通过CRISPR引导的DNA聚合酶可以在可调节的窗口中实现所有核酸的多样化
Shakked O Halperin1,2,3, Connor J Tou1, Eric B Wong1
1Department of Bioengineering, University of California, Berkeley, Berkeley, CA, USA.
Nature
|August 3, 2018
概括
研究人员开发了EvolvR, 这种由CRISPR指导的DNA聚合酶工具可以快速编辑基因组,
科学领域:
- 分子生物学
- 遗传学
- 生物技术
背景情况:
- 遗传密码的多样化对于理解和设计生物系统至关重要.
- 现有的基因组多样化方法在某些生物系统和应用上存在局限性.
- 需要有效的向突变发生,加速进化和细胞系追踪方法.
研究的目的:
- 介绍EvolvR,一个用于用户定义基因组区域的持续多样化的新系统.
- 为了实现先进的遗传方法和期望的表型的快速进化.
- 通过条形码突变生成来促进细胞系的追踪.
主要方法:
- 开发了EvolvR,一个利用CRISPR引导的尼克酶来准工程DNA聚合酶的系统.
- 工程DNA聚合酶直接在用户定义的位置在可调节的窗口长度内产生突变.
- 确定了特定的尼克酶和聚合酶变体,以达到显著的突变率.
主要成果:
- 进化细胞的突变率高达野生细胞的77万倍.
- 该系统允许编辑多达350个核酸的窗口.
- 通过使用EvolvR成功鉴定出新型核糖体突变,使其具有抗谱素的特性.
结论:
- 通过CRISPR引导的DNA聚合酶使基因组位点的多重化和连续多样化成为可能.
- 它为基础研究和生物技术应用提供了强大的工具.
- 该系统对基因工程,进化研究和合成生物学具有广泛的用途.
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