在Yarrowia lipolytica中使用CRISPR-Cas12a系统进行基因组编辑,转录调节和转发基因查
Adithya Ramesh1, Sangcheon Lee1, Ian Wheeldon2
1Department of Chemical and Environmental Engineering, University of California, Riverside, CA, USA.
Methods in molecular biology (Clifton, N.J.)
|March 12, 2024
概括
研究人员在Yarrowia lipolytica开发了CRISPR-Cas12a工具,用于基因组工程. 该系统能够实现基因淘汰,转录调节和功能性基因组选,增强合成生物学应用.
科学领域:
- 合成生物学 合成生物学
- 分子生物学分子生物学
- 生物技术是生物技术.
背景情况:
- 在基因组编辑方面,CRISPR-Cas12a系统比Cas9具有优势,包括自导RNA成熟和更短的表达盒.
- 雅罗维亚脂质菌 (Yarrowia lipolytica) 是一种具有重要生物技术潜力的工业相关的油性酵母菌.
- 目前对Y. lipolytica的基因组工程工具有限,这阻碍了其更广泛的应用.
研究的目的:
- 为Yarrowia lipolytica开发和验证基于CRISPR-Cas12a的合成生物学工具.
- 为了使多重基因淘汰,转录抑制和Y. lipolytica.中的激活.
- 为Y. lipolytica建立全基因组的CRISPR屏幕,以描述指导活动并预测高效的gRNAs.
主要方法:
- 为Y. lipolytica.开发一个CRISPR-Cas12a载体系统.
- 构建一个全基因组指南库,对Y. lipolytica.进行八倍覆盖.
- 执行聚合的CRISPR屏幕来评估指导活动.
- 机器学习的应用,以预测高效的Cas12a引导RNA.
主要成果:
- 成功地实现了CRISPR-Cas12a的基因淘汰,通过体形成.
- 证明了CRISPR-Cas12a与转录调节器融合,用于基因抑制和激活.
- 创建和验证一个全面的全基因组指南库,用于Y. lipolytica.
- 开发一种机器学习模型,用于预测高效的Cas12a gRNAs.
结论:
- 开发的CRISPR-Cas12a系统显著扩大了对Yarrowia lipolytica的合成生物学工具箱.
- 这些工具促进了简单的基因组工程,包括多重基因编辑和转录控制.
- 全基因组查方法和预测算法提高了Y. lipolytica.功能基因组研究的效率和范围.
关键词:
干扰和激活CRISPR的干扰和激活这就是CRISPR-Cas12a.基因组编辑 基因组编辑汇集的CRISPR屏幕是一个池.合成生物学 合成生物学转录法规 转录法规雅罗维亚 (Yarrowia lipolytica) 是一个高脂的植物.更多相关视频
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