A hyperactive piggyBac transposon system is an easy-to-implement method for introducing foreign genes into mouse preimplantation embryos.
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.
Laboratory of Reproductive Biology, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502, Japan.
Shinnosuke
S
Suzuki
Tomoyuki
T
Tsukiyama
Takehito
T
Kaneko
Hiroshi
H
Imai
Naojiro
N
Minami
D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004251:DNA Transposable Elements; D046149:Embryo Culture Techniques / Q000379:methods; D005260:Female; D018014:Gene Transfer Techniques; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D008845:Microinjections; D012333:RNA, Messenger / Q000378:metabolism; D019076:Transgenes; D000097573:Red Fluorescent Protein
D004251:DNA Transposable Elements; D008164:Luminescent Proteins; D012333:RNA, Messenger; D000097573:Red Fluorescent Protein