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一个截断的内源U6促进器使在亚麻 (Linum usitatissimum L.) 中实现高效的CRISPR编辑

Feifei Li¹, Min Xue¹, Dongliang Guo¹

¹Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China.

Plants (Basel, Switzerland)

|April 12, 2025

概括

研究人员确定了用于CRISPR基因编辑的油麻中的功能性U6促进体. 亚麻U6-4促进剂表现出卓越的活性,提高了作物分子育种中的基因编辑效率.

科学领域:

植物分子生物学植物分子生物学
生物技术是生物技术.
遗传学遗传学是一个

背景情况:

在作物中的CRISPR基因编辑中,U6促进体至关重要.
识别内源性U6促进体可以提高基因编辑效率和分子育种中的精度.
油麻 (Linum usitatissimum L.) 是一种重要的石油生产作物.

研究的目的:

为了识别和表征在油亚麻中的功能性U6促进剂.
在使用内源性促进剂的油中建立高效的CRISPR/Cas9基因编辑系统.
为了比较亚麻U6促进者的编辑效率与阿拉比多普西斯U6促进者的编辑效率.

主要方法:

在油麻中识别和克隆四个U6 snRNA.
构建双露西法酶记者载体,用于促进体活性测试.
暂时转化为亚麻和尼科蒂亚纳 (Nicotiana benthamiana).
克里斯普尔/Cas9载体的构造和亚麻皮细胞的由Agrobacterium介导的转化.
对目标基因编辑频率的测序分析.

主要成果:

在油麻中发现了四种U6 snRNA与Arabidopsis thalianaU6 snRNA促进体具有高度同源性.
亚麻U6-4促进体表现出最高的转录活性,其最佳片段长度为342 bp.
由亚麻Lu14U6-4-5P促销器驱动的CRISPR/Cas9载体在LusPDS基因中显示出高于AtU6-P促销器的编辑频率.
转基因白色亚麻芽成功生成,表明基因编辑成功.

结论:

已确定的亚麻U6-4促进体是油亚麻中CRISPR/Cas9基因编辑的高度活跃和最佳促进体.
亚麻U6促进体驱动的CRISPR/Cas9系统在油亚麻中提供了增强的基因编辑效率,与使用Arabidopsis U6促进体的系统相比.
这项研究为推进分子育种和对油麻的基因改进提供了有价值的工具.

关键词:

这就是CRISPR/Cas9的作用.在U6的推广者.活动活动活动活动活动活动.亚麻 (Linum usitatissimum L.) 的使用情况基因记者基因记者截断截断是指切断.

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