Mesenchymal stem cells (MSCs) are of interest in regenerative medicine owing to their multilineage differentiation and self-renewal properties. Understanding the in vivo differentiation process is necessary for clinical applications including cell therapy and transplantation. This remains challenging owing to the lack of induction methods that imitate the natural programming process. Endogenous gene regulation of tissue-specific transcription factors is therefore desirable. In the present study, we demonstrated endogenous activation of adipogenic genes through the dCas9-based transcription system and achieved efficient induction of different types of adipocyte-like cells from MSCs. Interestingly, the MSCs converted via single-gene activation exhibited morphological and molecular properties of white adipocytes, while beige adipocyte-like cells were induced via multiplex gene activation of three specific transcription factors. These results reveal that the fate of MSCs can be effectively manipulated by direct activation of specific endogenous gene expression using a dCas9-based activator with reduced exogenous additives.

中文翻译：

---

使用CRISPR-Cas9通过内源基因激活控制间充质干细胞的成脂分化

间充质干细胞（MSC）由于其多系分化和自我更新特性而在再生医学中受到关注。了解体内分化过程对于包括细胞治疗和移植在内的临床应用是必不可少的。由于缺乏模仿自然编程过程的归纳方法，这仍然具有挑战性。因此，需要组织特异性转录因子的内源基因调节。在本研究中，我们通过基于dCas9的转录系统证明了脂肪形成基因的内源性激活，并成功诱导了来自MSC的不同类型的脂肪细胞样细胞的诱导。有趣的是，通过单基因激活转化的MSC表现出白色脂肪细胞的形态和分子特性，而米色脂肪细胞样细胞则通过三种特异性转录因子的多重基因激活而被诱导。