» Articles » PMID: 27993986

TAEL: a Zebrafish-optimized Optogenetic Gene Expression System with Fine Spatial and Temporal Control

Overview
Journal Development
Specialty Biology
Date 2016 Dec 21
PMID 27993986
Citations 39
Authors
Affiliations
Soon will be listed here.
Abstract

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.

Citing Articles

Optogenetic control of phosphate-responsive genes using single component fusion proteins in .

Cleere M, Gardner K bioRxiv. 2024; .

PMID: 39131330 PMC: 11312615. DOI: 10.1101/2024.08.02.605841.


The N-Terminal Region of the BcWCL1 Photoreceptor Is Necessary for Self-Dimerization and Transcriptional Activation upon Light Stimulation in Yeast.

Guerrero M, Ruiz C, Romero A, Robeson L, Ruiz D, Salinas F Int J Mol Sci. 2023; 24(15).

PMID: 37569251 PMC: 10418492. DOI: 10.3390/ijms241511874.


Multidimensional characterization of inducible promoters and a highly light-sensitive LOV-transcription factor.

Gligorovski V, Sadeghi A, Rahi S Nat Commun. 2023; 14(1):3810.

PMID: 37369667 PMC: 10300134. DOI: 10.1038/s41467-023-38959-8.


Spatial and temporal control of expression with light-gated LOV-LexA.

Ribeiro I, Essbauer W, Kutlesa R, Borst A G3 (Bethesda). 2022; 12(10).

PMID: 35876796 PMC: 9526042. DOI: 10.1093/g3journal/jkac178.


A Single-Component Blue Light-Induced System Based on EL222 in .

Wang Z, Yan Y, Zhang H Int J Mol Sci. 2022; 23(11).

PMID: 35683022 PMC: 9181742. DOI: 10.3390/ijms23116344.


References
1.
Kimmel C, Warga R, Schilling T . Origin and organization of the zebrafish fate map. Development. 1990; 108(4):581-94. DOI: 10.1242/dev.108.4.581. View

2.
Kurita R, Sagara H, Aoki Y, Link B, Arai K, Watanabe S . Suppression of lens growth by alphaA-crystallin promoter-driven expression of diphtheria toxin results in disruption of retinal cell organization in zebrafish. Dev Biol. 2003; 255(1):113-27. DOI: 10.1016/s0012-1606(02)00079-9. View

3.
Knopf F, Schnabel K, Haase C, Pfeifer K, Anastassiadis K, Weidinger G . Dually inducible TetON systems for tissue-specific conditional gene expression in zebrafish. Proc Natl Acad Sci U S A. 2010; 107(46):19933-8. PMC: 2993373. DOI: 10.1073/pnas.1007799107. View

4.
Polstein L, Gersbach C . Light-inducible spatiotemporal control of gene activation by customizable zinc finger transcription factors. J Am Chem Soc. 2012; 134(40):16480-3. PMC: 3468123. DOI: 10.1021/ja3065667. View

5.
Crosson S, Rajagopal S, Moffat K . The LOV domain family: photoresponsive signaling modules coupled to diverse output domains. Biochemistry. 2003; 42(1):2-10. DOI: 10.1021/bi026978l. View