基因组区域的和编辑通过多重同质学导向的修复
Gregory M Findlay1, Evan A Boyle1, Ronald J Hause2
11] Department of Genome Sciences, University of Washington, Seattle, Washington 98195, USA [2].
Nature
|August 21, 2014
概括
和基因组编辑精确地映射了原生基因组内的DNA序列功能. 这种方法克服了以前技术的局限性,使得基因和调控元素的高分辨率功能分析成为可能.
科学领域:
- 遗传学 是一个遗传学.
- 分子生物学分子生物学
- 基因组学就是基因组学.
背景情况:
- 和突变是理解DNA序列的关键,但往往缺乏本地基因组背景.
- 目前使用情节或随机整合的方法限制了突变影响的解释性.
- 固有位点的背景对于准确评估序列功能至关重要.
研究的目的:
- 开发和展示一种直接在内源性基因组区域内进行和编辑的方法.
- 在特定的基因组位置精确测量所有可能的序列变异的功能影响.
- 提高基因变异的解释和基因组元素的功能剖析.
主要方法:
- 结合CRISPR/Cas9RNA引导的裂变与多重同质学导向的修复.
- 使用复杂的捐赠者模板库来对基因组区域进行和编辑.
- 在BRCA1外基因18和必需基因DBR1.1中进行和基因编辑.
主要成果:
- 在BRCA1.1中证明了用所有可能的六合体取代六基对区域.
- 在BRCA1.1中生成了所有可能的单核酸变异 (SNVs) 对于一个完整的外型.
- 由于无意中介衰变和拼接,观察到对BRCA1中转录丰度的显著影响.
- 在DBR1中测量了与功能影响相关的相对生长效应.
结论:
- 和基因组编辑提供了一种高分辨率的方法,用于在它们的原生环境中对DNA序列的功能剖析.
- 这种方法有助于研究 cis 调节元素,跨作用因子和临床变异解释.
- 能够精确测量多种突变的功能后果,改善基因组学理解.
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