A Method for Precisely Identifying Modifications to Plant Mitochondrial Genomes by mitoTALENs
A Method for Precisely Identifying Modifications to Plant Mitochondrial Genomes by mitoTALENs
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一种通过 mitoTALEN 精确识别植物线粒体基因组修饰的方法
DOI：
10.1007/978-1-0716-2922-2_25
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发表时间：
2023
影响因子：
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通讯作者：
Arimura Shin-ichi
中科院分区：
文献类型：
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作者：
Kazama Tomohiko;Arimura Shin-ichi
转化植物线粒体基因组的能力具有许多益处。尽管目前将外源DNA递送至线粒体非常困难，但现在可以使用靶向线粒体的转录激活因子样效应物核酸酶（mitoTALEN）敲除线粒体基因。这种敲除已经通过将mitoTALEN编码基因遗传转化到核基因组中来实现。先前的研究表明，由mitoTALEN诱导的双链断裂（DSB）通过异位同源重组修复。作为通过同源重组进行DNA修复的结果，含有mitoTALEN靶位点的基因组的一部分被缺失。缺失和修复过程导致线粒体基因组变得更加复杂。在这里，我们描述了一种方法，用于确定异位同源重组事件后发生的mitoTALENs诱导的双链断裂的修复。
The ability to transform plant mitochondrial genomes has many benefits. Although delivery of foreign DNA to mitochondria is presently very difficult, it is now possible to knock out mitochondrial genes using mitochondria-targeted transcription activator-like effector nucleases (mitoTALENs). Such knockouts have been achieved by a genetic transformation of mitoTALENs encoding genes into the nuclear genome. Previous studies have shown that double-strand breaks (DSBs) induced by mitoTALENs are repaired by ectopic homologous recombination. As a result of DNA repair by homologous recombination, a portion of the genome containing the mitoTALEN target site is deleted. The deletion and repair process cause the mitochondrial genome to become more complex. Here, we describe a method for identifying the ectopic homologous recombination events that occur following the repair of double-strand breaks induced by mitoTALENs.