CRISPR/Cas9-mediated rapid generation of multiple mouse lines identified Ccdc63 as essential for spermiogenesis

Young, Samantha A.M.; Miyata, Haruhiko; Satouh, Yuhkoh; Kato, Hirotaka; Nozawa, Kaori; Isotani, Ayako; Aitken, R. John; Baker, Mark A.; Ikawa, Masahito

doi:https://doi.org/10.3390/ijms161024732

CRISPR/Cas9-mediated rapid generation of multiple mouse lines identified Ccdc63 as essential for spermiogenesis

Young, Samantha A.M.; Miyata, Haruhiko; Satouh, Yuhkoh et al.

International Journal of Molecular Sciences, 2015, 16(10), 24732-24750

アクセス数:207件（2025-04-18 03:11 集計）

固定URL: https://hdl.handle.net/11094/78595

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タイトル	CRISPR/Cas9-mediated rapid generation of multiple mouse lines identified Ccdc63 as essential for spermiogenesis
著者	著者名 Young, Samantha A.M.
	NRID 1000050604732 1000050604732 researchmap permalink hmiyata hmiyata Scopus Author ID 37161837000 37161837000 ORCID 0000-0003-4758-5803 0000-0003-4758-5803 著者名 Miyata, Haruhiko
	NRID 1000040545571 1000040545571 著者名 Satouh, Yuhkoh
	著者名 Kato, Hirotaka
	著者名 Nozawa, Kaori
	NRID 1000020444523 1000020444523 著者名 Isotani, Ayako
	著者名 Aitken, R. John
	著者名 Baker, Mark A.
	NRID 1000020304066 1000020304066 ORCID 0000-0001-9859-6217 0000-0001-9859-6217 researchmap permalink read0054910 read0054910 Scopus Author ID 16689054500 16689054500 著者名 Ikawa, Masahito
キーワード等	Genome editing
	Sperm motility
	Spermatogenesis
	Targeted mutagenesis
内容	Young, S.A.M.; Miyata, H.; Satouh, Y.; Kato, H.; Nozawa, K.; Isotani, A.; Aitken, R.J.; Baker, M.A.; Ikawa, M. CRISPR/Cas9-Mediated Rapid Generation of Multiple Mouse Lines Identified Ccdc63 as Essential for Spermiogenesis. Int. J. Mol. Sci. 2015, 16, 24732-24750. https://doi.org/10.3390/ijms161024732
抄録	Spermatozoa are flagellated cells whose role in fertilization is dependent on their ability to move towards an oocyte. The structure of the sperm flagella is highly conserved across species, and much of what is known about this structure is derived from studies utilizing animal models. One group of proteins essential for the movement of the flagella are the dyneins. Using the advanced technology of CRISPR/Cas9 we have targeted three dynein group members; Dnaic1, Wdr63 and Ccdc63 in mice. All three of these genes are expressed strongly in the testis. We generated mice with amino acid substitutions in Dnaic1 to analyze two specific phosphorylation events at S124 and S127, and generated simple knockouts of Wdr63 and Ccdc63. We found that the targeted phosphorylation sites in Dnaic1 were not essential for male fertility. Similarly, Wdr63 was not essential for male fertility; however, Ccdc63 removal resulted in sterile male mice due to shortened flagella. This study demonstrates the versatility of the CRISPR/Cas9 system to generate animal models of a highly complex system by introducing point mutations and simple knockouts in a fast and efficient manner.
出版者	MDPI
収録物名	International Journal of Molecular Sciences
巻(号)	16(10)
ページ	24732-24750
刊行年月	2015-10-16
言語	英語
ハンドルURL	https://hdl.handle.net/11094/78595
PISSN	16616596
eISSN	14220067
NCID	AA12038549
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.3390/ijms161024732
関連情報 (isIdenticalTo)	PMID 26501274
アクセス権	open access
権利情報	© 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCTERMS.bibliographicCitation	International Journal of Molecular Sciences. 2015, 16(10), p. 24732-24750
DC.title	CRISPR/Cas9-mediated rapid generation of multiple mouse lines identified Ccdc63 as essential for spermiogenesis
DC.creator	Young, Samantha A.M.
	Miyata, Haruhiko
	Satouh, Yuhkoh
	Kato, Hirotaka
	Nozawa, Kaori
	Isotani, Ayako
	Aitken, R. John
	Baker, Mark A.
	Ikawa, Masahito
DC.publisher	MDPI
DC.language' scheme='DCTERMS.RFC1766	英語
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2015-10-16
DC.identifier	https://doi.org/10.3390/ijms161024732
DC.subject	Genome editing
	Sperm motility
	Spermatogenesis
	Targeted mutagenesis
DC.description	Young, S.A.M.; Miyata, H.; Satouh, Y.; Kato, H.; Nozawa, K.; Isotani, A.; Aitken, R.J.; Baker, M.A.; Ikawa, M. CRISPR/Cas9-Mediated Rapid Generation of Multiple Mouse Lines Identified Ccdc63 as Essential for Spermiogenesis. Int. J. Mol. Sci. 2015, 16, 24732-24750. https://doi.org/10.3390/ijms161024732
DCTERMS.abstract	Spermatozoa are flagellated cells whose role in fertilization is dependent on their ability to move towards an oocyte. The structure of the sperm flagella is highly conserved across species, and much of what is known about this structure is derived from studies utilizing animal models. One group of proteins essential for the movement of the flagella are the dyneins. Using the advanced technology of CRISPR/Cas9 we have targeted three dynein group members; Dnaic1, Wdr63 and Ccdc63 in mice. All three of these genes are expressed strongly in the testis. We generated mice with amino acid substitutions in Dnaic1 to analyze two specific phosphorylation events at S124 and S127, and generated simple knockouts of Wdr63 and Ccdc63. We found that the targeted phosphorylation sites in Dnaic1 were not essential for male fertility. Similarly, Wdr63 was not essential for male fertility; however, Ccdc63 removal resulted in sterile male mice due to shortened flagella. This study demonstrates the versatility of the CRISPR/Cas9 system to generate animal models of a highly complex system by introducing point mutations and simple knockouts in a fast and efficient manner.
DC.rights	© 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	78595
citation_title	CRISPR/Cas9-mediated rapid generation of multiple mouse lines identified Ccdc63 as essential for spermiogenesis
citation_author	Young, Samantha A.M.
	Miyata, Haruhiko
	Satouh, Yuhkoh
	Kato, Hirotaka
	Nozawa, Kaori
	Isotani, Ayako
	Aitken, R. John
	Baker, Mark A.
	Ikawa, Masahito
citation_publisher	MDPI
citation_language	英語
citation_date	2015-10-16
citation_journal_title	International Journal of Molecular Sciences
citation_volume	16
citation_issue	10
citation_firstpage	24732
citation_lastpage	24750
citation_doi	https://doi.org/10.3390/ijms161024732
citation_pmid	26501274
citation_public_url	https://hdl.handle.net/11094/78595

CRISPR/Cas9-mediated rapid generation of multiple mouse lines identified Ccdc63 as essential for spermiogenesis

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