Bi-directional gene switching with the tetracycline repressor and a novel tetracycline antagonist.
We have screened a panel of tetracycline (tc)-like compounds for their potential use with tc-repressor (tetR) based gene switches. The interaction between tc and tetR appears quite specific, as only tc itself and its close homologues anhydro-tc and doxycycline strongly inhibited DNA binding. However, a single tc-like compound, GR33076X, increased DNA binding of the tetR-VP16 fusion protein, both in eukaryotic cells and in bacteria. We provide evidence that this antagonist of tetracycline is potentially useful for accelerated gene switching, especially in whole animals.
Chrast-Balz
J
J
Geneva Biomedical Research Institute, Geneva, Switzerland.
Hooft van Huijsduijnen
R
R
D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004318:Doxycycline / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D017930:Genes, Reporter; D005812:Genes, Switch; D006367:HeLa Cells; D017405:Herpes Simplex Virus Protein Vmw65 / Q000235:genetics; D006801:Humans; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D013754:Tetracyclines / Q000494:pharmacology*
D004268:DNA-Binding Proteins; C101163:GR 33076X; D017405:Herpes Simplex Virus Protein Vmw65; D011994:Recombinant Proteins; D012097:Repressor Proteins; D013754:Tetracyclines; C034811:tetracycline resistance-encoding transposon repressor protein; C016229:4-epianhydrotetracycline; D008156:Luciferases; D004318:Doxycycline