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Development of real-time intracellular pH sensor using a membrane-permeable peptide nucleic acid.

Research Project

Project/Area Number 16500200
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Neuroscience in general
Research InstitutionOkayama University

Principal Investigator

TOMIZAWA Kazuhito  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Associate Prof., 大学院・医歯薬学総合研究科, 助教授 (40274287)

Co-Investigator(Kenkyū-buntansha) MATSUSHITA Masayuki  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Assistant Prof., 大学院・医歯薬学総合研究科, 講師 (30273965)
MATSUI Hideki  Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (30157234)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2005: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2004: ¥2,300,000 (Direct Cost: ¥2,300,000)
KeywordsPeptide nucleic acid / Polyarginine / Nucleic acid / Bio-molecule / Bio-imaging / Cell / 蛋白質導入法 / TAT / センサー / リアルタイム / イメージング / 蛋白導入法 / FRET
Research Abstract

Membrane-permeable peptide nucleic acid (PNA) developed by us is thought to be a powerful tool for the regulation of cell function. The purpose of this project is that development of the tools that are useful for the regulation of protein expression and the function and for the monitoring of nano-ecology in living cells using the PNA. In the present project, we show that membrane-permeable PNA is a powerful tool for the regulation of gene expression and for anti-cancer therapy as follows.
1.Regulation of gene expression.
We synthesized a PNA, which effectively hybridized with plasmid DNA, and the PNA was fused a membrane-permeable poly-arginine to have the ability of membrane permeability. After mixing the PNA with plasmid DNAs, cells were incubated with the complex and the efficiency of the transfection was investigated. The plasmid DNAs were effectively transfected in both cells and the efficiency is better than that of cationic transfection method.
2.Antigene therapy using membrane-permeable PNA.
We synthesized PNAs that are able to bind with p53 mRNA and the double-strand DNA. Cancer cells were incubated with the PNAs. The PNAs were delivered into cells and inhibited the expression of p53 mRNA and the protein. The combination treatment of the PNAs with p53 protein therapy was capable to replace endogenous mutated p53 proteins with wild-type p53 proteins in the cancer cells.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (28 results)

All 2006 2005 2004

All Journal Article (28 results)

  • [Journal Article] p53 protein transduction therapy : successful targeting and inhibition of the growth of the bladder cancer cells.2006

    • Author(s)
      Inoue, M., Tomizawa, K.et al.
    • Journal Title

      Eur. Urol. 49 ・ 1

      Pages: 161-168

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] p53 protein transduction therapy : Successful targeting and inhibition of the growth of the bladder cancer cells.2006

    • Author(s)
      Inoue, M., Tomizawa, K., Matsushita, M., Lu, Y.-F., Yokoyama, T., Yanai, H., Takashima, A., Kumon, H., Matsui, H.
    • Journal Title

      Eur.Urol. 49・1

      Pages: 161-168

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] The NH-2-terminal of influenza virus hemagglutinin-2 subunit peptides enhance the anti-tumor potency of poly-arginine-mediated p53 protein transduction.2005

    • Author(s)
      Michiue, H., Tomizawa, K.et al.
    • Journal Title

      J. Bio1. Chem. 280 ・ 9

      Pages: 8285-8289

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Ubiquitination-resistant p53 protein transduction therapy facilitates anti-cancer effect on the growth of human malignant glioma cells.2005

    • Author(s)
      Michiue, H., Tomizawa, K.et al.
    • Journal Title

      FEBS Lett. 579 ・ 18

      Pages: 3965-3969

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Cdk5-dependent regulation of glucose-stimulated insulin secretion.2005

    • Author(s)
      Wei, F.-Y., Tomizawa, K.et al.
    • Journal Title

      Nature Med. 11 ・ 10

      Pages: 1104-1108

    • NAID

      130005447816

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Control of cyclin-dependent kinase 5 (Cdk5) activity by glutamatergic regulation of p35 stability.2005

    • Author(s)
      Wei, F.-Y., Tomizawa, K.et al.
    • Journal Title

      J. Neurochem. 93 ・ 2

      Pages: 502-512

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Truncation and activation of calcineurin A by calpain I in Alzheimer disease brain.2005

