title: A Multifunctional Mutagenesis System for Analysis of Gene Function in Zebrafish.
abstract: Since the sequencing of the human reference genome, many human disease-related genes have been discovered. However, understanding the functions of all the genes in the genome remains a challenge. The biological activities of these genes are usually investigated in model organisms such as mice and zebrafish. Large-scale mutagenesis screens to generate disruptive mutations are useful for identifying and understanding the activities of genes. Here, we report a multifunctional mutagenesis system in zebrafish using the maize Ds transposon. Integration of the Ds transposable element containing an mCherry reporter for protein trap events and an EGFP reporter for enhancer trap events produced a collection of transgenic lines marking distinct cell and tissue types, and mutagenized genes in the zebrafish genome by trapping and prematurely terminating endogenous protein coding sequences. We obtained 642 zebrafish lines with dynamic reporter gene expression. The characterized fish lines with specific expression patterns will be made available through the European Zebrafish Resource Center (EZRC), and a database of reporter expression is available online (http://fishtrap.warwick.ac.uk/). Our approach complements other efforts using zebrafish to facilitate functional genomic studies in this model of human development and disease.
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.
authors.forename: Helen Ngoc Bao
authors.initials: HN
authors.lastname: Quach
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604 Department of Biological Sciences, National University of Singapore, Singapore 117543.
authors.forename: Shijie
authors.initials: S
authors.lastname: Tao
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604 Division of Biomedical Cell Biology, Warwick Medical School, Coventry, United Kingdom CV4 7AJ.
authors.forename: Pavle
authors.initials: P
authors.lastname: Vrljicak
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.
authors.forename: Adita
authors.initials: A
authors.lastname: Joshi
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.
authors.forename: Hua
authors.initials: H
authors.lastname: Ruan
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.
authors.forename: Rashmi
authors.initials: R
authors.lastname: Sukumaran
authors.affiliation: National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892-8004.
authors.forename: Gaurav K
authors.initials: GK
authors.lastname: Varshney
authors.affiliation: National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892-8004.
authors.forename: Matthew C
authors.initials: MC
authors.lastname: LaFave
authors.affiliation: National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892-8004.
authors.forename: Shawn M
authors.initials: SM
authors.lastname: Burgess
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543 Centre for Bioimaging Sciences, National University of Singapore, Singapore 117543.
authors.forename: Christoph
authors.initials: C
authors.lastname: Winkler
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.
authors.forename: Alexander
authors.initials: A
authors.lastname: Emelyanov
authors.affiliation: Temasek Life Sciences Laboratory, National University of Singapore, Singapore 117604.
authors.forename: Sergey
authors.initials: S
authors.lastname: Parinov
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543 Division of Biomedical Cell Biology, Warwick Medical School, Coventry, United Kingdom CV4 7AJ K.Sampath@warwick.ac.uk.
authors.forename: Karuna
authors.initials: K
authors.lastname: Sampath
mesh_terms: D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D004742:Enhancer Elements, Genetic; D005453:Fluorescence; D020869:Gene Expression Profiling; D017930:Genes, Reporter; D056426:Genetic Loci; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods*; D009154:Mutation / Q000235:genetics; D009928:Organ Specificity / Q000235:genetics; D010641:Phenotype; D015027:Zebrafish / Q000235:genetics*; D029961:Zebrafish Proteins / Q000235:genetics
chemical_list: D029961:Zebrafish Proteins
keywords: Ac/Ds transposon; functional genomics; gene expression; insertional mutagenesis
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