Reporter Mouse Model
The reporter mouse model introduces a reporter gene such as GFP, lacZ, mCherry, tdTomato, YFP and DsRed into the locus of a target gene in order to monitor the expression of that gene at the transcriptional or translational level, and also is used to monitor protein trafficking.
The reporter mouse model is useful in monitoring the in vivo transcriptional regulation of the interested gene, they have a wide range of application prospects in cancer therapy, drug discovery, etc. For example, Cytochrome P450 3A4 (CYP3A4) plays an important role in drug metabolism, and the enzymatic activity of CYP3A4 contributes to many adverse drug-drug interactions. We make a reporter mouse model which is useful in monitoring the in vivo transcriptional regulation of the human CYP3A4 gene. A reporter construct consisting of 13 kb of the human CYP3A4 promoter controlling the firefly luciferase gene is used to generate a reporter mouse model. We detect the drug response of the reporter mouse model by injecting the different drugs resulted in a time-dependent induction of luciferase expression. The greatest and the lowest induction is found with drugs injection. It suggests that the reporter mouse model can be used to study transcriptional regulation of the gene in vivo.
That reporter mouse model has both strengths and weaknesses when compared with in vitro models. If the purpose is to predict the response of the gene to the drugs, then the results found in the reporter mouse may not be entirely predictive since the induction pattern relies upon the mouse transcription patterns. These differences in response may be due to differences in affinity for specific chemicals. There are approaches for humanizing this model which would lead to a prediction of the human response from a model.
Despite these limitations of the reporter mouse model, there are also many advantages compared with in vitro models. In vitro models, they provide no biodistribution information, and the kinetics of gene induction in vitro is not relevant to an in vivo environment. Reporter mouse model provided by Creative Animodel is highly sensitive and allows one to follow the response of the same animals over time, providing a kinetic readout in vivo.
In summary, we describe an in vivo reporter mouse model that allows us to examine the kinetics of transcriptional regulation of the gene in vivo in real time.
Creative Animodel offers a full service including vector design,construction and finishing with the generation of reporter mouse models. Besides mouse models, we also provide rat/rabbit reporter animal models. If you are interested in our laboratory, please feel free to contact us. Our research scientists will work closely with you to develop detailed study protocols to meet your goals.
The Workflow of Reporter Mouse Model
1. Reporter mouse strategy design
* Our research scientists will work closely with you to develop detailed study protocols to meet your goals
* Targeted gene analysis and determination for the founder screening by using online prediction tools and bioinformatics software
* Targeting embryos or embryonic stem(ES) cells preparation
2. Reporter mouse vector construction
* Reporter mouse vectors to construct
* Constructed vector electroporates into targeted ES cells effectively
* PCR, Southern Blot and DNA sequencing to identify recombinant ES cells
3. Pronuclear injection or ES cell targeting
1) Pronuclear injection
2) Es cell targeting
* Recombinant ES cells for blastocyst injections
* Blastocyst implanted into pseudopregnant surrogate mouse
4．Reporter mouse screening
* Chimera mouse generation and screening
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