    • Author(s)
      Liu, F., Tomizawa, K.et a1.
    • Journal Title

      J. Bio1. Chem. 280 ・ 45

      Pages: 37755-37762

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] The NH-2-terminal of influenza virus hemagglutinin-2 subunit peptides enhance the anti-tumor potency of poly-arginine-mediated p53 protein transduction.2005

    • Author(s)
      Michiue, H., Tomizawa, K., Wei, F.-Y., Matsushita, M., Lu, Y.-F., Ichikawa, T., Tamiya, T., Date, I., Matsui, H.
    • Journal Title

      J.Biol.Chem. 280・9

      Pages: 8285-8289

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Ubiquitination-resistant p53 protein transduction therapy facilitates anti-cancer effect on the growth of human malignant glioma cells.2005

    • Author(s)
      Michiue, H., Tomizawa, K., Matsushita, M., Tamiya, T., Lu, Y.-F., Ichikawa, T., Date, I., Matsui, H.
    • Journal Title

      FEBS Lett. 579・18

      Pages: 3965-3969

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Cdk5-dependent regulation of glucose-stimulated insulin secretion.2005

    • Author(s)
      Wei, F.-Y., Nagashima, K., Ohshima, T., Saheki, Y., Lu, Y.-F., Matsushita, M., Yamada, Y., Mikoshiba, K., Seino, Y., Matsui, H., Tomizawa, K.
    • Journal Title

      Nature Med. 11・10

      Pages: 1104-1108

    • NAID

      130005447816

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Control of Cyclin-dependent kinase 5 (Cdk5) activity by glutamatergic regulation of p35 stability.2005

    • Author(s)
      Wei, F.-Y., Tomizawa, K., Ohshima, T., Asada, A., Saito, T., Nguyen C., Bibb, JA., Ishiguro, K., Kulkami, A.B., Pant, H.C., Mikoshiba, K., Matsui, H., Hisanaga, S.
    • Journal Title

      J.Neurochem. 93・2

      Pages: 502-512

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Truncation and activation of calcineurin A by calpain I in Alzheimer disease brain.2005

    • Author(s)
      Liu, F., Grundke-Iqbal, I., Iqbal, K., Oda, Y., Tomizawa, K., Gong C.-X.
    • Journal Title

      J.Biol.Chem. 280・45

      Pages: 37755-37762

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] The NH2-terminal of influenza virus hemagglutinin-2 subunit peptides enhance the anti-tumor potency of poly-arginine-mediated p53 protein transduction.2005

    • Author(s)
      Michiue H, Tomizawa K, Wei FY, Matsushita M, Lu YF, Ichikawa T, Tamiya T, Date I, Matsui H.
    • Journal Title

      J.Biol.Chem. 280(9)

      Pages: 8285-8289

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Calpain inhibitors prevent neuronal cell death and ameliorate motor disturbances after compression-induced spinal cord injury in rats.2005

    • Author(s)
      Arataki S, Tomizawa K, Moriwaki A, Nishida K, Matsushita M, Ozaki T, Kunisada T, Yoshida A, Inoue H, Matsui H.
    • Journal Title

      J.Neurotrauma 22(3)

      Pages: 398-406

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Regulation of synaptic vesicle recycling by calcineurin in different vesicle pools.2005

    • Author(s)
      Kumashiro S, Lu YF, tomizawa K, Matsushita M, Wei FY, Matsui H.
    • Journal Title

      Nerosci.Res. 51(4)

      Pages: 435-443

    • NAID

      130005447570

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Channel function is dissociated from the intrinsic kinase acitivty and autophosphorylation of TRPM7/ChaK1.2005

    • Author(s)
      Matsushita M, Kozak JA, Shimizu Y, McLachlin DT, Yamaguchi H, Wei FY, Tomizawa K, Matsui H, Chait BT, Cahalan MD, Nairn AC.
    • Journal Title

      J.Biol.Chem. 280(21)

      Pages: 20793-20803

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Regulation of N-methyl-D-asparate receptors by calpain in cortical neurons.2005

