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Highly B lymphocyte-specific tamoxifen inducible transgene expression of CreER T2 by using the LC-1 locus BAC vector
Peter Boross1, Cor Breukel, Pieter Fokko van Loo
1Department of Human and Clinical Genetics, Leiden University Medical Center, Leiden, The Netherlands.
Abstract:
The generation of cell type specific inducible Cre transgenic mice is the most challenging and limiting part in the development of spatio-temporally controlled knockout mouse models. Here we report the generation and characterization of a B lymphocyte-specific tamoxifen-inducible Cre transgenic mouse strain, LC-1-hCD19-CreER(T2). We utilized the human CD19 promoter for expression of the tamoxifen-inducible Cre recombinase (CreER(T2)) gene, embedded in genomic sequences previously reported to give minimal position effects after transgenesis. Cre recombinase activity was evaluated by cross-breeding the LC-1-hCD19-CreER(T2) strain with a strain containing a floxed gene widely expressed in the hematopoietic system. Cre activity was only detected in the presence of tamoxifen and was restricted to B lymphocytes. The efficacy of recombination ranged from 27 to 61% in the hemizygous and homozygous mice, respectively. In conclusion, the LC-1-hCD19-CreER(T2) strain is a powerful tool to study gene function specifically in B lymphocytes at any chosen time point in the lifecycle of the mouse.
Insights
Researchers developed a new B lymphocyte-specific inducible Cre transgenic mouse model, LC-1-hCD19-CreER(T2). This tool enables precise temporal control for studying gene function in B lymphocytes.
Area of Science:
- Immunology
- Genetics
- Molecular Biology
Background:
- Generating cell type-specific inducible Cre transgenic mice is crucial for spatio-temporally controlled knockout mouse models.
- Existing methods face challenges in specificity and control, limiting research in B lymphocyte function.
Purpose of the Study:
- To develop and characterize a novel tamoxifen-inducible Cre transgenic mouse strain for B lymphocyte-specific gene manipulation.
- To enable precise temporal control over gene knockout in B lymphocytes for functional studies.
Main Methods:
- Utilized the human CD19 promoter to drive expression of Cre recombinase (CreER(T2)) in a tamoxifen-inducible manner.
- Generated the LC-1-hCD19-CreER(T2) transgenic mouse strain, embedding the gene in sequences minimizing position effects.
- Evaluated Cre recombinase activity by cross-breeding with a floxed gene reporter strain and administering tamoxifen.
Main Results:
- Cre recombinase activity was confirmed to be tamoxifen-dependent.
- Expression and activity of Cre were strictly restricted to B lymphocytes.
- Recombination efficacy ranged from 27% in hemizygous to 61% in homozygous mice.
Conclusions:
- The LC-1-hCD19-CreER(T2) mouse strain provides a powerful and specific tool for B lymphocyte research.
- This model allows for inducible and temporally controlled gene function studies in B lymphocytes.
- Facilitates advanced investigations into B cell biology and immune responses.
