Controlling gene activation by enhancers through a drug-inducible topological insulator.
Controlling gene activation by enhancers through a drug-inducible topological insulator.
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通过药物诱导的拓扑绝缘体通过增强子控制基因激活。
DOI：
10.1101/534073
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发表时间：
2019
期刊：
影响因子：
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通讯作者：
Hishikawa K
中科院分区：
文献类型：
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作者：
Tsujimura T;Takase O;Yoshikawa M;Sano E;Hayashi M;Hoshi K;Takato T;Toyoda A;Okano H;Hishikawa K
尽管人们对基因-增强子相互作用的调控进行了深入的研究，但其应用仍然有限。在这里，我们重组了CTCF结合位点的阵列，并设计了一种用于染色质工程(SITCH)的合成拓扑绝缘体。通过将SITCH与连接到KRAB结构域的TetR偶联来诱导异染色质并使绝缘失效，我们开发了一个药物诱导系统来控制增强子对基因的激活。在人类诱导的多能干细胞中，在MYC和增强子下调MYC之间插入SING。SINIT的进行性突变导致基因-增强子相互作用的优先升级，证实了SING强大的绝缘能力。斯蒂奇还改变了MYC周围的表观遗传状态。药物诱导的时程分析发现H3K27me3抑制标记的沉积和去除跟随并反映了表达变化，但不在表达变化之前和确定之前。最后，在NEUROG2附近缝合削弱了神经前体细胞分化过程中的基因激活。因此，Stich应该在功能遗传学研究中有广泛的用途。
While regulation of gene-enhancer interaction is intensively studied, its application remains limited. Here, we reconstituted arrays of CTCF-binding sites and devised asynthetictopologicalinsulator withtetO forchromatin-engineering (STITCH). By coupling STITCH with tetR linked to the KRAB domain to induce heterochromatin and disable the insulation, we developed a drug-inducible system to control gene activation by enhancers. In human induced pluripotent stem cells, STITCH inserted betweenMYCand the enhancer down-regulatedMYC.Progressive mutagenesis of STITCH led to a preferential escalation of the gene-enhancer interaction, corroborating the strong insulation ability of STITCH. STITCH also altered epigenetic states aroundMYC. Time-course analysis by drug induction uncovered deposition and removal of H3K27me3 repressive marks follows and reflects, but does not precede and determine, the expression change. Finally, STITCH inserted nearNEUROG2impaired the gene activation in differentiating neural progenitor cells. Thus, STITCH should be broadly useful for functional genetic studies.