CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.

The water flea Daphnia magna has been used as an animal model in ecology, evolution, and environmental sciences. Thanks to the recent progress in Daphnia genomics, genetic information such as the draft genome sequence and expressed sequence tags (ESTs) is now available. To investigate the relationsh...

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Autores principales:	Takashi Nakanishi, Yasuhiko Kato, Tomoaki Matsuura, Hajime Watanabe
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0d0c4dff4ce94de7a0951b9c5f6a4aaf
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spelling	oai:doaj.org-article:0d0c4dff4ce94de7a0951b9c5f6a4aaf2021-11-18T08:17:50ZCRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.1932-620310.1371/journal.pone.0098363https://doaj.org/article/0d0c4dff4ce94de7a0951b9c5f6a4aaf2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24878568/?tool=EBIhttps://doaj.org/toc/1932-6203The water flea Daphnia magna has been used as an animal model in ecology, evolution, and environmental sciences. Thanks to the recent progress in Daphnia genomics, genetic information such as the draft genome sequence and expressed sequence tags (ESTs) is now available. To investigate the relationship between phenotypes and the available genetic information about Daphnia, some gene manipulation methods have been developed. However, a technique to induce targeted mutagenesis into Daphnia genome remains elusive. To overcome this problem, we focused on an emerging genome editing technique mediated by the clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR/Cas) system to introduce genomic mutations. In this study, we targeted a functionally conserved regulator of eye development, the eyeless gene in D. magna. When we injected Cas9 mRNAs and eyeless-targeting guide RNAs into eggs, 18-47% of the survived juveniles exhibited abnormal eye morphology. After maturation, up to 8.2% of the adults produced progenies with deformed eyes, which carried mutations in the eyeless loci. These results showed that CRISPR/Cas system could introduce heritable mutations into the endogenous eyeless gene in D. magna. This is the first report of a targeted gene knockout technique in Daphnia and will be useful in uncovering Daphnia gene functions.Takashi NakanishiYasuhiko KatoTomoaki MatsuuraHajime WatanabePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 5, p e98363 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Takashi Nakanishi Yasuhiko Kato Tomoaki Matsuura Hajime Watanabe CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.
description	The water flea Daphnia magna has been used as an animal model in ecology, evolution, and environmental sciences. Thanks to the recent progress in Daphnia genomics, genetic information such as the draft genome sequence and expressed sequence tags (ESTs) is now available. To investigate the relationship between phenotypes and the available genetic information about Daphnia, some gene manipulation methods have been developed. However, a technique to induce targeted mutagenesis into Daphnia genome remains elusive. To overcome this problem, we focused on an emerging genome editing technique mediated by the clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR/Cas) system to introduce genomic mutations. In this study, we targeted a functionally conserved regulator of eye development, the eyeless gene in D. magna. When we injected Cas9 mRNAs and eyeless-targeting guide RNAs into eggs, 18-47% of the survived juveniles exhibited abnormal eye morphology. After maturation, up to 8.2% of the adults produced progenies with deformed eyes, which carried mutations in the eyeless loci. These results showed that CRISPR/Cas system could introduce heritable mutations into the endogenous eyeless gene in D. magna. This is the first report of a targeted gene knockout technique in Daphnia and will be useful in uncovering Daphnia gene functions.
format	article
author	Takashi Nakanishi Yasuhiko Kato Tomoaki Matsuura Hajime Watanabe
author_facet	Takashi Nakanishi Yasuhiko Kato Tomoaki Matsuura Hajime Watanabe
author_sort	Takashi Nakanishi
title	CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.
title_short	CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.
title_full	CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.
title_fullStr	CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.
title_full_unstemmed	CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.
title_sort	crispr/cas-mediated targeted mutagenesis in daphnia magna.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/0d0c4dff4ce94de7a0951b9c5f6a4aaf
work_keys_str_mv	AT takashinakanishi crisprcasmediatedtargetedmutagenesisindaphniamagna AT yasuhikokato crisprcasmediatedtargetedmutagenesisindaphniamagna AT tomoakimatsuura crisprcasmediatedtargetedmutagenesisindaphniamagna AT hajimewatanabe crisprcasmediatedtargetedmutagenesisindaphniamagna
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CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.

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