從山中四因子OSKM中排除 Oct4 釋放 iPSC 的發(fā)展?jié)摿?br> Excluding Oct4 from Yamanaka Cocktail Unleashes the Developmental Potential of iPSCs - PubMed (nih.gov)
摘要
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.
Oct4 被廣泛認(rèn)為是四個(gè) Yamanaka 重編程因子中最重要的一個(gè)。在這里，我們展示了 Sox2、Klf4 和 cMyc (SKM) 的組合足以將小鼠體細(xì)胞重編程為誘導(dǎo)多能干細(xì)胞 (iPSC)。在成纖維細(xì)胞中同時(shí)誘導(dǎo) Sox2 和 cMyc 會(huì)立即觸發(fā)逆轉(zhuǎn)錄病毒沉默，這解釋了與先前研究的差異，這些研究試圖但未能使用逆轉(zhuǎn)錄病毒載體在沒有 Oct4 的情況下生成 iPSC。 SKM 誘導(dǎo)可以部分激活多能網(wǎng)絡(luò)，即使在 Oct4 敲除的成纖維細(xì)胞中也是如此。重要的是，在沒有外源 Oct4 的情況下重新編程會(huì)大大提高 iPSC 的發(fā)育潛力，這取決于它們?cè)谒谋扼w互補(bǔ)試驗(yàn)中產(chǎn)生全 iPSC 小鼠的能力。我們的數(shù)據(jù)表明，重編程過程中 Oct4 的過表達(dá)導(dǎo)致重編程過程中的脫靶基因激活和所得 iPSC 的表觀遺傳畸變，從而對(duì) iPSC 技術(shù)的進(jìn)一步開發(fā)和應(yīng)用具有重大意義。
