相关实验视频
Updated: Jan 19, 2026
09:20
CRISPR-Mediated Reorganization of Chromatin Loop Structure
Published on: September 14, 2018
13.1K
染色体循环挤出在生理V(D) J重组中的基本作用
Yu Zhang1,2,3,4,4, Xuefei Zhang1,2,3, Zhaoqing Ba1,2,3
1Program in Cellular and Molecular Medicine, Boston Children's Hospital, Boston, MA, USA.
Nature
|September 13, 2019
概括
在RAG内核酶
科学领域:
- 免疫学
- 分子生物学
- 遗传学
背景情况:
- 在B细胞原体中,RAG内核酶启动免疫球蛋白重链 (Igh) V(D) J组合.
- 通过CTCF结合元件 (CBEs) 固定在Igh的3'端的染色素环域在这个过程中发挥作用.
- 这些域内的重组中心 (RC) 包含关键元素,包括D和JH段以及增强剂.
研究的目的:
- 调查RAG内核酶在Igh V(D) J重组中的扫描作用.
- 确定染色体定向和染色质结构如何影响D和JH段的RAG介导连接.
- 阐明DQ52在重组中心的结合机制.
主要方法:
- 使用基于等离子体的测试来研究RAG内核酶活性.
- 使用核酶死Cas9阻碍RAG扫描.
- 在重组中心进行高分辨率的染色体相互作用映射.
主要成果:
- JH-23RSS的染色体定向指导RAG扫描上游染色体的D-12RSS,调解大多数D段的删除连接.
- DQ52段通过与JH段独立的扩散相关机制连接.
- 阻碍RAG扫描,如Cas9结合或转录,焦点上增加RAG活动,并与循环挤出中断有关.
结论:
- RAG内核酶扫描由染色体定向编程,并受染色体循环动态的影响.
- D段连接的机制有所不同,DQ52采用了不同的基于扩散的途径.
- 染色体结构和RAG扫描是密切相关的,扫描的障碍影响RAG准和V(D) J组合.
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