Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental Potential of iPSCs
Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental Potential of iPSCs
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DOI：
10.1016/j.stem.2019.10.002
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发表时间：
2019-12-05
期刊：
影响因子：
23.9
通讯作者：
Schoeler, Hans R.
中科院分区：
文献类型：
--
作者：
Velychko, Sergiy;Adachi, Kenjiro;Schoeler, Hans R.
Oct4被广泛认为是四种山中重编程因子中最重要的。本研究表明，Sox2、Klf4和cMyc （SKM）的组合足以将小鼠体细胞重编程为诱导多能干细胞（iPSCs）。在成纤维细胞中同时诱导Sox2和cMyc会立即触发逆转录病毒沉默，这解释了与先前使用逆转录病毒载体尝试但未能产生不含Oct4的iPSCs的研究的差异。SKM诱导可以部分激活多能性网络，甚至在oct4敲除成纤维细胞中也是如此。重要的是，在缺乏外源Oct4的情况下，重编程导致ipsc的发育潜力大大提高，这是由它们在四倍体互补实验中产生全ipsc小鼠的能力决定的。我们的数据表明，在重编程过程中过表达Oct4会导致重编程过程中的脱靶基因激活和由此产生的iPSC的表观遗传畸变，从而对iPSC技术的进一步开发和应用具有重要意义。
Oct4 is widely considered the most important among the four Yamanaka reprogramming factors. Here, we show that the combination of Sox2, Klf4, and cMyc (SKM) suffices for reprogramming mouse somatic cells to induced pluripotent stem cells (iPSCs). Simultaneous induction of Sox2 and cMyc in fibroblasts triggers immediate retroviral silencing, which explains the discrepancy with previous studies that attempted but failed to generate iPSCs without Oct4 using retroviral vectors. SKM induction could partially activate the pluripotency network, even in Oct4-knockout fibroblasts. Importantly, reprogramming in the absence of exogenous Oct4 results in greatly improved developmental potential of iPSCs, determined by their ability to give rise to all-iPSC mice in the tetraploid complementation assay. Our data suggest that overexpression of Oct4 during reprogramming leads to off-target gene activation during reprogramming and epigenetic aberrations in resulting iPSCs and thereby bear major implications for further development and application of iPSC technology.