Inducible point mutation is to introduce a point mutation in a time-controlled manner to generate inducible point mutant animal models, such as inducible point mutant mouse models or rat models. Scientists may also deliberately introduce mutant sequences through DNA manipulation for the sake of scientific experimentation. This strategy allows them to bypass the limits such as lethal phenotype, compensatory mechanisms and complex phenotype, usually observed with constitutive point mutation.
Inducible Point Mutant Mouse Model
There have been some inducible point mutant mouse models reported.For example, for the study of acute myeloid leukemia(AML) which is a major hematopoietic malignancy characterized by the proliferation of a malignant clone of myeloid progenitor cells, AML1-ETO knockin mice were generated to study the role of AML1-ETO in leukemogenesis. This knockin mice died in midgestation because of hemorrhaging in the central nervous system and a block of definitive hematopoiesis during embryogenesis. So they were not good animal models for the study of AML. Therefore, mouse models were generated when the expression of AML1-ETO is under the control of a tetracycline-inducible system. Inducible point mutant mice have been generated with the AML1-ETO complementary DNA controlled by a tet-responsive element. In the absence of the tetracycline, AML1-ETO is strongly expressed in the bone marrow of AML1-ETO and tetracycline-controlled transcriptional activator double-positive inducible point mutant mice. Furthermore, the addition of tetracycline reduces AML1-ETO expression in double-positive inducible knockin mice to nondetectable levels.
Creative Animodel has constructed systematic data and experience in generating inducible point mutant mouse and rat 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 Inducible Point Mutant Mouse Model
1. Inducible Point Mutant 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
* Targeted embryonic stem(ES) cells preparation
2. Inducible Point Mutant Mouse Vector Construct
* Point mutant 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
* Recombinant ES cells for blastocyst injections
* Blastocyst implanted into pseudopregnant surrogate mouse
4．Inducible Point Mutant Mouse Screening
* Inducible point mutant mouse generation and screening
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