title: CRISPR-Cas9-based photoactivatable transcription system.
abstract: 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.
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Nihongaki
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.
authors.forename: Shun
authors.initials: S
authors.lastname: Yamamoto
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.
authors.forename: Fuun
authors.initials: F
authors.lastname: Kawano
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Suzuki
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan. Electronic address: cmsato@mail.ecc.u-tokyo.ac.jp.
authors.forename: Moritoshi
authors.initials: M
authors.lastname: Sato
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D019556:COS Cells; D064113:CRISPR-Cas Systems; D002522:Chlorocebus aethiops; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008027:Light; D008163:Luminescent Measurements; D011401:Promoter Regions, Genetic; D061785:Transcription Initiation, Genetic
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; C065500:ASCL1 protein, human