1] National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Chaoyang District, Beijing, China. [2] School of Life Sciences, Peking University, Beijing, China. [3].
Nat Biotechnol. 2013 Oct;31(10):934-7. doi: 10.1038/nbt.2674. Epub 2013 Aug 18.
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.
我们开发了一种通过在体细胞中表达转录激活因子样效应物核酸酶(TALEN)来产生条件性基因敲除的方法。利用通过诱导或组织特异性启动子控制的质粒进行种系转化,我们在特定的发育阶段和组织中观察到了有效的基因修饰和表型。我们进一步利用这种方法绕过了 cor-1 的胚胎要求,cor-1 编码人类严重联合免疫缺陷(SCID)蛋白 coronin 的同源物,我们确定了它在幼虫 Q 细胞谱系中的细胞迁移中的重要作用。我们的结果表明,在模式生物的体细胞中表达的 TALEN 为功能基因组学提供了一种通用工具。