Germ cell regeneration-mediated, enhanced mutagenesis in the ascidian Ciona intestinalis reveals flexible germ cell formation from different somatic cells.
Germ cell regeneration-mediated, enhanced mutagenesis in the ascidian Ciona intestinalis reveals flexible germ cell formation from different somatic cells.
复制标题
DOI：
10.1016/j.ydbio.2017.01.022
复制
发表时间：
2017-03
影响因子：
2.7
通讯作者：
Keita Yoshida;Akiko Hozumi;Nicholas Treen;Tetsushi Sakuma;Takashi Yamamoto;Maki Shirae‐Kurabayashi
中科院分区：
文献类型：
--
作者：
Keita Yoshida;Akiko Hozumi;Nicholas Treen;Tetsushi Sakuma;Takashi Yamamoto;Maki Shirae‐Kurabayashi
海鞘具有很高的再生能力，能够从体细胞中再生人工去除的原始生殖细胞（PGCs）。我们利用PGC再生建立了转录激活因子样效应核酸酶（TALEN）的生殖系诱变的有效方法。当从表达TALEN对的动物中人工去除PGC时，在靶基因中携带突变的体细胞转化为生殖细胞，该生殖细胞群体表现出比未去除PGC的动物更高的突变率。PGC再生使我们能够使用特定体细胞组织的TALEN表达载体进行生殖细胞诱变。出乎意料的是，表皮、神经组织和肌肉的顺式元件可用于生殖细胞诱变，表明再生的PGCs有多种来源，表明分化的Cionasomatic细胞恢复全能性的灵活性。单个雌雄同体的PGC再生动物的精子和卵子通常具有不同的突变，表明它们来自不同的细胞。PGC可以从体细胞中产生，即使母体PGC没有被去除，这表明PGC再生不仅仅是一个人工事件，而是可能具有内源性功能。这项研究提供了基因组编辑方法的技术创新，包括容易建立突变株系。此外，这项研究表明细胞机制和潜在的进化意义的PGC再生在玻璃海鞘。
The ascidianCiona intestinalishas a high regeneration capacity that enables the regeneration of artificially removed primordial germ cells (PGCs) from somatic cells. We utilized PGC regeneration to establish efficient methods of germ line mutagenesis with transcription activator-like effector nucleases (TALENs). When PGCs were artificially removed from animals in which a TALEN pair was expressed, somatic cells harboring mutations in the target gene were converted into germ cells, this germ cell population exhibited higher mutation rates than animals not subjected to PGC removal. PGC regeneration enables us to use TALEN expression vectors of specific somatic tissues for germ cell mutagenesis. Unexpectedly,ciselements for epidermis, neural tissue and muscle could be used for germ cell mutagenesis, indicating there are multiple sources of regenerated PGCs, suggesting a flexibility of differentiatedCionasomatic cells to regain totipotency. Sperm and eggs of a single hermaphroditic, PGC regenerated animal typically have different mutations, suggesting they arise from different cells. PGCs can be generated from somatic cells even though the maternal PGCs are not removed, suggesting that the PGC regeneration is not solely an artificial event but could have an endogenous function inCiona. This study provides a technical innovation in the genome-editing methods, including easy establishment of mutant lines. Moreover, this study suggests cellular mechanisms and the potential evolutionary significance of PGC regeneration inCiona.