Inducible Tet-On Regulatable System-Based Gene Expression In Vivo: A Tetracycline-Induced Gene Expression System for Target Gene Transcription Activation in Mouse Model

Abstract

Source: Hubner, E. K. et al.  Constitutive and Inducible Systems for Genetic In Vivo Modification of Mouse Hepatocytes Using Hydrodynamic Tail Vein Injection. J. Vis. Exp. (2018)

This video demonstrates the tetracycline, or Tet-On, gene expression system in a mouse model. In the presence of doxycycline, a tetracycline derivative, the Tet-On system consisting of reverse tetracycline-controlled transactivator protein becomes activated and expresses the gene of interest.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Induction of Tetracycline-dependent Gene or shRNA Expression

CAUTION: Doxycycline may be harmful. Please refer to the safety data sheet.

NOTE: Depending on the type and duration of the experiment, doxycycline can be supplied in drinking water.

1. For short term experiments (<10 days) administer doxycycline via drinking water using the following protocol.
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Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
D(+)-Saccharose	Carl Roth	4621.1	For sweetening of the doxycyline solution
Doxycycline hyclate	AppliChem	A2951,0025	Activation of tetracycline-dependent expression
Injekt 10 ml Syringe	Braun	4606108V
Filtropur S 0.2	Sarstedt	831.826.001	For filtration of doxycycline