Conditional targeted genome editing using somatically expressed TALELENs in C. elegans
Conditional targeted genome editing using somatically expressed TALELENs in C. elegans
复制标题
使用线虫体细胞表达的 TALELEN 进行条件靶向基因组编辑
DOI：
10.1038/nbt.2674
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发表时间：
2013-10-01
影响因子：
46.9
通讯作者：
Ou, Guangshuo
中科院分区：
文献类型：
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作者：
Cheng, Ze;Yi, Peishan;Ou, Guangshuo
我们已经开发了一种在秀丽隐杆线虫中通过在体细胞中表达转录激活因子样效应物核酸酶（TALEN）来产生条件性敲除的方法。使用在诱导型或组织特异性启动子的控制下编码TALEN的质粒进行种系转化，我们观察到有效的基因修饰和在特定发育阶段和组织中产生的表型。我们进一步使用这种方法来绕过胚胎要求的cor-1，它编码的同源物的人类严重联合免疫缺陷（SCID）蛋白冠状蛋白，我们确定了其在幼虫Q细胞系的细胞迁移的重要作用。我们的研究结果表明，在模式生物体细胞中表达的TALEN为功能基因组学提供了一个通用的工具。
We have developed a method for the generation of conditional knockouts in Caenorhabditis elegans by expressing transcription activator-like effector nucleases (TALENs) in somatic cells. Using germline transformation with plasmids encoding TALENs under the control of an inducible or tissue-specific promoter, we observed effective gene modifications and resulting phenotypes in specific developmental stages and tissues. We further used this method to bypass the embryonic requirement of cor-1, which encodes the homolog of human severe combined immunodeficiency (SCID) protein coronin, and we determined its essential role in cell migration in larval Q-cell lineages. Our results show that TALENs expressed in the somatic cells of model organisms provide a versatile tool for functional genomics.