Transcriptional activation by tetracyclines in mammalian cells.
A transcriptional transactivator was developed that fuses the VP16 activation domain with a mutant Tet repressor from Escherichia coli. This transactivator requires certain tetracycline (Tc) derivatives for specific DNA binding. Thus, addition of doxycycline to HeLa cells that constitutively synthesized the transactivator and that contained an appropriate, stably integrated reporter unit rapidly induced gene expression more than a thousandfold. The specificity of the Tet repressor-operator-effector interaction and the pharmacological characteristics of Tc's make this regulatory system well suited for the control of gene activities in vivo, such as in transgenic animals and possibly in gene therapy.
Gossen
M
M
Zentrum für Molekulare Biologie der Universität Heidelberg, (ZMDH), Im Neuenheimer Feld 282, Germany.
Freundlieb
S
S
Bender
G
G
Müller
G
G
Hillen
W
W
Bujard
H
H
D000595:Amino Acid Sequence; D004318:Doxycycline / Q000494:pharmacology*; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D009875:Operator Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics*; D018139:Simplexvirus; D015534:Trans-Activators / Q000235:genetics*; D015533:Transcriptional Activation / Q000187:drug effects*; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics
D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D014764:Viral Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D004318:Doxycycline