TAEL: a zebrafish-optimized optogenetic gene expression system with fine spatial and temporal control

Here, we describe an optogenetic gene expression system optimized for use in zebrafish. This system overcomes the limitations of current inducible expression systems by enabling robust spatial and temporal regulation of gene expression in living organisms. Because existing optogenetic systems show t...

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Bibliographic Details
Published inDevelopment (Cambridge) Vol. 144; no. 2; pp. 345 - 355
Main Authors Reade, Anna, Motta-Mena, Laura B, Gardner, Kevin H, Stainier, Didier Y, Weiner, Orion D, Woo, Stephanie
Format Journal Article
LanguageEnglish
Published England The Company of Biologists Ltd 15.01.2017
Subjects
Animals
Animals, Genetically Modified
Calibration
CRISPR
CRISPR-Cas Systems - genetics
Danio rerio
Developmental stages
Ectoderm
Embryo, Nonmammalian
Embryogenesis
Gene expression
Gene Expression Regulation, Developmental - radiation effects
Genes, Reporter - radiation effects
Light
Optogenetics - methods
Optogenetics - standards
Signal Transduction - genetics
Signal Transduction - radiation effects
Techniques and Resources
Toxicity
Zebrafish
Zebrafish - embryology
Zebrafish - genetics
CRISPR
Nodal
Zebrafish
Optogenetics
Cas9
Gene expression
Endoderm
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Summary:Here, we describe an optogenetic gene expression system optimized for use in zebrafish. This system overcomes the limitations of current inducible expression systems by enabling robust spatial and temporal regulation of gene expression in living organisms. Because existing optogenetic systems show toxicity in zebrafish, we re-engineered the blue-light-activated EL222 system for minimal toxicity while exhibiting a large range of induction, fine spatial precision and rapid kinetics. We validate several strategies to spatially restrict illumination and thus gene induction with our new TAEL (TA4-EL222) system. As a functional example, we show that TAEL is able to induce ectopic endodermal cells in the presumptive ectoderm via targeted sox32 induction. We also demonstrate that TAEL can be used to resolve multiple roles of Nodal signaling at different stages of embryonic development. Finally, we show how inducible gene editing can be achieved by combining the TAEL and CRISPR/Cas9 systems. This toolkit should be a broadly useful resource for the fish community.
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Present address: School of Natural Sciences, University of California-Merced, Merced, CA 95343, USA.
Present address: Optologix, Inc., 1910 Pacific Ave., Suit 20000, Dallas, TX 75201, USA.
ISSN:0950-1991
1477-9129
DOI:10.1242/dev.139238