Use of the Sleeping Beauty Transposon System for Stable Gene Expression in Mouse Embryonic Stem Cells
- 1 Department of Biologic and Materials Sciences, University of Michigan, Ann Arbor, MI 48109, USA
- 2 Department of Cell and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA
- ↵3Corresponding author (krullc{at}umich.edu)
INTRODUCTION
Sleeping Beauty (SB) transposon-based transfection is a two-component system consisting of a transposase and a transposon containing inverted repeat/direct repeat (IR/DR) sequences that result in precise integration into a TA dinucleotide. The transposon is designed with an expression cassette of interest flanked by IR/DRs, and SB transposase mediates stable integration and reliable long-term expression of the gene of interest. It has recently been demonstrated that SB efficiently mediates gene transfer and stable gene expression in human embryonic stem (ES) cells. Here, we describe a method for transfecting and establishing stable cell lines in mouse embryonic stem (mES) cells with the SB system.
