Establishment of an integrated automated embryonic manipulation system for producing genetically modified mice

Abstract Genetically modified mice are commonly used in biologic, medical, and drug discovery research, but conventional microinjection methods used for genetic modification require extensive training and practical experience. Here we present a fully automated system for microinjection into the pron...

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Autores principales:	Tomoo Eto, Hiroki Ueda, Ryoji Ito, Tsukasa Takahashi, Toshiaki Watanabe, Motohito Goto, Yusuke Sotomaru, Nobuaki Tanaka, Riichi Takahashi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6be20c5eef20463a9492fcfe4c2710ab
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spelling	oai:doaj.org-article:6be20c5eef20463a9492fcfe4c2710ab2021-12-02T15:02:32ZEstablishment of an integrated automated embryonic manipulation system for producing genetically modified mice10.1038/s41598-021-91148-92045-2322https://doaj.org/article/6be20c5eef20463a9492fcfe4c2710ab2021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91148-9https://doaj.org/toc/2045-2322Abstract Genetically modified mice are commonly used in biologic, medical, and drug discovery research, but conventional microinjection methods used for genetic modification require extensive training and practical experience. Here we present a fully automated system for microinjection into the pronucleus to facilitate genetic modification. We first developed software that automatically controls the microinjection system hardware. The software permits automatic rotation of the zygote to move the pronucleus to the injection pipette insertion position. We also developed software that recognizes the pronucleus in 3-dimensional coordinates so that the injection pipette can be automatically inserted into the pronucleus, and achieved a 94% insertion rate by linking the 2 pieces of software. Next, we determined the optimal solution injection conditions (30 hPa, 0.8–2.0 s) by examining the survival rate of injected zygotes. Finally, we produced transgenic (traditional DNA injection and piggyBac Transposon system) and knock-in (genomic editing) mice using our newly developed Integrated Automated Embryo Manipulation System (IAEMS). We propose that the IAEMS will simplify highly reproducible pronuclear stage zygote microinjection procedures.Tomoo EtoHiroki UedaRyoji ItoTsukasa TakahashiToshiaki WatanabeMotohito GotoYusuke SotomaruNobuaki TanakaRiichi TakahashiNature 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 Tomoo Eto Hiroki Ueda Ryoji Ito Tsukasa Takahashi Toshiaki Watanabe Motohito Goto Yusuke Sotomaru Nobuaki Tanaka Riichi Takahashi Establishment of an integrated automated embryonic manipulation system for producing genetically modified mice
description	Abstract Genetically modified mice are commonly used in biologic, medical, and drug discovery research, but conventional microinjection methods used for genetic modification require extensive training and practical experience. Here we present a fully automated system for microinjection into the pronucleus to facilitate genetic modification. We first developed software that automatically controls the microinjection system hardware. The software permits automatic rotation of the zygote to move the pronucleus to the injection pipette insertion position. We also developed software that recognizes the pronucleus in 3-dimensional coordinates so that the injection pipette can be automatically inserted into the pronucleus, and achieved a 94% insertion rate by linking the 2 pieces of software. Next, we determined the optimal solution injection conditions (30 hPa, 0.8–2.0 s) by examining the survival rate of injected zygotes. Finally, we produced transgenic (traditional DNA injection and piggyBac Transposon system) and knock-in (genomic editing) mice using our newly developed Integrated Automated Embryo Manipulation System (IAEMS). We propose that the IAEMS will simplify highly reproducible pronuclear stage zygote microinjection procedures.
format	article
author	Tomoo Eto Hiroki Ueda Ryoji Ito Tsukasa Takahashi Toshiaki Watanabe Motohito Goto Yusuke Sotomaru Nobuaki Tanaka Riichi Takahashi
author_facet	Tomoo Eto Hiroki Ueda Ryoji Ito Tsukasa Takahashi Toshiaki Watanabe Motohito Goto Yusuke Sotomaru Nobuaki Tanaka Riichi Takahashi
author_sort	Tomoo Eto
title	Establishment of an integrated automated embryonic manipulation system for producing genetically modified mice
title_short	Establishment of an integrated automated embryonic manipulation system for producing genetically modified mice
title_full	Establishment of an integrated automated embryonic manipulation system for producing genetically modified mice
title_fullStr	Establishment of an integrated automated embryonic manipulation system for producing genetically modified mice
title_full_unstemmed	Establishment of an integrated automated embryonic manipulation system for producing genetically modified mice
title_sort	establishment of an integrated automated embryonic manipulation system for producing genetically modified mice
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/6be20c5eef20463a9492fcfe4c2710ab
work_keys_str_mv	AT tomooeto establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice AT hirokiueda establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice AT ryojiito establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice AT tsukasatakahashi establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice AT toshiakiwatanabe establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice AT motohitogoto establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice AT yusukesotomaru establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice AT nobuakitanaka establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice AT riichitakahashi establishmentofanintegratedautomatedembryonicmanipulationsystemforproducinggeneticallymodifiedmice
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Establishment of an integrated automated embryonic manipulation system for producing genetically modified mice

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