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优化用于马原生质中的基因组编辑的细胞质基因编辑器

Ida Westberg¹, Frida Meijer Carlsen¹, Ida Elisabeth Johansen¹

¹Department of Plant and Environmental Sciences, Faculty of Science, The University of Copenhagen, Frederiksberg, Denmark.

Frontiers in genome editing

|September 18, 2023

概括

研究人员为土豆 (Solanum tuberosum) 开发了三种基编辑器 (CBEs),达到高达43%的C-to-T转化. 人类APOBEC3A (A3A) 在颗粒结合粉合成酶1基因中表现出最高的编辑效率.

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
基因编辑基因编辑

背景情况:

通过CRISPR/Cas9技术,可以在植物中进行基因编辑.
使用土豆U6促进体 (StU6) 的优化导向RNA (gRNA) 表达提高了Solanum tuberosum中的编辑效率.
基编辑器 (CBE) 是精确的C-T基转换的强大工具.

研究的目的:

为了生成和比较三个新的基基编辑器 (CBEs),专门为土豆 (Solanum tuberosum) 量身定制.
评估这些CBE在土豆品种Desiree中的颗粒结合粉合成酶1 (GBSS1) 基因中的C-to-T基编辑效率.

主要方法:

通过工程技术将Streptococcus pyogenes Cas9 (SpCas9) 转化为一个尼克酶 (nCas9).
与三种细胞因子除氨酶之一融合的nCas9:人类APOBEC3A (A3A),老鼠rAPOBEC1 (rA1) 或海CDA1 (CDA1),结合了 uracil-DNA糖酶抑制剂.
使用了优化的结构与土豆StU6促进器用于gRNA表达.

主要成果:

所有生成的CBE都在目标基因中表现出C-to-T基基编辑活动.
基于A3A的CBE表现出最高的整体效率,在所有等位基因中实现高达43%的C-T转换.
CDA1和rA1也显示出显著的编辑活动,rA1在C12位置无效.

结论:

量身定制的CBE,特别是A3A变体,对于精确的C-to-T基础编辑在土豆中非常有效.
这项研究为加速Solanum tuberosum的遗传改进提供了有价值的工具.
优化促进剂使用和除氨酶选择对于最大限度地提高植物中的CBE效率至关重要.

关键词:

在 U6 促销器中.细胞因子基编辑器基因组编辑基因组编辑原生促进体原生促进体这里是土豆土豆.一个原生质原生质.

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