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Open Access Highly Accessed Research article

Generation and characterization of a Tet-On (rtTA-M2) transgenic rat

Yi Sheng12, Chih-Cheng Lin2, Junming Yue123, Meena Sukhwani2, Jennifer J Shuttleworth2, Tianjiao Chu12 and Kyle E Orwig12*

Author Affiliations

1 Department of Obstetrics, Gynecology and Reproductive Sciences, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA

2 Magee-Womens Research Institute, Pittsburgh, PA 15213, USA

3 Current address: Department of Physiology, University of Tennessee, Memphis, TN 38163, USA

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BMC Developmental Biology 2010, 10:17  doi:10.1186/1471-213X-10-17

Published: 16 February 2010

Abstract

Background

The tetracycline-inducible gene regulation system is a powerful tool that allows temporal and dose-dependent regulation of target transgene expression in vitro and in vivo. Several tetracycline-inducible transgenic mouse models have been described with ubiquitous or tissue-specific expression of tetracycline-transactivator (tTA), reverse tetracycline-transactivator (rtTA) or Tet repressor (TetR). Here we describe a Tet-On transgenic rat that ubiquitously expresses rtTA-M2 driven by the murine ROSA 26 promoter.

Results

The homozygous rat line (ROSA-rtTA-M2) generated by lentiviral vector injection, has a single integration site and was derived from the offspring of a genetic mosaic founder with multiple transgene integrations. The rtTA-M2 transgene integrated into an intron of a putative gene on chromosome 2 and does not appear to affect the tissue-specificity or expression of that gene. Fibroblasts from the ROSA-rtTA-M2 rats were transduced with a TetO7/CMV-EGFP lentivirus and exhibited doxycycline dose-dependent expression of the EGFP reporter transgene, in vitro. In addition, doxycycline-inducible EGFP expression was observed, in vivo, when the TetO7/CMV-EGFP lentivirus was injected into testis, kidney and muscle tissues of ROSA-rtTA-M2 rats.

Conclusions

This conditional expression rat model may have application for transgenic overexpression or knockdown studies of gene function in development, disease and gene therapy.