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

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...

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Bibliographic Details
Published in:Scientific reports 2021-06, Vol.11 (1), p.11770-11770, Article 11770
Main Authors: Eto, Tomoo, Ueda, Hiroki, Ito, Ryoji, Takahashi, Tsukasa, Watanabe, Toshiaki, Goto, Motohito, Sotomaru, Yusuke, Tanaka, Nobuaki, Takahashi, Riichi
Format: Article
Language:English
Subjects:
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Automation
Computer programs
Embryos
Humanities and Social Sciences
Injection
Insertion
Microinjection
multidisciplinary
Pronucleus
Science
Science (multidisciplinary)
Software
Survival
Zygotes
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Summary: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-021-91148-9