Generation of a tightly regulated all-cis beta cell-specific tetracycline-inducible vector.


Autoria(s): Bulat N.; Widmann C.
Data(s)

2008

Resumo

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.

Identificador

http://serval.unil.ch/?id=serval:BIB_A6B900EAA5C2

isbn:0736-6205[print], 0736-6205[linking]

pmid:18855768

doi:10.2144/000112947

isiid:000263637400005

Idioma(s)

en

Fonte

Biotechniques, vol. 45, no. 4, pp. 411, 414, 416 passim

Palavras-Chave #Animals; COS Cells; Cercopithecus aethiops; GTPase-Activating Proteins/chemistry; GTPase-Activating Proteins/metabolism; Gene Expression Regulation/drug effects; Genetic Techniques; Genetic Vectors/genetics; Humans; Insulin-Secreting Cells/drug effects; Insulin-Secreting Cells/metabolism; Mice; Organ Specificity/drug effects; Plasmids/genetics; Response Elements/genetics; Tetracycline/pharmacology
Tipo

info:eu-repo/semantics/article

article