Elsevier

Plasmid

Volume 69, Issue 1, January 2013, Pages 96-103
Plasmid

Construction of a doxycycline inducible adipogenic lentiviral expression system

https://doi.org/10.1016/j.plasmid.2012.10.001Get rights and content
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Abstract

To provide a tool for research on regulating adipocyte differentiation, tetracycline inducible (Tet on) lentiviral expression vectors under the control of an adipose-specific promoter were constructed. The lowest basal expression in the absence of doxycycline and most efficient dose-dependent, doxycycline-induced transient overexpression was observed using vectors constructed with a combination of Tetracycline Responsive Element (TRE) and reverse tetracycline-controlled TransActivator advanced (rtTAadv), transfected in white (3T3-L1) and brown (HIB-1B) preadipocytes cell lines. The results demonstrate that doxycycline adipogenic inducible expression can be achieved using a pLenti TRE / rtTA adv under the control of the truncated aP2 promoter in HIB-1B preadipocytes.

Highlights

► We constructed a lentiviral Tet On overexpression plasmid controlled by aP2 promoter. ► The combination of TRE + rtTAadv in vectors resulted in the lowest basal expression. ► TRE + rtTAadv also gave the best inducibility by Dox in HIB-1B and 3T3-L1 cell lines. ► A truncated aP2 promoter conferred adipogenic expression in HIB-1B cells.

Keywords

Tetracycline inducible expression
Lentiviral plasmid
Adipose-specific promoter
Adipocyte

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1

Inserm, U1016, Institut Cochin, Paris, France