Our sleeping beauty vector system is highly effective for inserting foreign DNA into the genome of host cells. This system is technically simple, utilizing plasmid transfection (rather than viral transduction) to permanently integrate your gene(s) of interest into the host genome.
This vector system is derived from the Tc1/mariner superfamily of transposons which were originally isolated from fish genomes and have been transpositionally inactive due to the accumulation of mutations. The sleeping beauty transposon was reconstructed by eliminating such inactivating mutations from sequences of Tc1/mariner transposons found in salmonids. The development of this synthetic transposon has provided a highly efficient transposon-based method for achieving transgenesis and insertional mutagenesis in vertebrates.
The sleeping beauty system contains two vectors, both engineered as E. coli plasmids. One vector, referred to as the helper plasmid, encodes the transposase. The other vector, referred to as the transposon plasmid, contains two inverted/direct repeats IR/DR(R) bracketing the region to be transposed. The gene to be delivered into host cells is cloned into this region.
When the helper and transposon plasmids are co-transfected into target cells, the transposase produced from the helper would recognize the two IR/DR(R)s on the transposon, and insert the flanked region including the two IR/DR(R)s into TA dinucleotide sites of the host genome.
Sleeping beauty is a class II transposon, meaning that it moves in a cut-and-paste manner, hopping from place to place without leaving copies behind. (In contrast, class I transposons move in a copy-and-paste manner.) Because the helper plasmid is only transiently transfected into host cells, it will get lost over time. With the loss of the helper plasmid, the integration of the transposon in the genome of host cells becomes permanent. If these cells are transfected with the helper plasmid again, the transposon could get excised from the genome of some cells, footprint free.
For further information about this vector system, please refer to the papers below.