title: Fluorescence-Activated Cell Sorting and Gene Expression Profiling of GFP-Positive Cells from Transgenic Zebrafish Lines.
abstract: Gene expression profiling is a useful approach for deeper understanding of the specificity of cells, tissues, and organs in the transcriptional level. Recent development of high-throughput next-generation sequence (NGS) allows the RNA-seq method for this profiling. This method provides precise information of transcripts about the quantitation and the structure such as the splicing variants. In this chapter, we describe a method for gene expression profiling of GFP-positive cells from transgenic zebrafish by RNA-seq. We labeled specific cells in the brain with GFP by crossing a Gal4 driver line with the UAS:GFP line, isolated those cells by fluorescence-activated cell sorting (FACS), and analyzed by RNA-seq.
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizouka, 411-8540, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Tanabe
authors.affiliation: Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Chiba, 277-8562, Japan.
authors.forename: Masahide
authors.initials: M
authors.lastname: Seki
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizouka, 411-8540, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Itoh
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizouka, 411-8540, Japan.|Department of Genetics, Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizouka, 411-8540, Japan.
authors.forename: Ailani
authors.initials: A
authors.lastname: Deepak
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizouka, 411-8540, Japan.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Lal
authors.affiliation: Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Chiba, 277-8562, Japan.
authors.forename: Terumi
authors.initials: T
authors.lastname: Horiuchi
authors.affiliation: Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, Chiba, 277-8562, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Suzuki
authors.affiliation: Division of Molecular and Developmental Biology, National Institute of Genetics, Mishima, Shizouka, 411-8540, Japan. kokawaka@nig.ac.jp.|Department of Genetics, Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizouka, 411-8540, Japan. kokawaka@nig.ac.jp.
authors.forename: Koichi
authors.initials: K
authors.lastname: Kawakami
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D059014:High-Throughput Nucleotide Sequencing; D015027:Zebrafish / Q000235:genetics / Q000378:metabolism*
chemical_list: D049452:Green Fluorescent Proteins
keywords: Enhancer trap; FACS; GFP; Gal4-UAS system; Gene expression analysis; Gene trap; Next-generation sequence; RNA extract; RNA-seq; Tol2 transgenesis