Repeating pattern of non-RVD variations in DNA-binding modules enhances TALEN activity.
Repeating pattern of non-RVD variations in DNA-binding modules enhances TALEN activity.
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DOI：
10.1038/srep03379
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发表时间：
2013-11-29
影响因子：
4.6
通讯作者：
Yamamoto, Takashi
中科院分区：
文献类型：
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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
转录激活因子样效应子核酸酶（TALEN）是一种位点特异性核酸酶，可以自由设计和容易构建。最近已经报道了构建具有不同TALE支架和重复变体的TALEN的许多方法。然而，尚未比较结构上不同的TALEN的功能。在这里，我们报告了几种类型的TALEN靶向相同位点之间的功能差异。使用基于HEK 293 T细胞的单链退火和Cel-I核酸酶测定，我们发现具有包含非重复可变双残基（非RVD）变异的重复模式化重复变体的TALEN（铂TALEN）显示出比没有非RVD变异的TALEN更高的活性。此外，铂TALENs介导的基因破坏效率在青蛙和大鼠中显着高于以前的报告。因此，这项研究证明了一个有效的系统，用于构建这些高活性的铂TALEN（铂门系统），这可能会建立一个新的标准，在TALEN工程。
Transcription activator-like effector (TALE) nuclease (TALEN) is a site-specific nuclease, which can be freely designed and easily constructed. Numerous methods of constructing TALENs harboring different TALE scaffolds and repeat variants have recently been reported. However, the functionalities of structurally different TALENs have not yet been compared. Here, we report on the functional differences among several types of TALENs targeting the same loci. Using HEK293T cell-based single-strand annealing and Cel-I nuclease assays, we found that TALENs with periodically-patterned repeat variants harboring non-repeat-variable di-residue (non-RVD) variations (Platinum TALENs) showed higher activities than TALENs without non-RVD variations. Furthermore, the efficiencies of gene disruption mediated by Platinum TALENs in frogs and rats were significantly higher than in previous reports. This study therefore demonstrated an efficient system for the construction of these highly active Platinum TALENs (Platinum Gate system), which could establish a new standard in TALEN engineering.
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影响因子：
4.6
作者：
Mashimo, Tomoji;Kaneko, Takehito;Sakuma, Tetsushi;Kobayashi, Junya;Kunihiro, Yayoi;Voigt, Birger;Yamamoto, Takashi;Serikawa, Tadao
通讯作者：
Serikawa, Tadao
影响因子：
46.9
作者：
通讯作者：
--
影响因子：
46.9
作者：
Kim, Yongsub;Kweon, Jiyeon;Kim, Jin-Soo
通讯作者：
Kim, Jin-Soo
影响因子：
--
作者：
Reyon, Deepak;Khayter, Cyd;Regan, Maureen R;Joung, J Keith;Sander, Jeffry D
通讯作者：
Sander, Jeffry D
影响因子：
23.9
作者：
Ding, Qiurong;Lee, Youn-Kyoung;Schaefer, Esperance A. K.;Peters, Derek T.;Veres, Adrian;Kim, Kevin;Kuperwasser, Nicolas;Motola, Daniel L.;Meissner, Torsten B.;Hendriks, William T.;Trevisan, Marta;Gupta, Rajat M.;Moisan, Annie;Banks, Eric;Friesen, Max;Schinzel, Robert T.;Xia, Fang;Tang, Alexander;Xia, Yulei;Figueroa, Emmanuel;Wann, Amy;Ahfeldt, Tim;Daheron, Laurence;Zhang, Feng;Rubin, Lee L.;Peng, Lee F.;Chung, Raymond T.;Musunuru, Kiran;Cowan, Chad A.
通讯作者：
Cowan, Chad A.