Genetically-modified laboratory animals are of great importance in biomedical research. The Sleeping Beauty (SB) transposon system, an insertional mutagenesis system, represents an important tool for genetic studies. The aim of this study protocol is to describe a method for inducible expression of the transposase upon administration of doxycycline using the Tet-on system. This protocol requires three strains of transgenic mice. The first mouse expresses the rtTA (reverse tetracycline trans-activator gene) using a tissue specific promoter (TS-rtTA mouse). The second transgenic mouse expresses the SB transposase cDNA and GFP under control of the TRE promoter (TRE-SB-IRES-GFP mouse). The third mouse contains the T2/Onc3 concatamer of transposons (T2/Onc3 Mouse). The three strains are interbred to get mice that contain all three elements (TS-rtTA, TRE-SB-IRES-GFP, and T2/Onc3). Administration of doxycycline will activate transcription of TRE-SB-IRES-GFP only in cells that are expressing rtTA, which will activate T2/Onc3.
|Original language||English (US)|
|Journal||Annals of Tropical Medicine and Public Health|
|Issue number||Special Issue|
|State||Published - Jan 1 2018|