Wang Lei, Xu Wenlong, Zhang Shun, Gundberg Gregory C, Zheng Christine R, Wan Zhengpeng, Mustafina Kamila, Caliendo Fabio, Sandt Hayden, Kamm Roger, Weiss Ron
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, MA, USA.
Nat Biomed Eng. 2024 Dec;8(12):1730-1743. doi: 10.1038/s41551-024-01229-z. Epub 2024 Jul 9.
Precisely sensing and guiding cell-state transitions via the conditional genetic activation of appropriate differentiation factors is challenging. Here we show that desired cell-state transitions can be guided via genetically encoded sensors, whereby endogenous cell-state-specific miRNAs regulate the translation of a constitutively transcribed endoribonuclease, which, in turn, controls the translation of a gene of interest. We used this approach to monitor several cell-state transitions, to enrich specific cell types and to automatically guide the multistep differentiation of human induced pluripotent stem cells towards a haematopoietic lineage via endothelial cells as an intermediate state. Such conditional activation of gene expression is durable and resistant to epigenetic silencing and could facilitate the monitoring of cell-state transitions in physiological and pathological conditions and eventually the 'rewiring' of cell-state transitions for applications in organoid-based disease modelling, cellular therapies and regenerative medicine.
通过适当分化因子的条件性基因激活来精确感知和引导细胞状态转变具有挑战性。在此，我们表明可以通过基因编码传感器来引导所需的细胞状态转变，即内源性细胞状态特异性微小RNA（miRNA）调节组成型转录的核糖核酸酶的翻译，而该核糖核酸酶又控制感兴趣基因的翻译。我们使用这种方法监测了几种细胞状态转变，富集了特定细胞类型，并通过内皮细胞作为中间状态自动引导人类诱导多能干细胞向造血谱系的多步分化。这种基因表达的条件性激活是持久的，并且对表观遗传沉默具有抗性，有助于在生理和病理条件下监测细胞状态转变，并最终在基于类器官的疾病建模、细胞治疗和再生医学应用中实现细胞状态转变的“重新布线”。
