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在西瓜和西瓜中提高CRISPR/Cas9的基因组编辑效率

Zhuanrong Wang¹, Lili Wan¹, Jian Ren¹

¹Institute of Crop, Wuhan Academy of Agricultural Sciences, Wuhan 430065, China.

|November 8, 2024

概括

在瓜子和西瓜中进行CRISPR基因编辑显示了改进的突变效率. 一个Csy4间距系统与一个Pol II促销器显著增强编辑,使大删除的特征改进.

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
遗传学是一个遗传学.

背景情况:

克里斯普尔/Cas9技术为改善作物特征提供了潜在的潜力.
在西瓜和西瓜等作物中提高CRISPR/Cas9突变效率至关重要,但具有挑战性.

研究的目的:

优化CRISPR/Cas9系统,以提高瓜子和西瓜的突变效率.
评估不同的sgRNA表达盒和促进器,以进行增强的基因组编辑.

主要方法:

设计了四个CRISPR/Cas9系统,使用不同的sgRNA表达盒准植物脱酶 (PDS) 基因.
利用Agrobacterium介导的转化进行基因传递.
在编辑的西瓜和西瓜幼苗中分析了表型和基因型变化.

主要成果:

在Pol II促进剂下使用tRNA和Csy4间隔器的CRISPR系统显著提高了瓜子的突变效率 (高达42.82%).
在78.95%的编辑西瓜和71.43%的编辑西瓜植物中,Csy4系统诱导了大片段删除 (LD).
在西瓜中使用Csy4系统实现了41.48%的PDS编辑效率.
鉴定了各种突变类型,包括异合体,三等位基因,嵌合体和同合体突变.

结论:

带有Csy4间隔器的sgRNA表达录音带驱动的Pol II促进器代表了提高瓜子和西瓜的CRISPR/Cas9编辑效率的高度有效策略.
这种优化的系统可促进显著的基因修改,包括大量的删除,以改善作物.

关键词:

克里斯普尔是什么意思?克里斯普尔是什么意思?这种植物名为Citrullus lanatus.黄瓜果 L. Cucumis melo L. 黄瓜果是什么意思这是Pol II的推广者.基因编辑基因编辑

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