Nihongaki Yuta, Yamamoto Shun, Kawano Fuun, Suzuki Hideyuki, Sato Moritoshi
Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.
Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.
Chem Biol. 2015 Feb 19;22(2):169-74. doi: 10.1016/j.chembiol.2014.12.011. Epub 2015 Jan 22.
Targeted endogenous gene activation is necessary for understanding complex gene networks and has great potential in medical and industrial applications. The CRISPR-Cas system offers simple and powerful tools for this purpose. However, these CRISPR-Cas-based tools for activating user-defined genes are unable to offer precise temporal control of gene expression, despite the fact that many biological phenomena are regulated by highly dynamic patterns of gene expression. Here we created a light-inducible, user-defined, endogenous gene activation system based on CRISPR-Cas9. We demonstrated that this CRISPR-Cas9-based transcription system can allow rapid and reversible targeted gene activation by light. In addition, using this system, we have exemplified photoactivation of multiple user-defined endogenous genes in mammalian cells. The present CRISPR-Cas9-based transcription system offers simple and versatile approaches for precise endogenous gene activation in basic biological research and biotechnology applications.
靶向内源性基因激活对于理解复杂的基因网络至关重要,并且在医学和工业应用中具有巨大潜力。CRISPR-Cas系统为此提供了简单而强大的工具。然而,尽管许多生物学现象受基因表达的高度动态模式调控,但这些基于CRISPR-Cas的用于激活用户定义基因的工具无法实现对基因表达的精确时间控制。在此,我们基于CRISPR-Cas9创建了一种光诱导、用户定义的内源性基因激活系统。我们证明,这种基于CRISPR-Cas9的转录系统能够通过光实现快速且可逆的靶向基因激活。此外,利用该系统,我们在哺乳动物细胞中例证了多个用户定义的内源性基因的光激活。当前基于CRISPR-Cas9的转录系统为基础生物学研究和生物技术应用中的精确内源性基因激活提供了简单且通用的方法。