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主编辑使用对联的pegRNAs准米中的NG-或NGG-PAM

Ayako Nishizawa-Yokoi¹, Keiko Iida^1,2, Akiko Mori¹

¹Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO), Tsukuba, Japan.

Frontiers in genome editing

|September 15, 2025

概括

主编辑 (PE) 提供精确的基因组编辑. 这项研究比较了野生类型的PE (PE-wt) 和使用工程基RNA的变体 (PE-NG),发现PE-wt即使在远端PAM中也有效,而PE-NG在大米中NGC-PAM位点的效率较低.

科学领域:

分子生物学分子生物学
基因组工程是基因组工程.
生物技术是生物技术.

背景情况:

主编辑 (PE) 促进了精确的基因组修改,包括基因替代,插入和删除.
带有RNA伪结序列的工程基RNA (epegRNAs) 增强了PE系统的稳定性.
现有的PE系统往往需要特定的protospacer相邻图案 (PAM) 配置,以限制目标站点的可访问性.

研究的目的:

为了比较基于野生型SpCas9的PE系统 (PE-wt) 和基于nSpCas9-NG变异的PE系统 (PE-NG) 使用配对的epegRNA的效率.
为了评估不同PAM配置 (NGG,NG) 和目标站点距离的PE效率.
评估PE-NG在大米中的适用性,特别是在NGC-PAM地点.

主要方法:

使用PE-wt与配对的epegRNAs准远端NGG-PAM.
使用PE-NG与配对的epegRNA准相邻的NG-PAM.
在使用PE系统的目标基因中引入了替代和删除突变.
在各种PAM类型 (NGG,CCN,NGA,NGT,NGC) 中,比较PE-wt和PE-NG之间的编辑频率.

主要成果:

在远端NGG-PAM位点,PE-wt证明了高效的基因组编辑.
在针对NGA和NGT-PAM站点时,PE-NG的效率与PE-wt相当.
与PE-wt相比,PE-NG在米NGC-PAM站点上的编辑效率明显较低.

结论:

对于基因组编辑来说,PE-wt是有效的,即使它针对的是远端PAM位点.
PE-NG扩大了PE的适用性,但在米中的特定NGC-PAM地点显示效率降低.
选择PE系统和PAM兼容性对于优化植物基因组编辑策略至关重要.

关键词:

这就是PAMAM.这就是SpCas9-NG.配对的基RNAs.首席编辑主要编辑米米饭米饭米饭米饭.

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