Creative BioMart's sleeping beauty vector system is a highly effective system 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.
1.) The transposon consists of a mirrored set of inverted repeats (green) flanking the gene of interest (blue) in a plasmid backbone. A separate plasmid contains the transposase gene (red) for expressing the transposase enzyme.
2.) The transposase enzyme is expressed (red stars) and binds the inverted repeats (green), and an endonuclease reaction occurs which cuts the DNA.
3.) The released transposon can now bind a strand of DNA with a TA dinucleotide (there are many such sites in the human genome). The original plasmid is empty following the removal of the transposon; the plasmid is then degraded by the cell.
4.) The transposase creates a double-strand break in the DNA and allows the transposon to integrate. Additionally, the TA sequence is duplicated near the gene of interest insertion site. The sleeping beauty system contains two vectors, both engineered as E. coli plasmid.
A. One vector, referred to as the helper plasmid, encodes the transposase.
B. 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.
Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP. If you have any special needs or are interested in learning more about Creative BioMart's protein expression services, please feel free to inquire about more details.
