PUBLICATION

Intersectional Gene Expression in Zebrafish Using the Split KalTA4 System

Authors
Almeida, R.G., Lyons, D.A.
ID
ZDB-PUB-151021-2
Date
2015
Source
Zebrafish   12(6): 377-86 (Journal)
Registered Authors
Almeida, Rafael, Lyons, David A.
Keywords
none
MeSH Terms
  • Promoter Regions, Genetic
  • Zebrafish/genetics
  • Zebrafish/metabolism*
  • Genes, Reporter
  • Animals, Genetically Modified
  • Green Fluorescent Proteins
  • DNA-Binding Proteins/genetics
  • DNA-Binding Proteins/metabolism*
  • Transcription Factors/genetics
  • Transcription Factors/metabolism*
  • Animals
  • Gene Expression Regulation/physiology*
  • Zebrafish Proteins/genetics
  • Zebrafish Proteins/metabolism*
  • Trans-Activators/metabolism
  • SOXE Transcription Factors/genetics
  • SOXE Transcription Factors/metabolism
  • Protein Structure, Tertiary
  • Embryo, Nonmammalian/metabolism
  • Plasmids
(all 20)
PubMed
26485616 Full text @ Zebrafish
Abstract
In this study, we describe the adaptation of the split Gal4 system for zebrafish. The Gal4-UAS system is widely used for expression of genes-of-interest by crossing driver lines expressing the transcription factor Gal4 (under the control of the promoter of interest) with reporter lines where upstream activating sequence (UAS) repeats (recognized by Gal4) drive expression of the genes-of-interest. In the Split Gal4 system, hemi-drivers separately encode the DNA-binding domain (DBD) and the activation domain (AD) of Gal4. When encoded under two different promoters, only those cells in the intersection of the promoters' expression pattern and in which both promoters are active reconstitute a functional Gal4 and activate expression from a UAS-driven transgene. We split the zebrafish-optimized version of Gal4, KalTA4, and generated a hemi-driver encoding the KalTA4 DBD and a hemi-driver encoding KalTA4's AD. We show that split KalTA4 domains can assemble in vivo and transactivate a UAS reporter transgene and that each hemi-driver alone cannot transactivate the reporter. Also, transactivation can happen in several cell types, with similar efficiency to intact KalTA4. Finally, in transient mosaic expression assays, we show that when hemi-drivers are preceded by two distinct promoters, they restrict the expression of an UAS-driven reporter from a broader pattern (sox10) to its constituent smaller neuronal pattern. The Split KalTA4 system should be useful for expression of genes-of-interest in an intersectional manner, allowing for more refined manipulations of cell populations in zebrafish.
Genes / Markers
Figures
Figure Gallery (3 images)
Show all Figures
Expression
Phenotype
No data available
Mutations / Transgenics
Allele Construct Type Affected Genomic Region
s1999tTgTransgenic Insertion
    ue1TgTransgenic Insertion
      ue7TgTransgenic Insertion
        vu12TgTransgenic Insertion
          vu234TgTransgenic Insertion
            1 - 5 of 5
            Show
            Human Disease / Model
            No data available
            Sequence Targeting Reagents
            No data available
            Fish
            No data available
            Antibodies
            No data available
            Orthology
            No data available
            Engineered Foreign Genes
            Marker Marker Type Name
            EGFPEFGEGFP
            KaedeEFGKaede
            KALTA4EFGKALTA4
            mRFPEFGmRFP
            1 - 4 of 4
            Show
            Mapping
            No data available
            Your Input Welcome
            We welcome your input and comments. Please use this form to recommend updates to the information in ZFIN. We appreciate as much detail as possible and references as appropriate. We will review your comments promptly.
            Citation
            Almeida, R.G., Lyons, D.A. (2015) Intersectional Gene Expression in Zebrafish Using the Split KalTA4 System. Zebrafish. 12(6):377-86.