Zic3 Enhances the Generation of Mouse Induced Pluripotent Stem Cells
Zic3 Enhances the Generation of Mouse Induced Pluripotent Stem Cells
复制标题
DOI：
10.1089/scd.2012.0651
复制
发表时间：
2013-07-01
影响因子：
4
通讯作者：
Kumar, Anujith
中科院分区：
文献类型：
--
作者：
Declercq, Jeroen;Sheshadri, Preethi;Kumar, Anujith
小脑锌指蛋白(Zic)3是Gli转录因子家族的成员，是维持胚胎干细胞(ESCs)多能性所必需的，已有报道以Oct4/Sox2不依赖的方式激活TF Nanog。此前，我们发现Zic3(Z)与Yamanka因子OCT4、SOX2和KLF4(OSK)相结合，可以从人成纤维细胞诱导出神经前体细胞。然而，与单独使用OSK相比，在小鼠胚胎成纤维细胞中转导的TFS(OSKZ)类似的组合可促进诱导多能干细胞(IPSCs)的形成，但不能促进神经前体细胞的形成。OSKZ来源的IPSCs在集落形态、碱性磷酸酶和多能性基因的表达、类胚体和畸胎瘤形成方面与mESCs没有区别。荧光素酶启动子实验证明，ZIC3激活了关键的多能性调节因子Nanog的转录。在IPSC衍生的过程中，观察到Zic3介导的Nanog和TBX3基因的表达增强，这是已知的促进iPSC衍生的基因。Zic3不仅提高了重编程效率，而且内源Zic3蛋白的重新激活对于IPSC的生成是必不可少的，因为在用OSKM生成iPSC的过程中敲除Zic3显著减少了克隆数量。综上所述，我们的结果揭示了Zic3在小鼠ipscs生成中的重要作用。
Zinc finger protein of the cerebellum (Zic)3, a member of Gli family of transcription factors (TFs), is essential for maintaining pluripotency of embryonic stem cells (ESCs) and has been reported to activate TF Nanog in an Oct4/Sox2-independent manner. Previously, we showed that Zic3 (Z), in combination with the Yamanka factors OCT4, SOX2, and KLF4 (OSK), induces neural progenitor-like cells from human fibroblasts. However, a similar combination of TFs (OSKZ) transduced in mouse embryonic fibroblasts resulted in enhanced induced pluripotent stem cells (iPSCs) formation compared with OSK alone, but not neuroprogenitors. OSKZ-derived iPSCs are indistinguishable from mESCs in colony morphology, expression of alkaline phosphatase and pluripotency genes, and embryoid body and teratoma formation. Zic3 activates the transcription of Nanog, a key pluripotency regulator, as evidenced by a luciferase promoter assay. During the course of iPSC derivation, Zic3-mediated enhanced expression of Nanog and Tbx3, gene known to enhance iPSCs derivation, is observed. Not only does Zic3 enhance the reprogramming efficiency, but also reactivation of the endogenous Zic3 protein is essential for the generation of iPSCs, as knockdown of Zic3 during the iPSC generation with OSKM significantly reduced the number of colonies. Together, our result uncovers an important role of Zic3 in generating mouse iPSCs.