Creative BioMart’s mammalian protein expression technology is able to provide conditional protein expression service. This regular plasmid vector is designed for Cre-mediated conditional gene expression in mammalian cells and animals. Expression of the gene of interest is initially silent, but can be permanently activated by co-expression of Cre recombinase, which will excise a 3x SV40 polyadenylation sequence upstream of the gene of interest. After treatment with Cre, the expression of the gene of interest is under the control of the user-selected promoter.
This regular plasmid gene expression vector system utilizes the LoxP-Stop-LoxP (LSL) cassette to achieve Cre-mediated conditional activation of gene expression in mammalian cells and animals. The LSL cassette comprises a LoxP-flanked (aka floxed) triple repeat of the SV40 polyadenylation sequence. The user-selected promoter is placed upstream of the cassette while the user’s gene of interest is placed downstream of it. In the absence of Cre recombinase, the cassette completely blocks transcription of the gene of interest. When Cre is introduced into cells carrying this vector, the cassette is excised, allowing the user-selected promoter to drive the transcription of the gene of interest.
1. This vector system can be used in tissue culture cells
2. Particular suitable for the generation of transgenic animals.
3. When using this vector system in cell culture, antibiotic or fluorescence-based markers can be added to the vector to allow selection or visualization of transfected cells, including the isolation of cells that have permanently integrated the vector in the genome.
Stable gene activation: Treatment with Cre recombinase will permanently remove the 3x SV40 polyadenylation sequence which blocks downstream transcription. This will allow transcription of the gene of interest, driven by the promoter chosen by the user.
Technical simplicity: Delivering plasmid vectors into cells by conventional transfection is technically straightforward, and far easier than virus-based vectors which require the packaging of live virus.
Very large cargo space: Our vector can accommodate ~30 kb of total DNA. The plasmid backbone only occupies about 3 kb, leaving plenty of room to accommodate the user's sequence of interest.
High-level expression: Conventional transfection of plasmids can often result in very high copy numbers in cells (up to several thousand copies per cell). This can lead to very high expression levels of the genes carried on the vector.
Suitability for in vivo applications: While this vector system can be used in tissue culture cells, it is particularly suitable for the generation of transgenic animals for Cre-mediated conditional gene expression.
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.
