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| Transgenic Efficiency of the Cytoplasmic Microinjection Technique Mediated with Different Transposons |
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Abstract In order to investigate the transgenic efficiency of cytoplasm microinject mediated with SB (sleeping beauty), PB (piggyBac) and Tol2 transposons in mice (Mus musculus) and zebrafish (Danio rerio); We coinjected the plasmid of recombinant vector pT3-PST-CAG-GFP containing 3 transposons and the plasmids of SB, PB and Tol2 transposes expression vectors into the cytoplasm of mice and zebrafish zygote with a ratio of 1∶1 respectively, and detected the positive expression ratio of green fluorescent protein (GFP) by fluorescence microscope after cultivation in vitro. The results revealed that the highest transgenic efficiency among 3 transposons was PB (N=281), which reached 52% in 4 Ed (96 h) (Ed: embryo day) mice embryos, significantly higher than (P＜0.05) SB (47%, N=328) and Tol2 (36%, N=273). In the stage of 36 and 60 hpf (hour past fertilization) zebrafish embryos, the highest transgenic efficiency was Tol2, and the GFP expression rate was 36%(N=677) and 37%(N=685), respectively, significant higher than PB at 36 hpf (30%, N=691) and 60 hpf (32%, N=708), and SB at 36 hpf (28%, N=687) and 60 hpf (27%, N=675) respectively; The results indicated that high transgenic efficiency was achieved by using the cytoplasm microinjection technique mediated with transposons, and the transposition activity of different transposons displayed species difference, the highest gene transfer efficiency was PB transposon in mammal embryos, while in fish embryos, Tol2 transposon was the most efficient. This study provides basic reference for the improvement of animal transgenic efficiency.
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Received: 15 May 2015
Published: 23 November 2015
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