Construction of artificial promoters sensitively responsive to sonication in vitro

  • Akihiko Watanabe
  • Satoshi Kakutani
  • Ryohei OgawaEmail author
  • Sung-il Lee
  • Toru Yoshida
  • Akihiro Morii
  • Go Kagiya
  • Loreto B. FerilJr.
  • Hideki Fuse
  • Takashi Kondo
Original Article



To develop artificial promoters that are activated in response to sonication and to determine these properties in vitro.


The binding sites of four transcription factors (nuclear factor-kappa B, activating protein-1, nuclear factor-Y, and CArG element binding factor A) that are activated by oxidative stress were randomly ligated and linked to a TATA-box sequence to control the luciferase gene located downstream. Transiently transfected HeLa cells from human cervical cancer with a plasmid vector containing such a gene cassette were exposed to sonication, and enhancement of luciferase expression was assessed by dual luciferase assay.


Of 62 promoters constructed, two promoters, designated clone 31 and clone 62 promoters, showed a more than tenfold enhancement 6 h after sonication with 1-MHz ultrasound at 1.0 W/cm2 for 60 s. These promoters were activated in a dose-dependent manner with the intensity and duration of sonication. The activation was attenuated by addition of dimethyl sulfoxide, an antioxidant, suggesting that oxidative stress was involved. The clone 31 promoter responded to each of two serial sonications. When sonicated 24 h after the first sonication, the peak of promoter enhancement was higher than that after the first sonication.


A promoter sensitively responsive to sonication was constructed using the above method, possibly leading to the construction of a promoter of interest that could be applied for clinical use.


promoter transcription oxidative stress 


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Copyright information

© The Japan Society of Ultrasonics in Medicine 2009

Authors and Affiliations

  • Akihiko Watanabe
    • 1
  • Satoshi Kakutani
    • 2
  • Ryohei Ogawa
    • 2
    Email author
  • Sung-il Lee
    • 3
  • Toru Yoshida
    • 4
  • Akihiro Morii
    • 1
  • Go Kagiya
    • 5
  • Loreto B. FerilJr.
    • 6
  • Hideki Fuse
    • 1
  • Takashi Kondo
    • 2
  1. 1.Department of Urology, Graduate School of Medicine and Pharmaceutical SciencesUniversity of ToyamaToyamaJapan
  2. 2.Department of Radiological Sciences, Graduate School of Medicine and Pharmaceutical SciencesUniversity of ToyamaToyamaJapan
  3. 3.Institute of Biomedical ScienceKansai Medical UniversityMoriguchiJapan
  4. 4.Second Department of Surgery, Graduate School of Medicine and Pharmaceutical SciencesUniversity of ToyamaToyamaJapan
  5. 5.School of Allied Health SciencesKitasato UniversityKanagawaJapan
  6. 6.Department of Anatomy, School of MedicineFukuoka UniversityFukuokaJapan

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