Development of single-vector Tet-on inducible systems with high sensitivity to doxycycline

Jiun Shuan Chao, Chun Chieh Chao, Chu Li Chang, Yi Rong Chiu, Chiun-Jye Yuan*

*Corresponding author for this work

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Single-vector Tet-on systems were developed to enable the tight regulation of transgenes in mammalian cells with a low dosage of doxycycline. Both the regulatory and the responsive units were integrated in a single vector and separated by a short DNA segment (214 bp). In the developed single-vector Tet-on systems, a high level of expression of the transgene can be induced by doxycycline at a concentration of as low as 1 ng/ml, which is 500-1,000 times lower than that usually utilized in other Tet-on systems. The single-vector Tet-on system developed here exhibited 3.5-10.8 times greater inducibility of the transgene in response to doxycycline than did a dual-vector system from a commercial source. Further studies indicate that the basal activity of Tet-on systems depends greatly on the strength of the promoter that controls the transactivator. The basal activity of Tet-on systems was high when the transactivator that was directed by the human cytomegalovirus promoter, and it was almost undetectable when the transactivator was placed under the control of a moderate strength mouse mammary tumor virus promoter. Moreover, the introduction of selectable markers allows the developed single-vector Tet-on systems to facilitate the generation of conditional transgenic cells and animals with high inducibility, low basal activity and detrimental effects of the long-term administration of doxycycline.

Original languageEnglish
Pages (from-to)240-246
Number of pages7
JournalMolecular Biotechnology
Volume51
Issue number3
DOIs
StatePublished - 1 Jul 2012

Keywords

  • CMV
  • MMTV
  • Single-vector
  • Tet-on inducible system
  • Tetracycline
  • Transgenic cell

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