PUBLICATION

Photomorphs: A novel light-activated reagent for controlling gene expression in zebrafish

Authors
Tomasini, A.J., Schuler, A.D., Zebala, J.A., and Mayer, A.N.
ID
ZDB-PUB-090807-4
Date
2009
Source
Genesis (New York, N.Y. : 2000)   47(11): 736-743 (Journal)
Registered Authors
Mayer, Alan, Schuler, Aaron D.
Keywords
morpholino, gene expression, zebrafish, gene knockdown, light activatable, caged morpholino
MeSH Terms
  • Animals
  • Base Sequence
  • Cadherins/genetics
  • DNA Primers
  • Gene Expression Regulation/drug effects*
  • Gene Knockdown Techniques
  • Green Fluorescent Proteins/genetics
  • Indicators and Reagents/pharmacology*
  • Light*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Zebrafish/embryology
  • Zebrafish/genetics*
PubMed
19644983 Full text @ Genesis
Abstract
Manipulating gene expression in zebrafish is critical for exploiting the full potential of this vertebrate model organism. Morpholino oligos are the most commonly used antisense technology for knocking down gene expression. However, morpholinos suffer from a lack of control over the timing and location of knockdown. In this report, we describe a novel light- activatable knockdown reagent called PhotoMorph. PhotoMorphs can be generated from existing morpholinos by hybridization with a complementary caging strand containing a photocleavable linkage. The caging strand neutralizes the morpholino activity until irradiation of the PhotoMorph with UV light releases the morpholino. We generated PhotoMorphs to target genes encoding enhanced green fluorescent protein, No tail, and E-cadherin to illustrate the utility of this approach. Temporal control of gene expression with PhotoMorphs permitted us to circumvent the early lethal phenotype of E-cadherin knockdown. A splice-blocking PhotoMorph directed to the rheb gene showed light-dependent gene knockdown up to 72 hpf. PhotoMorphs thus offer a new class of laboratory reagents suitable for the spatiotemporal control of gene expression in the zebrafish.
Genes / Markers
Figures
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping