在活细胞中,基因编码的光记者对活细胞中的基因素甲基化有影响
Chi-Wang Lin1, Cindy Y Jao, Alice Y Ting
1Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Journal of the American Chemical Society
|May 13, 2004
概括
研究人员开发了新的光记者来跟踪基因组甲基化,这是基因调节的关键过程. 这些工具可以精确监测活细胞中的基因沉默和X染色体失活.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
背景情况:
- 基因组甲基化是调节基因表达的关键表观遗传修饰.
- 现有的检测基因组甲基化的方法在活细胞中缺乏空间和时间分辨率.
研究的目的:
- 设计和表征新的基因编码光报道器用于基因素H3甲基化.
- 为了实时监测完整细胞中的表观遗传修饰.
主要方法:
- 构建四部分化学蛋白质的结构,这些蛋白质融合了组素H3,染色体 (HP1,Polycomb) 和FRET对 (CFP,YFP).
- 使用光共振能量转移 (FRET) 在体外和单个活细胞中对酶甲基化反应的记者反应的表征.
主要成果:
- 开发出响应于基因素H3氨酸9和氨酸27的甲基化反应的记者.
- 在甲基化后观察到YFP/CFP排放率的显著增加 (分别为60%和28%).
- 在体外试验和单个活细胞中都表现出报告器功能.
结论:
- 开发的光报道器为研究组织蛋白甲基化提供了强大的工具.
- 这些记者为基因沉默和X染色体失活的高分辨率调查提供便利.
- 该系统可能有助于发现基因素脱甲基酶活性.
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