title: Quantitatively relating gene expression to light intensity via the serial connection of blue light sensor and CRISPRi.
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.
authors.affiliation: Bio-X Institutes, Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders (Ministry of Education), ‡2013 SJTU-BioX-Shanghai Team for International Genetically Engineered Machine (iGEM) Competition, Shanghai Jiao Tong University , Shanghai 200240, China.
authors.forename: Hongyi
authors.initials: H
authors.lastname: Wu
authors.forename: Yushu
authors.initials: Y
authors.lastname: Wang
authors.forename: You
authors.initials: Y
authors.lastname: Wang
authors.forename: Xinang
authors.initials: X
authors.lastname: Cao
authors.forename: Yifan
authors.initials: Y
authors.lastname: Wu
authors.forename: Zhuofei
authors.initials: Z
authors.lastname: Meng
authors.forename: Qiang
authors.initials: Q
authors.lastname: Su
authors.forename: Zhongying
authors.initials: Z
authors.lastname: Wang
authors.forename: Shuai
authors.initials: S
authors.lastname: Yang
authors.forename: Weijian
authors.initials: W
authors.lastname: Xu
authors.forename: Shiyi
authors.initials: S
authors.lastname: Liu
authors.forename: Pan
authors.initials: P
authors.lastname: Cheng
authors.forename: Jianxuan
authors.initials: J
authors.lastname: Wu
authors.forename: Md Rezaul Islam
authors.initials: MR
authors.lastname: Khan
authors.forename: Lin
authors.initials: L
authors.lastname: He
authors.forename: Gang
authors.initials: G
authors.lastname: Ma
mesh_terms: D057005:Bioengineering / Q000379:methods*; D064113:CRISPR-Cas Systems / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000235:genetics* / Q000528:radiation effects*; D008027:Light; D010957:Plasmids / Q000235:genetics; D034622:RNA Interference