Wang Liu, Zheng Aihua, Yi Ling, Xu Chongren, Ding Mingxiao, Deng Hongkui
Department of Cell Biology and Genetics, College of Life Science, Peking University, Beijing, PR China.
Biochem Biophys Res Commun. 2004 Dec 3;325(1):302-7. doi: 10.1016/j.bbrc.2004.10.028.
Nuclear reprogramming is critical for animal cloning and stem cell creation through nuclear transfer, which requires extensive remodeling of chromosomal architecture involving dramatic changes in chromatin-binding proteins. To understand the mechanism of nuclear reprogramming, it is critical to identify chromatin-binding factors specify the reprogramming process. In this report, we have developed a high-throughput selection method, based on T7 phage display and chromatin immunoprecipitation, to isolate chromatin-binding factors expressed in mouse embryonic stem cells using primary mouse embryonic fibroblast chromatin. Seven chromatin-binding proteins have been isolated by this method. We have also isolated several chromatin-binding proteins involved in hepatocyte differentiation. Our method provides a powerful tool to rapidly and selectively identify chromatin-binding proteins. The method can be used to study epigenetic modification of chromatin during nuclear reprogramming, cell differentiation, and transdifferentiation.
核重编程对于通过核移植进行动物克隆和干细胞创建至关重要，这需要对染色体结构进行广泛重塑，涉及染色质结合蛋白的显著变化。为了理解核重编程的机制，关键是要鉴定出指定重编程过程的染色质结合因子。在本报告中，我们基于T7噬菌体展示和染色质免疫沉淀开发了一种高通量筛选方法，以使用原代小鼠胚胎成纤维细胞染色质分离在小鼠胚胎干细胞中表达的染色质结合因子。通过这种方法已分离出七种染色质结合蛋白。我们还分离出了几种参与肝细胞分化的染色质结合蛋白。我们的方法提供了一种强大的工具，可快速、选择性地鉴定染色质结合蛋白。该方法可用于研究核重编程、细胞分化和转分化过程中染色质的表观遗传修饰。
