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在土豆中通过mitoTALEN和mitoTALECD编辑线粒体DNA:编辑事件的分子特征和稳定性

Alessandro Nicolia¹, Nunzia Scotti², Nunzio D'Agostino³

¹CREA, Research Centre for Vegetable and Ornamental Crops, via Cavalleggeri 25, 84098, Pontecagnano, SA, Italy.

|January 5, 2024

概括

使用两种基于TALE的方法,即mitoTALEN和mitoTALECD,实现了土豆线粒体基因组编辑. 这两种方法都成功地修改了线粒体DNA (mtDNA),使这一重要的作物能够进行精确的遗传改变.

科学领域:

植物分子生物学植物分子生物学
基因组编辑技术的技术.
线粒体遗传学线粒体遗传学

背景情况:

研究新的基因组编辑工具用于作物改进.
在像土豆这样的植物繁殖作物中进行线粒体基因组编辑是未被充分探索的.
基于TALE的核酶和脱氨酶提供了精确的DNA修饰能力.

研究的目的:

评估和描述土豆线粒体中由mitoTALEN和mitoTALECD诱导的突变.
为了比较两个不同的线粒体基因组编辑方法的效率,准确性和稳定性.
为了在土豆线粒体基因组中实现orf125序列的目标淘汰.

主要方法:

使用基于TALE的双链断裂 (DSB) 诱导通过mitoTALEN.
通过mitoTALECD使用基于TALE的有针对性的基准编辑.
针对土豆线粒体基因组中的orf125序列进行编辑.

主要成果:

通过DSB修复,MitoTALEN诱导了向性淋巴体删除 (236-1066 bp).
MitoTALECD诱导了基替代,导致了无意义和错误的突变.
诱导的突变是同质细胞或异质细胞,同质细胞突变显示稳定的遗传.

结论:

线粒体和线粒体DNA都能够在高频率下对土豆进行精确的线粒体基因组修饰.
该研究提供了两种不同的编辑策略在相同的植物基因型和mtDNA区域的比较分析.
成功生成了具有稳定遗传的,精确修改的线粒体基因组的土豆植物.

关键词:

编辑基础编辑冠状体组 (chondriome) 是一种类型.双链断裂是双链断裂的原因之一.基因组编辑基因组编辑同类的重组组合.在MitoTALECDC中使用.这就是MitoTALEN.线粒体DNA编辑这种植物是Solanum tuberosum.

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