title: Enhancer activity-based identification of functional enhancers using zebrafish embryos.
abstract: Chromatin immunoprecipitation (ChIP) against enhancer-associated marks with massive sequencing is a powerful approach to identify genome-wide distributions of putative enhancers. However, functional in vivo analysis is required to elucidate the activities of predicted enhancers. Using zebrafish embryos, we established a ChIP-Injection method that enables identification of functional enhancers based on their enhancer activities in embryos. Each reporter gene possessing the enhancer-associated genomic region enriched by ChIP was injected into zebrafish embryos to analyze the activity of putative enhancers. By using the ChIP-Injection, we identified 32 distinct putative enhancers that drove specific expression. Additionally, we generated transgenic lines that exhibit distributions of the EGFP signal as was observed in the screening. Furthermore, the expression pattern driven by the identified somite-specific enhancer resembled that of the gene acta2. The results indicate that ChIP-Injection provides an efficient approach for identification of active enhancers in a potentially wide variety of developmental tissues and stages.
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Tomohito
authors.initials: T
authors.lastname: Taminato
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Yokota
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Soh
authors.initials: S
authors.lastname: Araki
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Ovara
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan.
authors.forename: Kyo
authors.initials: K
authors.lastname: Yamasu
authors.affiliation: Division of Life Science, Graduate School of Science and Engineering, Saitama University, Shimo-Okubo, Sakura-ku, Saitama City, Saitama 338-8570, Japan. Electronic address: akawamur@mail.saitama-u.ac.jp.
authors.forename: Akinori
authors.initials: A
authors.lastname: Kawamura
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D047369:Chromatin Immunoprecipitation / Q000379:methods*; D004742:Enhancer Elements, Genetic; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D023281:Genomics; D049452:Green Fluorescent Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Cis-regulatory element; Enhancer; Tailbud; Zebrafish