TIGHTLY CONTROLLED TETRACYCLINE-INDUCIBLE TRANSCRIPTION SYSTEM FOR EXPLOSIVE GENE EXPRESSION IN CULTURED SILKWORM CELLS

Authors

  • Tsuneyuki Tatsuke,

    1. Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki, Fukuoka, Japan
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  • Jae Man Lee,

    1. Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki, Fukuoka, Japan
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  • Takahiro Kusakabe,

    Corresponding author
    • Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki, Fukuoka, Japan
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  • Kazuhiro Iiyama,

    1. Laboratory of Insect Pathology and Microbial Control, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, Hakozaki, Fukuoka, Japan
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  • Hideki Sezutsu,

    1. Transgenic Silkworm Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan
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  • Keiro Uchino

    1. Transgenic Silkworm Research Unit, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, Japan
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  • Present address of T. Tatsuke: Laboratory for Bioinformatics, Graduate School of Systems Life Sciences, Kyushu University, 3–1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.

  • Grant sponsor: KAKENHI; Grant number: 22248003; Grant sponsor: Japan Society for the Promotion of Science; Grant number: 23580077.

Correspondence to: T. Kusakabe, Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu University, 6-10-1 Hakozaki, Fukuoka 812-8581, Japan.

E-mail: kusakabe@agr.kyushu-u.ac.jp

Abstract

Tetracycline-inducible gene expression (Tet-on) system is a particularly powerful tool for transgenic research and has become one of the first choices for the control of transgene expression in a mammal and a fly. Previously, we have generated the modified reverse tetracycline-controlled transactivators and tetO promoters for a Bombyx mori Tet-on system. In order to further improve this system, Giant, a transcriptional silencer from Drosophila melanogaster, is introduced to repress leaky transcription in the absence of doxycycline. Further, the promoter responsibility to the tetracycline-controlled transactivators is facilitated by introducing a synthetic minimal core promoter. With the tightly regulated second-generation silkworm Tet-on system, we obtain up to 300-fold induction of gene expression with the addition of doxycycline.

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