenitor-like cells. Our findings demonstrate the value of sTFs as tools for activating endogenous genes and directing mammalian cell-type identity.

通过转基因过表达关键主调控转录因子进行的细胞谱系重编程已得到充分记录。然而，这种方法存在效率低且缺乏保真度的问题。由重新调整用途的 CRISPR/Cas9 系统构建的合成转录因子 (sTF) 可以激活内源靶基因以指导分化或触发谱系重编程。在这里，我们探讨了 sTF 是否可用于引导小鼠神经干细胞和小鼠胚胎成纤维细胞向少突胶质细胞谱系发展。我们开发了一种非病毒模块化表达系统，能够稳定地多重递送 sTF 池，这些 sTF 池能够转录激活三个关键的少突胶质细胞谱系主调控基因（ Sox10 、 Olig2和Nkx6-2） 。这些 sTF 的传递可以增强神经干细胞分化，并启动小鼠胚胎成纤维细胞直接重编程为少突胶质祖细胞样细胞。我们的研究结果证明了 sTF 作为激活内源基因和指导哺乳动物细胞类型识别工具的价值。