The transfection of embryonic stem cells with Tet-on system and its responsiveness to doxycycline.
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.
Wang
Yan
Y
Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Second Medical University, Shanghai Key Laboratory of Tissue Engineering, China.
Cong
Xiaoqian
X
Liu
Deli
D
Zhang
Wenjie
W
Cui
Lei
L
Liu
Wei
W
Cao
Yilin
Y
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics; D002478:Cells, Cultured; D004318:Doxycycline / Q000494:pharmacology*; D024881:Drug Resistance, Bacterial; D053595:Embryonic Stem Cells / Q000187:drug effects* / Q000378:metabolism* / Q000382:microbiology; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000502:physiology; D017930:Genes, Reporter; D005822:Genetic Vectors; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D010957:Plasmids; D012097:Repressor Proteins / Q000235:genetics*; D013752:Tetracycline / Q000494:pharmacology; D014162:Transfection / Q000379:methods
D001426:Bacterial Proteins; D002352:Carrier Proteins; D012097:Repressor Proteins; C083139:Tet O resistance protein, Bacteria; C034811:tetracycline resistance-encoding transposon repressor protein; D008156:Luciferases; D013752:Tetracycline; D004318:Doxycycline