Controlling the Gene Expression of Corynebacterium diphtheria Toxin-A Using the Tet-On System in Mouse Embryonic Stem Cells.

Mouse Embryonic Stem Cell에서 Tetracycline-Inducible System(Tet-on System)을 이용한 Corynebacterium diphtheria Toxin-A유전자의 발현 조절

  • 박재균 (포천중문의과대학교 세포유전자치료연구소) ;
  • 임수빈 (포천중문의과대학교 세포유전자치료연구) ;
  • 송지환 (포천중문의과대학교 세포유전자치료연구소)
  • Published : 2004.03.01


Embryonic stem (ES) cells are derived from the inner cell mass of the blastocyst-stage embryos that can be propagated indefinitely and, at the same time, can be differentiated into all the cell types that constitute the body. Current research using ES cells is mainly focused on the efficient generation of specific cell types by employing optimal differentiation conditions, which often requires the genetic manipulation of ES cells. As a way of developing an efficient system to regulate foreign gene expression in ES cells, we have inserted the gene encoding Corynebacterium diphtheria toxin-A (DTA) into an autonomously induced plasmid under positive doxycycline control ('Tet-on' system). In this study, we demonstrate that this system can lead to the cell death of mouse ES cells by the induction of DTA expression when exposed to the tetracycline derivative, doxycycline. MTT assay showed that this induction resulted in the apoptosis of ES cells.


Embryonic stem (ES) cells;differentiation;Tet-on system;DTA;doxycycline;Corynebacterium diphtheria toxin-A;teratoma formation


  1. Proc. Natl. Acad. Sci. v.89 Tight control of gene expression in mammalian cells by tetracyclin responsive promoters Gossen,M.;H.Bujard
  2. Anal. Biochem. v.162 Single-step method of RNA isolation by acid guanidimium thiocyanate-phenol-chlo-roform extraction Chomezynski,P.;N.Sacchi
  3. Ann. Rev. Microbiol. v.48 Mechanisms underlying expression of Tn10-encoded tetracycline resistance Hillen,W.;C.Berens
  4. Annu. Rev. Cell Dev. Biol. v.17 Embryo-derived stem cells: of mice and men Smith, A.
  5. In stem cell abiology Embryonic stem cells Smith,A.
  6. Proc. Natl. Acad. Sci. v.87 Diphtheria toxin and its ADP-ribosyltrans-ferase-defctive homologue CRM197 possess deoxyribonuclease activity Bruce,C.;R.L.Baldwin;S.L.Lessnick;B.J.Wisnieski
  7. Genes Dev. v.2 functional dissection of VP16, the trans-activatior of herper simplex virus immediate early gene expression Triezenberg,S.J.;R.C.Kingsbury;S.L.McKnight
  8. Exp. Mol. Med. v.34 Current trends of stem cell research using pluripotent stem cells Song,J.
  9. BioTech. v.89 Efficacy of tetracycline-controlled gene expression is influenced by cell type Gossen, M.;H.Bujard
  10. Infect. Immun. v.55 Expression of a mutant, full-length form of diphtheria toxin in Eschericia coli Barbieri,J.T.;R.J.Collier
  11. Biochemistry v.5 Farnesylation of CaaX-tagged diphtheria toxin A-fragment as a measure of transfer to the cytosol Falnes,P.O.;A.Wiedlocha;A.Rapak;S.Olsnes
  12. Annu. Rev. Neurosci. v.21 Inducible gene expression in the nervous system of transgenic mice Gingrich,J;J.Roder
  13. J. Natl. Cancer Inst. v.82 Comparison of in vitro anticancer-drug-screening data generated with a tetrazolium assay versus a protein assay against a diverse panel of human tumor cell lines Rubinstein,L.V.;R.H.Shoemaker;M.r.Boyd.