A Powerful CRISPR/Cas9-Based Method for Targeted Transcriptional Activation
A Powerful CRISPR/Cas9-Based Method for Targeted Transcriptional Activation
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DOI：
10.1002/anie.201601708
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发表时间：
2016-05-23
影响因子：
16.6
通讯作者：
Kondo, Toru
中科院分区：
文献类型：
--
作者：
Katayama, Shota;Moriguchi, Tetsuo;Kondo, Toru
内源性基因的靶向转录激活对于理解生理转录网络、合成遗传回路和诱导细胞表型变化是重要的。CRISPR/Cas9系统具有实现这一目的的巨大潜力，然而，它尚未成功地用于有效激活内源性基因并诱导细胞表型的变化。开发了一种通过使用CRISPR/Cas9进行转录激活的强大方法。通过CRISPR/Cas9用未甲基化的启动子替换甲基化的启动子足以激活HEK 293 T细胞中神经细胞基因OLIG 2和胚胎干细胞基因NANOG的表达。此外，基于CRISPR/Cas9的OLIG 2活化诱导胚胎癌细胞系NTERA-2表达神经元标记物β III-微管蛋白。
Targeted transcriptional activation of endogenous genes is important for understanding physiological transcriptional networks, synthesizing genetic circuits, and inducing cellular phenotype changes. The CRISPR/Cas9 system has great potential to achieve this purpose, however, it has not yet been successfully used to efficiently activate endogenous genes and induce changes in cellular phenotype. A powerful method for transcriptional activation by using CRISPR/Cas9 was developed. Replacement of a methylated promoter with an unmethylated one by CRISPR/Cas9 was sufficient to activate the expression of the neural cell gene OLIG2 and the embryonic stem cell gene NANOG in HEK293T cells. Moreover, CRISPR/Cas9-based OLIG2 activation induced the embryonic carcinoma cell line NTERA-2 to express the neuronal marker beta III-tubulin.