Conditional knockdown of target gene expression by tetracycline regulated transcription of double strand RNA

Authors

  • Xubin Hou,

    1. Department of Molecular Neurobiology, Graduate School of Life Sciences/Institute of Development, Aging and Cancer, Tohoku University, Seiryo-machi 4-1, Aoba-ku, Sendai 980-8575
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    • Present address: Laboratory of Neuronal Development Department of Medical and Dental Sciences, Niigata University, 1-757 Asahimachi Chuo-ku Niigata 951-8510 Japan.

    • Hou was a Japanese Government (Monbukagakusho) Scholarship Student.

  • Minoru Omi,

    1. Department of Molecular Neurobiology, Graduate School of Life Sciences/Institute of Development, Aging and Cancer, Tohoku University, Seiryo-machi 4-1, Aoba-ku, Sendai 980-8575
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  • Hidekiyo Harada,

    1. Department of Molecular Neurobiology, Graduate School of Life Sciences/Institute of Development, Aging and Cancer, Tohoku University, Seiryo-machi 4-1, Aoba-ku, Sendai 980-8575
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    • Present address: Division of Cell & Developmental Biology, College of Life Sciences, University of Dundee, Dow Street, DD1 5EH, UK.

  • Shunsuke Ishii,

    1. Laboratory of Molecular Genetics, RIKEN Tsukuba Institute, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074
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  • Yoshiko Takahashi,

    1. Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara 630-0192, Japan
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  • Harukazu Nakamura

    Corresponding author
    1. Department of Molecular Neurobiology, Graduate School of Life Sciences/Institute of Development, Aging and Cancer, Tohoku University, Seiryo-machi 4-1, Aoba-ku, Sendai 980-8575
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Author to whom all correspondence should be addressed.
Email: nakamura@idac.tohoku.ac.jp

Abstract

In vivo electroporation has served as an effective tool for the study of developmental biology. Here we report tetracycline inducible gene knockdown by electroporation. Our system consists of genome integration of a cassette encoding long double strand RNA (dsRNA) of a gene of interest by electroporation, transcription of which is assured by RNA polymerase II, and induction of transcription of dsRNA by tetracyclin. Long dsRNA decapped by ribozyme in the cassette and without poly A tail is processed into siRNA within nuclei. We could successfully induce knockdown of En2 and Coactosin by Dox administration.

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