Generation of a tightly regulated all-cis beta cell-specific tetracycline-inducible vector.
Ability to induce protein expression at will in a cell is a powerful strategy used by scientists to better understand the function of a protein of interest. Various inducible systems have been designed in eukaryotic cells to achieve this goal. Most of them rely on two distinct vectors, one encoding a protein that can regulate transcription by binding a compound X, and one hosting the cDNA encoding the protein of interest placed downstream of promoter sequences that can bind the protein regulated by compound X (e.g., tetracycline, ecdysone). The commercially available systems are not designed to allow cell- or tissue-specific regulated expression. Additionally, although these systems can be used to generate stable clones that can be induced to express a given protein, extensive screening is often required to eliminate the clones that display poor induction or high basal levels. In the present report, we aimed to design a pancreatic beta cell-specific tetracycline-inducible system. Since the classical two-vector based tetracycline-inducible system proved to be unsatisfactory in our hands, a single vector was eventually designed that allowed tight beta cell-specific tetracycline induction in unselected cell populations.
Bulat
Natasa
N
Lausanne University, Lausanne, Switzerland.
Widmann
Christian
C
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D020690:GTPase-Activating Proteins / Q000737:chemistry / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D005821:Genetic Techniques; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D050417:Insulin-Secreting Cells / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D009928:Organ Specificity / Q000187:drug effects; D010957:Plasmids / Q000235:genetics; D020218:Response Elements / Q000235:genetics; D013752:Tetracycline / Q000494:pharmacology*
D020690:GTPase-Activating Proteins; D013752:Tetracycline