TALEN-mediated shift of mitochondrial DNA heteroplasmy in MELAS-iPSCs with m.13513G>A mutation.
TALEN-mediated shift of mitochondrial DNA heteroplasmy in MELAS-iPSCs with m.13513G>A mutation.
复制标题
DOI：
10.1038/s41598-017-15871-y
复制
发表时间：
2017-11-14
影响因子：
4.6
通讯作者：
Hata R
中科院分区：
文献类型：
--
作者：
Yahata N;Matsumoto Y;Omi M;Yamamoto N;Hata R
诱导多能干细胞 (iPSC) 适用于研究由线粒体 DNA (mtDNA) 突变引起的线粒体疾病。在这里，我们从一名患有线粒体肌病、脑病、乳酸性酸中毒和中风样发作 (MELAS) 且具有 m.13513G>A 突变的患者中生成了 iPSC。对患者的真皮成纤维细胞进行了重新编程，我们建立了两个带有和不带有突变 mtDNA 的 iPSC 克隆。此外，我们尝试使用转录激活因子样效应核酸酶 (TALEN) 降低 iPSC 中突变 mtDNA 的水平。我们最初设计了铂TALEN，将其转运到线粒体中，识别包括m.13513位置的mtDNA序列，并优先切割G13513A突变体mtDNA（G13513A-mpTALEN）。通过转导 G13513A-mpTALEN，MELAS-iPSC 中的 m.13513G>A 异质性水平在短期内降低。我们的数据表明，这种靶向 mtDNA 的核酸酶将成为改变异质 iPSC 异质性水平的有力工具，这可能有助于阐明 mtDNA 突变引起的线粒体疾病的病理机制。
Induced pluripotent stem cells (iPSCs) are suitable for studying mitochondrial diseases caused by mitochondrial DNA (mtDNA) mutations. Here, we generated iPSCs from a patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) with the m.13513G>A mutation. The patient’s dermal fibroblasts were reprogrammed, and we established two iPSC clones with and without mutant mtDNA. Furthermore, we tried to decrease mutant mtDNA level in iPSCs using transcription activator-like effector nucleases (TALENs). We originally engineered platinum TALENs, which were transported into mitochondria, recognized the mtDNA sequence including the m.13513 position, and preferentially cleaved G13513A mutant mtDNA (G13513A-mpTALEN). The m.13513G>A heteroplasmy level in MELAS-iPSCs was decreased in the short term by transduction of G13513A-mpTALEN. Our data demonstrate that this mtDNA-targeted nuclease would be a powerful tool for changing the heteroplasmy level in heteroplasmic iPSCs, which could contribute to elucidation of the pathological mechanisms of mitochondrial diseases caused by mtDNA mutations.
登录
查看更多内容
影响因子：
3.5
作者：
Hance, N;Ekstrand, MI;Trifunovic, A
通讯作者：
Trifunovic, A
影响因子：
4.6
作者：
Sakuma, Tetsushi;Ochiai, Hiroshi;Kaneko, Takehito;Mashimo, Tomoji;Tokumasu, Daisuke;Sakane, Yuto;Suzuki, Ken-ichi;Miyamoto, Tatsuo;Sakamoto, Naoaki;Matsuura, Shinya;Yamamoto, Takashi
通讯作者：
Yamamoto, Takashi
影响因子：
12.4
作者：
Hashimoto, Masami;Bacman, Sandra R.;Moraes, Carlos T.
通讯作者：
Moraes, Carlos T.
影响因子：
--
作者：
Shanske, Sara;Coku, Jorida;DiMauro, Salvatore
通讯作者：
DiMauro, Salvatore
影响因子：
2.6
作者：
Kodaira, Masaki;Hatakeyama, Hideyuki;Yuasa, Shinsuke;Seki, Tomohisa;Egashira, Toru;Tohyama, Shugo;Kuroda, Yusuke;Tanaka, Atsushi;Okata, Shinichiro;Hashimoto, Hisayuki;Kusumoto, Dai;Kunitomi, Akira;Takei, Makoto;Kashimura, Shin;Suzuki, Tomoyuki;Yozu, Gakuto;Shimojima, Masaya;Motoda, Chikaaki;Hayashiji, Nozomi;Saito, Yuki;Goto, Yu-ichi;Fukuda, Keiichi
通讯作者：
Fukuda, Keiichi