Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish

Abstract Direct tests of gene function have historically been performed in a limited number of model organisms. The CRISPR/Cas system is species-agnostic, offering the ability to manipulate genes in a range of models, enabling insights into evolution, development, and physiology. Astatotilapia burto...

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Autores principales:	Cheng-Yu Li, Joshua R. Steighner, Garrett Sweatt, Tod R. Thiele, Scott A. Juntti
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c07cdcf014ce43579c59e8a7131330b5
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spelling	oai:doaj.org-article:c07cdcf014ce43579c59e8a7131330b52021-12-02T16:26:22ZManipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish10.1038/s41598-021-94577-82045-2322https://doaj.org/article/c07cdcf014ce43579c59e8a7131330b52021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-94577-8https://doaj.org/toc/2045-2322Abstract Direct tests of gene function have historically been performed in a limited number of model organisms. The CRISPR/Cas system is species-agnostic, offering the ability to manipulate genes in a range of models, enabling insights into evolution, development, and physiology. Astatotilapia burtoni, a cichlid fish from the rivers and shoreline around Lake Tanganyika, has been extensively studied in the laboratory to understand evolution and the neural control of behavior. Here we develop protocols for the creation of CRISPR-edited cichlids and create a broadly useful mutant line. By manipulating the Tyrosinase gene, which is necessary for eumelanin pigment production, we describe a fast and reliable approach to quantify and optimize gene editing efficiency. Tyrosinase mutants also remove a major obstruction to imaging, enabling visualization of subdermal structures and fluorophores in situ. These protocols will facilitate broad application of CRISPR/Cas9 to studies of cichlids as well as other non-traditional model aquatic species.Cheng-Yu LiJoshua R. SteighnerGarrett SweattTod R. ThieleScott A. JunttiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Cheng-Yu Li Joshua R. Steighner Garrett Sweatt Tod R. Thiele Scott A. Juntti Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish
description	Abstract Direct tests of gene function have historically been performed in a limited number of model organisms. The CRISPR/Cas system is species-agnostic, offering the ability to manipulate genes in a range of models, enabling insights into evolution, development, and physiology. Astatotilapia burtoni, a cichlid fish from the rivers and shoreline around Lake Tanganyika, has been extensively studied in the laboratory to understand evolution and the neural control of behavior. Here we develop protocols for the creation of CRISPR-edited cichlids and create a broadly useful mutant line. By manipulating the Tyrosinase gene, which is necessary for eumelanin pigment production, we describe a fast and reliable approach to quantify and optimize gene editing efficiency. Tyrosinase mutants also remove a major obstruction to imaging, enabling visualization of subdermal structures and fluorophores in situ. These protocols will facilitate broad application of CRISPR/Cas9 to studies of cichlids as well as other non-traditional model aquatic species.
format	article
author	Cheng-Yu Li Joshua R. Steighner Garrett Sweatt Tod R. Thiele Scott A. Juntti
author_facet	Cheng-Yu Li Joshua R. Steighner Garrett Sweatt Tod R. Thiele Scott A. Juntti
author_sort	Cheng-Yu Li
title	Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish
title_short	Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish
title_full	Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish
title_fullStr	Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish
title_full_unstemmed	Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish
title_sort	manipulation of the tyrosinase gene permits improved crispr/cas editing and neural imaging in cichlid fish
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/c07cdcf014ce43579c59e8a7131330b5
work_keys_str_mv	AT chengyuli manipulationofthetyrosinasegenepermitsimprovedcrisprcaseditingandneuralimagingincichlidfish AT joshuarsteighner manipulationofthetyrosinasegenepermitsimprovedcrisprcaseditingandneuralimagingincichlidfish AT garrettsweatt manipulationofthetyrosinasegenepermitsimprovedcrisprcaseditingandneuralimagingincichlidfish AT todrthiele manipulationofthetyrosinasegenepermitsimprovedcrisprcaseditingandneuralimagingincichlidfish AT scottajuntti manipulationofthetyrosinasegenepermitsimprovedcrisprcaseditingandneuralimagingincichlidfish
_version_	1718384078335508480

Manipulation of the Tyrosinase gene permits improved CRISPR/Cas editing and neural imaging in cichlid fish

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