Ecdysone Receptor-based Singular Gene Switches for Regulated Transgene Expression in Cells and Adult Rodent Tissues.
Controlled gene expression is an indispensable technique in biomedical research. Here, we report a convenient, straightforward, and reliable way to induce expression of a gene of interest with negligible background expression compared to the most widely used tetracycline (Tet)-regulated system. Exploiting a Drosophila ecdysone receptor (EcR)-based gene regulatory system, we generated nonviral and adenoviral singular vectors designated as pEUI(+) and pENTR-EUI, respectively, which contain all the required elements to guarantee regulated transgene expression (GAL4-miniVP16-EcR, termed GvEcR hereafter, and 10 tandem repeats of an upstream activation sequence promoter followed by a multiple cloning site). Through the transient and stable transfection of mammalian cell lines with reporter genes, we validated that tebufenozide, an ecdysone agonist, reversibly induced gene expression, in a dose- and time-dependent manner, with negligible background expression. In addition, we created an adenovirus derived from the pENTR-EUI vector that readily infected not only cultured cells but also rodent tissues and was sensitive to tebufenozide treatment for regulated transgene expression. These results suggest that EcR-based singular gene regulatory switches would be convenient tools for the induction of gene expression in cells and tissues in a tightly controlled fashion.
Department of Biological Sciences, College of Bioscience and Biotechnology, Chungnam National University, Daejeon, Republic of Korea.
Seoghyun
S
Lee
Department of Dermatology, College of Medicine, Chungnam National University, Daejeon, Republic of Korea.
Kyung-Cheol
KC
Sohn
Department of Dermatology, College of Medicine, Chungnam National University, Daejeon, Republic of Korea.
Dae-Kyoung
DK
Choi
Department of Pharmacology, College of Medicine, Chungnam National University, Daejeon, Republic of Korea.
Minho
M
Won
Department of Pharmacology, College of Medicine, Chungnam National University, Daejeon, Republic of Korea.
Kyeong Ah
KA
Park
Affiliated Research (and Development) Institute, Daejeon, Republic of Korea.
Sung-Kyu
SK
Ju
Department of Pharmacology, College of Medicine, Chungnam National University, Daejeon, Republic of Korea.
Kidong
K
Kang
Comparative Biomedical Research Branch, Research Institute, National Cancer Center, Goyang, Republic of Korea.
Young-Ki
YK
Bae
Department of Pharmacology, College of Medicine, Chungnam National University, Daejeon, Republic of Korea.
Gang Min
GM
Hur
Department of Biological Sciences, College of Bioscience and Biotechnology, Chungnam National University, Daejeon, Republic of Korea.
Hyunju
H
Ro
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