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优化大白菜 (Brassica oleracea) 的种植方式 L.) 原生体转化用于使用CRISPR/Cas9进行基因组编辑

Ester Stajič¹, Urban Kunej¹

¹Department of Agronomy, Biotechnical Faculty, University of Ljubljana, Ljubljana, Slovenia.

Frontiers in plant science

|October 18, 2023

概括

优化CRISPR/Cas9传递到白菜原生体中可以提高基因组编辑效率. 关键因素,如PEG4000度和化时间,为成功的作物改良和基因功能研究进行了微调.

科学领域:

农业科学农业科学
分子生物学分子生物学
生物技术是生物技术.

背景情况:

使用集群定期间隔的短Palindromic重复/CRISPR相关系统 (CRISPR/Cas9) 的基因组编辑对于作物改进至关重要.
为了成功编辑基因组,有效地将CRISPR/Cas9输入植物细胞,特别是原生质细胞,至关重要.

研究的目的:

为了优化CRISPR/Cas9向量向白菜原塑体的输送.
确定影响转换效率和编辑结果的关键因素.
建立一个可靠的基因组编辑协议在brassicas.

主要方法:

评估聚乙烯糖4000 (PEG4000) 度,潜伏时间和CRISPR/Cas9输送的等离子体量.
使用安普利康序列测定来确认目标突变.
在转化后评估原生质的生存能力.

主要成果:

优化的转换协议实现了26.4%的编辑效率.
证实了PEG4000度和潜伏时间对突变率的显著影响.
尽管进行了优化参数,但原生质细胞的高活力仍然存在.

结论:

优化的协议使白菜原生体中的高效基因组编辑成为可能.
这些发现支持基因功能分析和brassicas中的亚细胞局部化研究.
这项研究为推进作物遗传改进提供了有价值的方法.

关键词:

B. oleracea 这种植物这就是CRISPR/Cas9的作用.通过PEG介导的转化过程.基因组编辑基因组编辑原始塑料是原始塑料中的一个.

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