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Title: A hyperactive piggyBac transposon system is an easy-to-implement method for introducing foreign genes into mouse preimplantation embryos.
Authors: Suzuki, Shinnosuke
Tsukiyama, Tomoyuki
Kaneko, Takehito  kyouindb  KAKEN_id
Imai, Hiroshi
Minami, Naojiro  kyouindb  KAKEN_id
Author's alias: 南, 直治郎
Keywords: Cytoplasmic injection
Hyperactive piggyBac transposase
Transgenic mice
Issue Date: 2015
Publisher: Society for Reproduction and Development
Journal title: The Journal of reproduction and development
Volume: 61
Issue: 3
Start page: 241
End page: 244
Abstract: Transgenic mice are important tools for genetic analysis. A current prominent method for producing transgenic mice involves pronuclear microinjection into 1-cell embryos. However, the total transgenic efficiency obtained using this method is less than 10%. Here, we demonstrate that highly efficient transgenesis in mice can be achieved by cytoplasmic microinjection using a hyperactive piggyBac system. In embryos in which hyPBase mRNA and pPB-CAG-TagRFP DNA were co-injected into the cytoplasm, TagRFP fluorescence was observed after the 2-cell stage; when 30 ng/µl pPB-CAG-TagRFP DNA and 30 ng/µl hyPBase mRNA were co-injected, 94.4% of blastocysts were TagRFP positive. Furthermore, a high concentration of hyPBase mRNA resulted in creation of mosaic embryos in which the TagRFP signals partially disappeared. However, suitable concentrations of injected DNA and hyPBase mRNA produced embryos in which almost all blastomeres were TagRFP positive. Thus, the hyperactive piggyBac transposon system is an easy-to-implement and highly effective method that can contribute to production of transgenic mice.
Rights: © 2015 Society for Reproduction and Development
This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License<http://creativecommons.org/licenses/by-nc-nd/3.0/>
URI: http://hdl.handle.net/2433/201842
DOI(Published Version): 10.1262/jrd.2014-157
PubMed ID: 25740401
Appears in Collections:Journal Articles

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