B1 SINE-binding ZFP266 impedes reprogramming through suppression of chromatin opening mediated by pioneering factors
B1 SINE-binding ZFP266 impedes reprogramming through suppression of chromatin opening mediated by pioneering factors
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DOI：
10.1101/2022.01.04.474927
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发表时间：
2022-01
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影响因子：
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通讯作者：
Daniel F. Kaemena;Masahito Yoshihara;James Ashmore;Meryam Beniazza;Suling Zhao;Mårten Bertenstam;V. Olariu;S. Katayama;K. Okita;S. Tomlinson;K. Yusa;K. Kaji
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文献类型：
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作者：
Daniel F. Kaemena;Masahito Yoshihara;James Ashmore;Meryam Beniazza;Suling Zhao;Mårten Bertenstam;V. Olariu;S. Katayama;K. Okita;S. Tomlinson;K. Yusa;K. Kaji
诱导多能干细胞重编程本身是低效的，理解这种低效的分子机制是成功控制细胞身份的关键。在这里，我们报告了通过CRISPR/ cas9介导的全基因组敲除（KO）筛选鉴定出的16个新的重编程障碍基因。其中，在几种iPSC重编程设置中，预测的KRAB锌指蛋白（KRAB- zfp） Zfp266的缺失强烈且持续地增强了iPSC的生成，成为最强大的障碍。进一步分析发现，ZFP266结合OCT4 （POU5F1）、SOX2和KLF4等先导因子结合位点附近的短穿插核元件（Short Interspersed Nuclear element, SINEs），阻碍染色质打开。用共激活域取代KRAB共抑制子将ZFP266从重编程抑制剂转变为有效的重编程促进者。这项工作提出了sin - krab - zfp相互作用是染色质可及性增强子的关键调节因子，可以有效地改变细胞身份，也可以作为进一步阐明阻碍重编程的分子机制的资源。
Induced pluripotent stem cell reprogramming is inherently inefficient and understanding the molecular mechanisms underlying this inefficiency holds the key to successfully control cellular identity. Here, we report 16 novel reprogramming roadblock genes identified by CRISPR/Cas9-mediated genome-wide knockout (KO) screening. Of these, depletion of the predicted KRAB zinc finger protein (KRAB-ZFP) Zfp266 strongly and consistently enhanced iPSC generation in several iPSC reprogramming settings, emerging as the most robust roadblock. Further analyses revealed that ZFP266 binds Short Interspersed Nuclear Elements (SINEs) adjacent to binding sites of pioneering factors, OCT4 (POU5F1), SOX2 and KLF4, and impedes chromatin opening. Replacing the KRAB co-suppressor with a co-activator domain converted ZFP266 from a reprogramming inhibitor to a potent reprogramming facilitator. This work proposes SINE-KRAB-ZFP interaction to be a critical regulator of chromatin accessibility at enhancers for efficient cellular identity changes and also serves as a resource to further illuminate molecular mechanisms hindering reprogramming.