1. Quantitatively relating gene expression to light intensity via the serial connection of blue light sensor and CRISPRi
- Author
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Zhuofei Meng, Shiyi Liu, Hongyi Wu, Jianxuan Wu, Shuai Yang, You Wang, Xinang Cao, Weijian Xu, Yushu Wang, Yifan Wu, Zhongying Wang, Pan Cheng, Qiang Su, Lin He, Gang Ma, and Rezaul Islam Khan
- Subjects
Light ,Effector ,Biomedical Engineering ,Gene Expression ,Endogeny ,Bioengineering ,General Medicine ,Biology ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Molecular biology ,Cell biology ,Light intensity ,Transcription (biology) ,RNA interference ,Gene expression ,Escherichia coli ,RNA Interference ,Guide RNA ,CRISPR-Cas Systems ,Blue light ,Plasmids - Abstract
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-PFixK2, 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.
- Published
- 2014