Bacterial delivery of TALEN proteins for human genome editing.
Bacterial delivery of TALEN proteins for human genome editing.
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用于人类基因组编辑的 TALEN 蛋白的细菌传递
DOI：
10.1371/journal.pone.0091547
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Jin S
中科院分区：
文献类型：
--
作者：
Jia J;Jin Y;Bian T;Wu D;Yang L;Terada N;Wu W;Jin S
转录激活因子样效应物核酸酶（TALEN）是一类新型的序列特异性核酸酶，其最近因其易于生产和在基因组编辑中的高效性而变得突出。TALEN对识别特定的DNA序列，并在靶位点引入双链断裂，引发非同源末端连接和同源重组。目前的TALEN递送方法涉及通过转染引入外源遗传物质，如质粒DNA或mRNA。在这里，我们展示了TALEN递送的另一种方式，细菌III型分泌系统（T3SS）介导的TALEN蛋白直接注射到人细胞中。细菌注射的TALEN显示出有效靶向宿主细胞核，在那里它持续几乎12小时。使用靶向venus基因的一对TALEN，这种注射的核TALEN显示出在靶位点中引入DNA突变的功能。有趣的是，S期细胞似乎对TALEN介导的靶基因修饰表现出更大的敏感性。因此，这种基因组编辑的效率可以容易地通过感染剂量、重复感染的次数以及S期细胞的富集来操纵。这项工作进一步扩展了T3SS在将功能蛋白质递送到哺乳动物细胞中以改变其生物医学应用特征方面的效用。
Transcription Activator-Like Effector Nucleases (TALENs) are a novel class of sequence-specific nucleases that have recently gained prominence for its ease of production and high efficiency in genome editing. A TALEN pair recognizes specific DNA sequences and introduce double-strand break in the target site, triggering non-homologous end joining and homologous recombination. Current methods of TALEN delivery involves introduction of foreign genetic materials, such as plasmid DNA or mRNA, through transfection. Here, we show an alternative way of TALEN delivery, bacterial type III secretion system (T3SS) mediated direct injection of the TALEN proteins into human cells. Bacterially injected TALEN was shown to efficiently target host cell nucleus where it persists for almost 12 hours. Using a pair of TALENs targeting venus gene, such injected nuclear TALENs were shown functional in introducing DNA mutation in the target site. Interestingly, S-phase cells seem to show greater sensitivity to the TALEN mediated target gene modification. Accordingly, efficiency of such genome editing can easily be manipulated by the infection dose, number of repeated infections as well as enrichment of S phase cells. This work further extends the utility of T3SS in the delivery of functional proteins into mammalian cells to alter their characters for biomedical applications.
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影响因子：
46.9
作者：
Hockemeyer, Dirk;Soldner, Frank;Beard, Caroline;Gao, Qing;Mitalipova, Maisam;DeKelver, Russell C.;Katibah, George E.;Amora, Ranier;Boydston, Elizabeth A.;Zeitler, Bryan;Meng, Xiangdong;Miller, Jeffrey C.;Zhang, Lei;Rebar, Edward J.;Gregory, Philip D.;Urnov, Fyodor D.;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf
影响因子：
3.6
作者：
Büttner, D;Gürlebeck, D;Bonas, U
通讯作者：
Bonas, U
DOI：
10.1073/pnas.1019533108
发表时间：
2011-02-08
影响因子：
11.1
作者：
Mahfouz, Magdy M.;Li, Lixin;Zhu, Jian-Kang
通讯作者：
Zhu, Jian-Kang
影响因子：
5.8
作者：
Grau, Jan;Boch, Jens;Posch, Stefan
通讯作者：
Posch, Stefan
影响因子：
2.1
作者：
Pastor, A;Chabert, J;Attree, I
通讯作者：
Attree, I