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Updated: Jul 17, 2026
Hyperactive piggyBac Transposase-mediated Germline Transformation in the Fall Armyworm, Spodoptera frugiperda
Published on: September 23, 2021
Germ line transformation of the silkworm, Bombyx mori, using the transposable element Minos
K Uchino1, M Imamura, K Shimizu
1Transgenic Silkworm Research Center, National Institute of Agrobiological Sciences, 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan.
Abstract:
We investigated the use of Minos as a vector for transgenesis in the silkworm, Bombyx mori. We first constructed a vector plasmid with the green fluorescent protein (GFP) gene fused with the silkworm cytoplasmic actin gene (A3) promoter, and a helper plasmid with the Minos transposase gene controlled by the same A3 promoter. Injection of the vector and helper plasmid DNA into silkworm eggs produced transgenic animals in the following generation. The efficiency of transgenic silkworm production using this method was much lower than that obtained using piggyBac-mediated germ line transformation. However, >40-fold increase in the efficiency of producing transgenic silkworms was obtained using an in vitro synthesized source of Minos transposase mRNA. We conclude that the Minos transposon is a useful vector for construction of transgenic silkworms, particularly when in vitro synthesized mRNA is used. This is the first report showing that Minos can be used as a vector for germ-line transformation in lepidopteran insects.
Insights
Minos transposon enables germ-line transformation in silkworms. Using Minos transposase mRNA significantly boosts transgenic silkworm production efficiency, offering a new tool for lepidopteran insect research.
Area of Science:
- * Entomology
- * Molecular Biology
- * Genetic Engineering
Background:
- * Transgenic silkworms are crucial for research and industry.
- * Existing methods like piggyBac have limitations.
- * Novel transposon systems are needed for efficient gene transfer.
Purpose of the Study:
- * To evaluate Minos as a transgenesis vector in Bombyx mori (silkworm).
- * To optimize Minos-mediated transformation efficiency.
- * To establish Minos for germ-line transformation in lepidopteran insects.
Main Methods:
- * Construction of Minos vector and helper plasmids with GFP and transposase genes.
- * Injection of plasmid DNA into silkworm eggs.
- * Comparison of efficiency with piggyBac system.
- * Utilization of in vitro synthesized Minos transposase mRNA.
Main Results:
- * Transgenic silkworms were produced using Minos vector and helper plasmids.
- * Initial efficiency was lower than piggyBac-mediated transformation.
- * In vitro synthesized Minos transposase mRNA increased efficiency by over 40-fold.
- * Minos demonstrated potential for germ-line transformation in lepidopterans.
Conclusions:
- * Minos transposon is a viable vector for silkworm transgenesis.
- * In vitro synthesized Minos transposase mRNA significantly enhances transformation efficiency.
- * This study represents the first use of Minos for germ-line transformation in lepidopteran insects.
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