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Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing.
Nature Plants ( IF 15.8 ) Pub Date : 2019-07-08 , DOI: 10.1038/s41477-019-0459-z Tomohiko Kazama 1 , Miki Okuno 2 , Yuta Watari 3 , Shungo Yanase 3 , Chie Koizuka 4 , Yu Tsuruta 3 , Hajime Sugaya 3 , Atsushi Toyoda 5 , Takehiko Itoh 2 , Nobuhiro Tsutsumi 3 , Kinya Toriyama 1 , Nobuya Koizuka 4 , Shin-Ichi Arimura 3, 6
Nature Plants ( IF 15.8 ) Pub Date : 2019-07-08 , DOI: 10.1038/s41477-019-0459-z Tomohiko Kazama 1 , Miki Okuno 2 , Yuta Watari 3 , Shungo Yanase 3 , Chie Koizuka 4 , Yu Tsuruta 3 , Hajime Sugaya 3 , Atsushi Toyoda 5 , Takehiko Itoh 2 , Nobuhiro Tsutsumi 3 , Kinya Toriyama 1 , Nobuya Koizuka 4 , Shin-Ichi Arimura 3, 6
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Sequence-specific nucleases are commonly used to modify the nuclear genome of plants. However, targeted modification of the mitochondrial genome of land plants has not yet been achieved. In plants, a type of male sterility called cytoplasmic male sterility (CMS) has been attributed to certain mitochondrial genes, but none of these genes has been validated by direct mitochondrial gene-targeted modification. Here, we knocked out CMS-associated genes (orf79 and orf125) of CMS varieties of rice and rapeseed, respectively, using transcription activator-like effector nucleases (TALENs) with mitochondria localization signals (mitoTALENs). We demonstrate that knocking out these genes cures male sterility, strongly suggesting that these genes are causes of CMS. Sequencing revealed that double-strand breaks induced by mitoTALENs were repaired by homologous recombination, and that during this process, the target genes and surrounding sequences were deleted. Our results show that mitoTALENs can be used to stably and heritably modify the mitochondrial genome in plants.
中文翻译：
通过TALEN介导的线粒体基因组编辑来治疗细胞质雄性不育。
序列特异性核酸酶通常用于修饰植物的核基因组。但是，尚未实现对陆地植物线粒体基因组的靶向修饰。在植物中，一种称为细胞质雄性不育（CMS）的雄性不育已归因于某些线粒体基因，但这些基因均未通过直接线粒体基因靶向修饰得到验证。在这里，我们使用具有线粒体定位信号（mitoTALENs）的转录激活因子样效应核酸酶（TALENs）分别敲除了水稻和油菜籽CMS品种的CMS相关基因（orf79和orf125）。我们证明敲除这些基因可治愈男性不育症，强烈暗示这些基因是CMS的病因。测序表明，由mitoTALENs诱导的双链断裂可通过同源重组修复，并且在此过程中，目标基因和周围序列被删除。我们的结果表明，线粒体可用于稳定和遗传修饰植物中的线粒体基因组。
更新日期：2019-07-09
中文翻译：
通过TALEN介导的线粒体基因组编辑来治疗细胞质雄性不育。
序列特异性核酸酶通常用于修饰植物的核基因组。但是，尚未实现对陆地植物线粒体基因组的靶向修饰。在植物中，一种称为细胞质雄性不育（CMS）的雄性不育已归因于某些线粒体基因，但这些基因均未通过直接线粒体基因靶向修饰得到验证。在这里，我们使用具有线粒体定位信号（mitoTALENs）的转录激活因子样效应核酸酶（TALENs）分别敲除了水稻和油菜籽CMS品种的CMS相关基因（orf79和orf125）。我们证明敲除这些基因可治愈男性不育症，强烈暗示这些基因是CMS的病因。测序表明，由mitoTALENs诱导的双链断裂可通过同源重组修复，并且在此过程中，目标基因和周围序列被删除。我们的结果表明，线粒体可用于稳定和遗传修饰植物中的线粒体基因组。
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