PUBLICATION

A Multifunctional Mutagenesis System for Analysis of Gene Function in Zebrafish

Authors
Quach, H.N., Tao, S., Vrljicak, P., Joshi, A., Ruan, H., Sukumaran, R., Varshney, G.K., LaFave, M.C., Screen Team, T.D., Burgess, S.M., Winkler, C., Emelyanov, A., Parinov, S., Sampath, K.
ID
ZDB-PUB-150404-2
Date
2015
Source
G3 (Bethesda)   5(6): 1283-99 (Journal)
Registered Authors
Burgess, Shawn, Joshi, Adita, Parinov, Serguei, Quach, Helen Ngoc Bao, Sampath, Karuna, Sukumaran, Rashmi, Tao, Shijie, Varshney, Gaurav, Vrljicak, Pavle, Winkler, Christoph
Keywords
Ac/Ds transposon, functional genomics, gene expression, insertional mutagenesis
MeSH Terms
  • Animals
  • Base Sequence
  • Chromosome Mapping
  • Enhancer Elements, Genetic
  • Fluorescence
  • Gene Expression Profiling
  • Genes, Reporter
  • Genetic Loci
  • Molecular Sequence Data
  • Mutagenesis, Insertional/methods*
  • Mutation/genetics
  • Organ Specificity/genetics
  • Phenotype
  • Zebrafish/genetics*
  • Zebrafish Proteins/genetics
(all 15)
PubMed
25840430 Full text @ G3 (Bethesda)
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 in zebrafish to facilitate functional genomic studies in this model of human development and disease.
Genes / Markers
Marker Marker Type Name
aglaGENEamylo-alpha-1, 6-glucosidase, 4-alpha-glucanotransferase a
bcl2bGENEBCL2 apoptosis regulator b
cdc42ep3GENECDC42 effector protein (Rho GTPase binding) 3
dhx37GENEDEAH (Asp-Glu-Ala-His) box polypeptide 37
dnah6GENEdynein, axonemal, heavy chain 6
gfapGENEglial fibrillary acidic protein
kdrlGENEkinase insert domain receptor like
msi2aGENEmusashi RNA-binding protein 2a
ndnflGENEneuron-derived neurotrophic factor , like
opa1GENEOPA1 mitochondrial dynamin like GTPase
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Figures
Figure Gallery (5 images)
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Expression
Phenotype
Fish Conditions Stage Phenotype Figure
WT + MO3-dhx37standard conditionsDay 5
WT + MO3-dhx37standard conditionsDay 5
WT + MO3-dhx37standard conditionsDay 5
dhx37ws09771Tg/ws09771Tgstandard conditionsDay 5
dhx37ws09771Tg/ws09771Tgstandard conditionsDay 5
dhx37ws09771Tg/ws09771Tgstandard conditionsDay 5
unm_ws0069tgws0069Tg/ws0069Tgstandard conditionsProtruding-mouth
unm_ws01962tgws01962Tg/ws01962Tgstandard conditionsLong-pec
unm_ws01962tgws01962Tg/ws01962Tgstandard conditionsLong-pec
unm_ws01962tgws01962Tg/ws01962Tgstandard conditionsLong-pec
1 - 10 of 13
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Mutations / Transgenics
Allele Construct Type Affected Genomic Region
ws0028TgTransgenic Insertion
    ws0069TgTransgenic Insertion
    ws0093TgTransgenic Insertion
      ws0148TgTransgenic Insertion
      ws0188TgTransgenic Insertion
        ws0194TgTransgenic Insertion
          ws0308TgTransgenic Insertion
            ws0310TgTransgenic Insertion
              ws0449TgTransgenic Insertion
              ws0504TgTransgenic Insertion
                1 - 10 of 162
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                Human Disease / Model
                No data available
                Sequence Targeting Reagents
                Target Reagent Reagent Type
                dhx37MO3-dhx37MRPHLNO
                dhx37MO4-dhx37MRPHLNO
                1 - 2 of 2
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                Fish
                Fish
                aglaws09772Tg
                cdc42ep3ws2110Tg
                dhx37ws09771Tg
                dhx37ws09771Tg/ws09771Tg
                dnah6ws22931Tg
                msi2aws1208Tg
                ndnflws21321Tg
                opa1ws1357Tg
                ptprubws0148Tg
                pxdnws0449Tg
                1 - 10 of 167
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                Antibodies
                No data available
                Orthology
                Engineered Foreign Genes
                Marker Marker Type Name
                EGFPEFGEGFP
                mCherryEFGmCherry
                1 - 2 of 2
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                Mapping
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                Citation
                Quach, H.N., Tao, S., Vrljicak, P., Joshi, A., Ruan, H., Sukumaran, R., Varshney, G.K., LaFave, M.C., Screen Team, T.D., Burgess, S.M., Winkler, C., Emelyanov, A., Parinov, S., Sampath, K. (2015) A Multifunctional Mutagenesis System for Analysis of Gene Function in Zebrafish. G3 (Bethesda). 5(6):1283-99.