Ying Yue, Yang Xingyu, Zhao Kai, Mao Jifang, Kuang Ying, Wang Zhugang, Sun Ruilin, Fei Jian
School of Life Sciences and Technology, Tongji University, Shanghai 200072, China.
Shanghai Research Center For Model Organisms, Shanghai 201210, China.
Nucleic Acids Res. 2015 Feb 18;43(3):1549-61. doi: 10.1093/nar/gkv016. Epub 2015 Jan 21.
The Krüppel-associated box (KRAB) domain is a transcription repression module from the largest family of transcriptional regulators encoded by higher vertebrates. We developed a drug-controllable regulation system based on an artificial KRAB-containing repressor (tTS) that targets the endogenous Hprt gene to explore the regulatory mechanism and molecular basis of KRAB-containing regulators within the context of an endogenous gene in vivo. We show that KRAB can mediate irreversible and reversible regulation of endogenous genes in mouse that is dependent on embryonic developmental stage. KRAB-induced stable DNA methylation within the KRAB binding region during the early embryonic stage, resulting in irreversible gene repression. In later stages, KRAB mainly induced de-acetylation and methylation of histone, resulting in reversible gene repression. Thus, we have characterized the KRAB-mediated regulation system within the context of an endogenous gene and multiple spatiotemporal ranges, thereby providing a basis for identifying the function of KRAB-containing regulators and aiding development of novel KRAB-based gene regulation tools in vivo.
克勒ppel相关框（KRAB）结构域是高等脊椎动物编码的最大转录调节因子家族中的一种转录抑制模块。我们基于一种靶向内源性Hprt基因的含人工KRAB的阻遏物（tTS）开发了一种药物可控调节系统，以在体内内源性基因的背景下探索含KRAB调节因子的调节机制和分子基础。我们表明，KRAB可介导小鼠内源性基因的不可逆和可逆调节，这取决于胚胎发育阶段。在胚胎早期阶段，KRAB诱导KRAB结合区域内稳定的DNA甲基化，导致不可逆的基因抑制。在后期阶段，KRAB主要诱导组蛋白的去乙酰化和甲基化，导致可逆的基因抑制。因此，我们在多个时空范围内的内源性基因背景下对KRAB介导的调节系统进行了表征，从而为鉴定含KRAB调节因子的功能以及协助开发基于KRAB的新型体内基因调节工具提供了基础。
