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有效的CRISPR/Cas基因编辑在由棉花叶病毒诱导的棉花中

Jiajun Zhang¹, Peihong Dai², Zheng Weng¹

¹College of Agriculture, Xinjiang Agricultural University, 311 Nongda East Road, Urumqi 830052, PR China.

Journal of biotechnology

|October 11, 2025

概括

这项研究开发了一种高效的CRISPR/Cas基因编辑系统在棉花中使用棉叶纹病毒 (CLCrV) 作为传递载体,用于多重基因编辑和腺基编辑.

科学领域:

植物生物技术植物生物技术
分子生物学分子生物学
遗传学遗传学是一个

背景情况:

植物病毒载体提供自主复制和细胞间传播,非常适合提供CRISPR/Cas基因编辑工具.
克里斯普尔/卡斯系统对于精确的植物基因组修改具有强大作用.

研究的目的:

构建和评估一个使用棉叶结病毒 (CLCrV) 作为传递载体的棉花CRISPR/Cas系统.
评估CLCrV介导的基因淘汰和棉花中的腺基编辑的效率.
探索同时进行多目标基因编辑的能力.

主要方法:

在棉花中构建一个CLCrV介导的CRISPR/Cas系统.
用针对 GhAGL16, GhPDS 和 GhCLA1.1 的 sgRNAs 接种棉线 (Pro35s::Cas9 和 ProUbi::Cas9) 的疫苗.
探索聚合病毒注射用于同时多目标编辑.
使用过度表达nCas9-TadA7.10的棉线进行腺基编辑.
突变检测和深度测序用于效率和特异性分析.

主要成果:

在Pro35s::Cas9和ProUbi::Cas9棉线上,编辑效率与CLCrV-Cas9系统相比较.
通过聚合的CLCrV-sgRNA注射,同时进行多目标基因编辑是可行的,尽管效率略有降低.
通过CLCrV介导的腺基编辑,在棉花GhPEBP中实现了特定的A-to-G转换.

结论:

已经建立了一个高效的CLCrV介导的CRISPR系统用于棉花基因编辑.
这个系统支持多重基因淘汰和特定的腺基编辑.
开发的系统为棉花的先进基因操纵提供了一个多功能工具.

关键词:

编辑ABE基础编辑棉花棉花棉花棉花棉花棉花叶纹病毒病毒多重编辑多重编辑.病毒池是一个病毒池.

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