According to the certainty of gene site, it can be divided into:
Targeted conditional overexpression
Rosa26 is one of the most frequently used sites for site-specific integration. It is a non-coding gene located on mouse chromosome 6 that has been shown to be expressed in most tissues and cell types. As the widespread expression of gene requires the binding of transcription factors, the genomic structure will not be heterochromatinized and hence, exogenous genes which are inserted at this site are highly likely to be expressed in each tissue. Therefore, Rosa26 is knowns as a “safe harbor”= for the expression of exogenous genes.
Cre-LoxP system can be used to inducibly regulate exogenous gene.Generate the Rosa26-(SA/pCAG)-loxp-Stop-loxp-cDNA-pA recombinant vector and insert this conditional overexpression structure into the Rosa26 gene intron1. Hybridize such knock-in mice with a variety of Cre recombinase-expressing mice to obtain conditional overexpression mouse models with tissue-specific expression of the exogenous gene.
Frees you from multiple copies, multi-site insertion, unstable expression, the need to establish animal strains, and other problems that you may encounter with transgenic models created via random integration
Achieves stable and controllable expression of exogenous genes to help avoid unpredictable and abnormal phenotypes that may result from systemic gene overexpression
Allows for site-specific integration of super large fragments (20-30kb)
In addition to the Rosa26 site, several other sites are available to meet different research purposes
Can be used for overexpression experiments on gene and protein functions
Can be used for rescue experiments for KO phenotypes
Can be used to create expression models of humanized genes when no human homologous genes are available
Can be used to study gene mutations of human diseases
Transgenic mouse models can be obtained more efficiently using the piggyBac transposon system. Clone the target fragment into the piggyBac transposon plasmid. Inject the plasmid into the fertilized egg together with the piggyBac transposase. Transposase will integrate the target fragment into the 5'-TTAA-3' site on the genome to obtain transgenic mice.
Since the piggyBac transposon can specifically recognize the 5’-TTAA-3’ site in the genome with the help of a transposase, it can precisely cleave and insert the gene without leaving any mark. In addition, in the process of random insertion into the genome, the gene is more likely to be inserted into a position undergoing active transcription, thus greatly increasing the positive expression rate of the exogenous gene in the transgenic mice.
A quick and efficient way to obtain the Founder mice of the transgenic model
Rapid detection of target gene expression in Founder mice using a luciferase reporter gene in conjunction with an in vivo imaging system
you may contact our technical consultants to design and customize your gene overexpression mouse model.