    • Author(s)
      Wu HY, Yuen EY, Lu YF, Matsushita M, Matsui H, Yan Z, Tomizawa K
    • Journal Title

      J.Biol.Chem. 280(22)

      Pages: 21588-21593

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Ubiquitination-resistant p53 protein transduction therapy facilitates anti-cancer effect on the growth of human malignant glioma cells.2005

    • Author(s)
      Michiue H, Tomizawa K, Matsushita M, Tamiya T, Lu YF, Ichikawa T, Date I, Matsui H.
    • Journal Title

      FEBS Lett. 579(18)

      Pages: 3965-3969

    • Related Report
      2005 Annual Research Report
  • [Journal Article] A new approach to inhibiting astrocytic IP3-induced intracellular calcium increase in an astrocyte-neuron co-culture system.2005

    • Author(s)
      Saheki Y, Li ST, Matsushita M, Wu YM, Cai WH, Wei FY, Lu YF, Moriwaki A, tomizawa K, Matsui H.
    • Journal Title

      Brain Res. 1055(1-2)

      Pages: 196-201

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Cdk5-dependent regulation of glucose-stimulated insulin secretion.2005

    • Author(s)
      Wei FY, Nagashima K, Ohshima T, Saheki Y, Lu YF, Matsushita M, Yamada Y, Mikoshiba K, Seino Y, Matsui H, Tomizawa K.
    • Journal Title

      Nat.Med. 11(10)

      Pages: 1104-1108

    • NAID

      130005447816

    • Related Report
      2005 Annual Research Report
  • [Journal Article] p53 protein transduction therapy : successful targeting and inhibition of the growth of the bladder cancer cells.2005

    • Author(s)
      Inoue M, Tomizawa K, Matsushita M, Lu YF, Yokoyama T, Yanai H, Takashima A, Kumon H, Matsui H.
    • Journal Title

      Eur.Urol. 49(1)

      Pages: 161-168

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Truncation and activation of calcineurin A by calpain I in Alzheimer disease brain.2005

    • Author(s)
      Liu F, Grundke-Iqbal I, Iqbal K, Oda Y, tomizawa K, Gong CX.
    • Journal Title

      J.Biol.Chem. 280(45)

      Pages: 37755-37762

    • Related Report
      2005 Annual Research Report
  • [Journal Article] カルシニューリン(PP2B)の恒常的活性化機構と神経細胞死制御-カルシニューリンとカルパインのクロストーク-2004

    • Author(s)
      松井秀樹, 呉 海燕, 富澤一仁
    • Journal Title

      細胞工学 Vol.23・No.5

      Pages: 533-537

    • Related Report
      2004 Annual Research Report
  • [Journal Article] タンパク質セラピー法2004

    • Author(s)
      松下正之, 富澤一仁, 松井秀樹
    • Journal Title

      実験医学 Vol.22・No.18

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Critical role of calpain-mediated cleavage of calcineurin in excitotox neurodegeneration.2004

    • Author(s)
      Wu, H.-Y., Tomizawa, K., Matsushita, M., Matsui, H.
    • Journal Title

      J.Biol.Chem. 279

      Pages: 4929-4940

    • Related Report
      2004 Annual Research Report
  • [Journal Article] HIV-1 tat inhibits long-term potentiation and attenuates spatial learning.2004

    • Author(s)
      Li, S.-T., Matsushita, M., Tomizawa, K., Matsui, H.
    • Journal Title

      Ann.Neurol 55

      Pages: 362-371

    • Related Report
      2004 Annual Research Report
  • [Journal Article] A new cell-permeable peptide allows successful allogeneic islet transplantation in mice.2004

    • Author(s)
      Noguchi, H., Matsushita, M., Tomizawa, K., Matsui, H.
    • Journal Title

      Nature Med. 10

      Pages: 305-309

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Photo-acceleration of protein release from endosome in the protein transduction system.2004

    • Author(s)
      Matsushita, M., Tomizawa, K., Matsui, H.
    • Journal Title

      FEBS Lett. 572(1-3)

      Pages: 221-226

    • Related Report
      2004 Annual Research Report

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Published: 2004-04-01   Modified: 2016-04-21  

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