The transfection of embryonic stem cells with Tet-on system and its responsiveness to doxycycline

Sci China C Life Sci. 2005 Aug;48(4):307-10. doi: 10.1360/062005-10.

Abstract

We transiently transfected pTet-on and pTRE2hyg-luciferase into the mouse embryonic stem cells (ESCs) using lipofectamine, and analyzed its inductive effect by adding serial concentrations of doxycycline (DOX). The results showed that in the transfected group, the luciferase activity of the cells was gradually increased along with the increasing concentration of DOX. While in the non-transfected group, the luciferase activity was not detectable even with DOX treatment. This indicated that the ESCs transfected with Tet-on system could response to DOX very well, and the regulation of target gene expression is dose dependent.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Carrier Proteins / genetics
  • Cells, Cultured
  • Doxycycline / pharmacology*
  • Drug Resistance, Bacterial
  • Embryonic Stem Cells / drug effects*
  • Embryonic Stem Cells / metabolism*
  • Embryonic Stem Cells / microbiology
  • Escherichia coli / genetics
  • Gene Expression Regulation / drug effects*
  • Gene Expression Regulation / physiology
  • Genes, Reporter
  • Genetic Vectors
  • Luciferases / biosynthesis
  • Luciferases / genetics
  • Mice
  • Plasmids
  • Repressor Proteins / genetics*
  • Tetracycline / pharmacology
  • Transfection* / methods

Substances

  • Bacterial Proteins
  • Carrier Proteins
  • Repressor Proteins
  • Tet O resistance protein, Bacteria
  • tetracycline resistance-encoding transposon repressor protein
  • Luciferases
  • Tetracycline
  • Doxycycline