Quantitatively Relating Gene Expression to Light Intensity via the Serial Connection of Blue Light Sensor and CRISPRi

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• 作者：Hongyi Wu ; Yushu Wang ; You Wang ; Xinang Cao ; Yifan Wu ; Zhuofei Meng ; Qiang Su ; Zhongying Wang ; Shuai Yang ; Weijian Xu ; Shiyi Liu ; Pan Cheng ; Jianxuan Wu ; Md. Rezaul Islam Khan ; Lin He ; Gang Ma
• 刊名：ACS Synthetic Biology
• 出版年：2014
• 出版时间：December 19, 2014
• 年：2014
• 卷：3
• 期：12
• 页码：979-982
• 全文大小：208K
• ISSN：2161-5063

文摘

The ability to regulate endogenous gene expression is critical in biological research. Existing technologies, such as RNA interference, zinc-finger regulators, transcription-activator-like effectors, and CRISPR-mediated regulation, though proved to be competent in significantly altering expression levels, do not provide a quantitative adjustment of regulation effect. As a solution to this problem, we place CRISPR-mediated interference under the control of blue light: while dCas9 protein is constitutively expressed, guide RNA transcription is regulated by YF1-FixJ-P_FixK2, a blue light responding system. With a computer-controlled luminous device, the quantitative relationship between target gene expression and light intensity has been determined. As the light intensifies, the expression level of target gene gradually ascends. This remarkable property enables sensor-CRISPRi to accurately interrogate cellular activities.