Rapid-Rosa26™ Targeting technology is ingenious’ solution to the call for custom targeted transgenic models with shorter production timelines and reduced cost, without compromising performance and quality. We utilize well-validated gene targeting strategies combined with our ready-to-go enhancing vectors to ensure delivery of your expression cassette or cDNA to the Rosa26 locus in record time.
This approach offers predictable targeting, increased accuracy, and stable gene expression without perturbing endogenous genes. You benefit from streamlined model generation, lower costs, and the flexibility to achieve constitutive or conditional gene expression, enhancing research efficiency and also reproducibility.
“We found iTL’s service to be exceptionally efficient and very well managed. We generated several mouse lines with iTL, and every step for each project was performed quickly and precisely, without any delays or problems.”– Santos Franco, PhD University of Colorado Anschutz Medical Campus
Rapid-Rosa26™ Targeting technology is ingenious’ solution to the call for custom targeted transgenic models with shorter production timelines and reduced cost, without compromising performance and quality. We utilize well-validated gene targeting strategies combined with our ready-to-go enhancing vectors to deliver your expression cassette or cDNA to the Rosa26 locus in record time.Do you have another safe harbor locus in mind to use for your mouse model other than Rosa26? Contact us to discuss your goals and the design you are interested in developing for your research.
No design is too difficult for ingenious – contact us today to discuss a new transgenic line with a large insert or complex genetic elements.
The Rosa26 locus in the mouse genome defines an animal model in which a transgene is inserted in a permissive locus, also known as a “safe harbor” which allows researchers to express genes of interest. Different gene targeting technologies (embryonic stem cells, CRISPR) are used to make specific DNA insertions at the Rosa26 locus. The transgene can be constitutively or inducibly overexpressed.
Targeting transgenes to a chosen location in the genome has a number of advantages. A single copy of the DNA construct can be inserted by targeting regions of chromatin that allow the desired development and tissue-specific expression of the transgene. The Rosa26 locus is such a genomic site that is ideal to integrate a transgene of interest for stable expression.
A transgene can simply be inserted into the Rosa26 locus for ubiquitous over-expression by means of the Rosa26 promoter or CAG promoter. The CAG promoter is a strong, ubiquitously expressing promoter which can be utilized when strong over-expression is desired.
The use of a loxP flanked stop cassette will enable you to activate gene expression in your tissue of choice through mating with a tissue-specific Cre mouse line. Or, CreERT2 mice can be utilized to provide inducible transgene expression upon treatment with Tamoxifen. This approach creates a versatile line which can overexpress your construct in any tissue for which you have a Cre line.
Generally, there are three different types of Rosa26 locus knockins. In the original design, the transgenic cDNA is expressed constitutively from the Rosa26 promoter. Specifically, a splice acceptor (SA) sequence followed by cDNA of the transgene of interest is inserted into the respective restriction site within 1st intron of the Rosa26 gene. A stop cassette, composed of a neomycin resistance gene and polyadenylation (pA) sites, are inserted upstream of the transgene. A variation of the 1st strategy for conditional transgene expression is realized by using a stop cassette flanked by two lox P sites. In this case, the presence of a stop cassette can prevent the transgene from expressing unless the Cre is expressed to remove the stop cassette via homologous recombination.
If the strength of the Rosa26 promoter is not able to achieve the desired levels of transgene expression, an exogenous or enhanced promoter could be used. An example of this type of promoter is CAG and is introduced to drive high transgene expression from the Rosa26 locus.
Rosa26 locus targeting approach allows for a very robust, high frequency of correct insertion and stable expression that is widely used by the scientific community. ingenious has generated a fast, ready to use targeting vector with a variety of genetic designs that could be utilized for different applications. If tissue-specific control is essential, ingenious has designed a Rosa26 mouse line which contains a floxed stop cassette and all of the necessary Tet-On control components. This Rosa26 mouse line enables tissue-specificity and inducibility of your cDNA without the need for transactivator lines.
Rosa26 refers to a specific site on the mouse chromosome 6. It was identified in a screen that was designed to find places in the genome where the insertion of foreign DNA would not have unpredictable results.
ROSAβgeo26 (ROSA26) refers to a locus that is widely used for achieving generalized cDNA expression in a mouse model.
A safe harbor locus is a place in the genome which allows for expression of an inserted transgene without the risk of affecting surrounding endogenous genes.
You may also be interested in:
Transgenic Mouse Models
Safe Harbor Locus (Rosa26 and Polr2a)
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