The adenovirus FLEX conditional Creon gene expression vector combines Creative BioMart's highly efficient adenovirus vector system with the Cre responsive FLEXconditional gene expression system, which can help you realize the introduction of the Cre responsive FLEXswitch mediated by adenovirus into a variety of mammalian cell types. FLEXCre on switch uses two pairs of loxP mutation recombination sites on both sides of the target gene to completely inhibit gene expression in the absence of Cre, and activate gene expression after Cre dependent coding sequence inversion.
FLEX Cre-on switch consists of two pairs of heteromorphic loxP mutation recombination sites, namely loxP with wild-type sequence and lox2272 with mutation sequence. Its flanking is ORF with opposite (antisense) direction to the promoter. Cre can recognize these two loxP variants, but only the same loxP site pair can be reorganized with each other, not with any other variants. LoxP and lox2272 sites are organized alternately, with each pair in reverse parallel.
In the absence of Cre recombinase, ORF is not expressed because of its antisense orientation relative to the promoter. In the presence of Cre, loxP and lox2272 sites are recombined with other loxP and lox2272 sites respectively, resulting in ORF transformation into a meaningful direction, and one of them is removed from each pair of the same recombination sites.
This allows the promoter selected by the user to drive the transcription of the target gene. Since there are two different loxP mutation sites on both sides of ORF, no further recombination will occur even if Cre exists.
| Step | Description |
| 1 | Firstly, an adenovirus vector is constructed into a plasmid in E. coli. |
| 2 | It is then transfected into packaging cells, and the vector region between the two reverse terminal repeats (ITRS) is packaged into a live virus. For the adenovirus FLEXconditional Cre on gene expression vector, the FLEX Cre-on switch is located between the two ITRS. |
| 3 | After the virus is added to the target cell, the DNA cargo is delivered to the cell, entered the nucleus and remained in the form of free DNA, but not integrated into the host genome. |
| 4 | In the process of vector construction, FLEX Cre-on switch located between the two ITRS is introduced into the target cells together with other viral genomes. |
| 5 | After Cre mediated coding sequence inversion, Cre recombinase can activate gene expression. |
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