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

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 locat...

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Bibliographic Details
Published in:Genesis (New York, N.Y. : 2000) N.Y. : 2000), 2009-11, Vol.47 (11), p.736-743
Main Authors: Tomasini, Amber J., Schuler, Aaron D., Zebala, John A., Mayer, Alan N.
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Cadherins - genetics
caged morpholino
DNA Primers
gene expression
Gene Expression Regulation - drug effects
gene knockdown
Gene Knockdown Techniques
Green Fluorescent Proteins - genetics
Indicators and Reagents - pharmacology
Light
light activatable
morpholino
Reverse Transcriptase Polymerase Chain Reaction
zebrafish
Zebrafish - embryology
Zebrafish - genetics
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Summary: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. genesis 47:736–743, 2009. © 2009 Wiley‐Liss, Inc.
ISSN:1526-954X
1526-968X
DOI:10.1002/dvg.20554