Safe Harbor Locus (Rosa26 and Polr2a)

Precise gene targeting at your choice of safe harbor locus.
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. Gene targeting at this type of locus gives you control of copy number while avoiding the undesirable effects of random insertion transgenics, namely undesirable position effects that can cause unstable phenotypes, gene silencing, or unexpected gene expression.
At ingenious, your transgene of choice can be delivered to a safe harbor locus in the mouse genome and be constitutively or inducibly overexpressed. We use both the classic Rosa26 locus, as well as Polr2a as an alternative locus for targeting. Polr2a has been shown to be a permissive locus in neuroscience² and ingenious has applied this technology to other research fields for our clients. Targeting the Polr2a locus with your transgene may be of interest if you already have Rosa26 lines and would like to mate to another line in order to study two or more targeted transgenes at once, or if you need an alternative locus to Rosa26 for your research field.
With our well-validated gene targeting strategies combined with our pre-constructed targeting vector materials, ingenious can generate precisely targeted, custom transgenic mouse models with a variety of expression possibilities at reduced timelines and cost when compared to traditional targeted knockin models.
For more information, please contact us.
References
1) Gee JM, Smith NA, Fernandez FR, Economo MN, Brunert D, Rothermel M, Morris SC, Talbot A, Palumbos S, Ichida JM, Shepherd JD, West PJ, Wachowiak M, Capecchi MR, Wilcox KS, White JA, Tvrdik P. 2014. Imaging activity in neurons and glia with a Polr2a-based and cre-dependent GCaMP5G-IRES-tdTomato reporter mouse. Neuron 83(5): 1058-72.
Notable Client Publications Using Rosa26
1) Samant SA, Pillai VB, Gupta MP. 2021. Skeletal muscle-specific over-expression of the nuclear sirtuin SIRT6 blocks cancer-associated cachexia by regulating multiple targets. JCSM Rapid Commun 4(1): 40-56.
2) McCann JJ, Vasilevskaya IA, Poudel Neupane N, Shafi AA, McNair C, Dylgjeri E, Mandigo AC, Schiewer MJ, Schrecengost RS, Gallagher P, Stanek TJ, McMahon SB, Berman-Booty LD, Ostrander WF, Knudsen KE. 2020. USP22 functions as an oncogenic driver in prostate cancer by regulating cell proliferation and DNA repair. Cancer Res 80(3): 430-443.