功能增强剂-促进剂连接与CRISPR干扰的系统映射
Charles P Fulco1,2, Mathias Munschauer1, Rockwell Anyoha1
1Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
概括
研究人员使用CRISPR干扰来发现基因调节元件及其标,确定了控制MYC和GATA1表达的9种增强剂. 这种方法有助于理解基因网络和疾病的非编码变异.
科学领域:
- 基因组学和分子生物学
- 表观遗传学和基因调控
背景情况:
- 哺乳动物的基因表达依赖于非编码元素,但它们的功能和向基因在很大程度上是未知的.
- 了解这些元素对于解读生理过程和疾病机制至关重要.
研究的目的:
- 开发和应用高通量CRISPR干扰 (CRISPRi) 方法,以发现基因调控元素并识别它们的基因.
- 调查MYC和GATA1基本转录因子附近的监管因素.
主要方法:
- 为了高通量查,使用了集群定期间隔的短时间palindromic重复 (CRISPR) 干扰 (CRISPRi).
- 在MYC和GATA1附近对1兆基序列进行评估.
- 分析了染色体状态和染色体构成数据.
主要成果:
- 确定了9种调节基因表达和细胞增殖的远端增强剂.
- 通过定量色素特征区分的七种增强剂被发现可以调节MYC.
- 基于染色体和构成数据,开发了一种预测增强剂-促进剂连接的策略.
结论:
- 基于CRISPRi的方法有效地发现了调节元件及其向基因.
- 这种方法可以剖析复杂的转录网络.
- 它提供了一种解释非编码基因变异在人类疾病中的作用的方法.
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