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Themes:
Themes:
1. Genomic imprinting (Igf2/H19 genes)
1. Genomic imprinting (Igf2/H19 genes)
2. Hypertension-responsive gene regulation (renin)
2. Hypertension-responsive gene regulation (renin)
Research:
Research:
Many eukaryotic genes (such as globin, hox, imprinted genes, etc.) are organized into gene clusters and the coordinated expression of these genes during development is critical for their functions. The temporal and spatial control of gene expression at such loci is known to involve long-range chromatin interactions through cis-regulatory elements (such as enhancers, imprinting control regions (ICRs) and insulator sites) and the establishment of preferred epigenetic states (DNA and histone modifications). However, how these factors co-ordinate complex gene expression patterns is not well understood. The focus of our laboratory is to analyze the mechanistic basis of such regulation and we have studied the beta-globin, renin (both to elucidate the enhancer-promoter interaction mechanism) and Igf2/H19 gene loci (for the mono-allelic gene expression mechanism). To analyze long-range gene expression mechanisms, we have introduced non-conventional vector systems capable of cloning a large DNA insert (i.e. YAC and BAC) and a recently developed CRISPR/Cas genome editing system to generate transgenic and knock-out/in model organisms, respectively. As genomic imprinting is a transgenerational phenomenon unique to mammals, and renin gene expression in the kidney is regulated by blood pressure and body fluid volume, it is also important to use a host system that allows both temporal and spatial assessment of gene expression, and a mouse genetic system is appropriate for this purpose. We use biochemical, molecular biology, and cell culture techniques in combination with mouse molecular genetics to identify and functionally characterize cis-regulatory DNA elements and trans-acting components involved in the regulation. (A partial list is shown below)
Many eukaryotic genes (such as globin, hox, imprinted genes, etc.) are organized into gene clusters and the coordinated expression of these genes during development is critical for their functions. The temporal and spatial control of gene expression at such loci is known to involve long-range chromatin interactions through cis-regulatory elements (such as enhancers, imprinting control regions (ICRs) and insulator sites) and the establishment of preferred epigenetic states (DNA and histone modifications). However, how these factors co-ordinate complex gene expression patterns is not well understood. The focus of our laboratory is to analyze the mechanistic basis of such regulation and we have studied the beta-globin, renin (both to elucidate the enhancer-promoter interaction mechanism) and Igf2/H19 gene loci (for the mono-allelic gene expression mechanism). To analyze long-range gene expression mechanisms, we have introduced non-conventional vector systems capable of cloning a large DNA insert (i.e. YAC and BAC) and a recently developed CRISPR/Cas genome editing system to generate transgenic and knock-out/in model organisms, respectively. As genomic imprinting is a transgenerational phenomenon unique to mammals, and renin gene expression in the kidney is regulated by blood pressure and body fluid volume, it is also important to use a host system that allows both temporal and spatial assessment of gene expression, and a mouse genetic system is appropriate for this purpose. We use biochemical, molecular biology, and cell culture techniques in combination with mouse molecular genetics to identify and functionally characterize cis-regulatory DNA elements and trans-acting components involved in the regulation. (A partial list is shown below)
Host organisms:
Host organisms:
- microorganisms (E. coli, Yeast)
- microorganisms (E. coli, Yeast)
- Mammalian cultured cells (Cancer cells, ES cells, MEF cells, etc.）
- Mammalian cultured cells (Cancer cells, ES cells, MEF cells, etc.）
- Mouse
- Mouse
Techniques (biochemistry, molecular biology and cell culture):
Techniques (biochemistry, molecular biology and cell culture):
- Generation of transgenic mice by pronuclear DNA injection
- Generation of transgenic mice by pronuclear DNA injection
- Gene targeting in mouse embryonic stem (ES) cell
- Gene targeting in mouse embryonic stem (ES) cell
- Generation of chimeric mice (by blastocyst injection or by aggregating of ES cells with 8-cell stage embryos)
- Generation of chimeric mice (by blastocyst injection or by aggregating of ES cells with 8-cell stage embryos)
- Generation of gene knock-out/ knock-in mice
- Generation of gene knock-out/ knock-in mice
- Genome editing by CRISPR/Cas9 system (cultured cells and mice)
- Genome editing by CRISPR/Cas9 system (cultured cells and mice)
- In vitro fertilization of mouse embryos
- In vitro fertilization of mouse embryos
- Freezing mouse embryos for long term stock in LN2
- Freezing mouse embryos for long term stock in LN2
Recruitment:
Recruitment:
Graduate School of Science and Technology, Degree Programs in Life and Earth Sciences, University of Tsukuba
Graduate School of Science and Technology, Degree Programs in Life and Earth Sciences, University of Tsukuba
Degree Program in Agro-Bioresources Science and Technology (Master's program)
Degree Program in Agro-Bioresources Science and Technology (Master's program)
Degree Program in Life and Agricultural Sciences (Doctor's program)
Degree Program in Life and Agricultural Sciences (Doctor's program)
Contact information:
Contact information:
Keiji Tanimoto, Ph.D.
Keiji Tanimoto, Ph.D.
Faculty of Life and Environmental Sciences
Faculty of Life and Environmental Sciences
Life Science Center, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, Tsukuba, Ibaraki, 305-8577, Japan
Life Science Center, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, Tsukuba, Ibaraki, 305-8577, Japan
e-mail: keiji[at]tara.tsukuba.ac.jp ([at] should be replaced with @)
e-mail: keiji[at]tara.tsukuba.ac.jp ([at] should be replaced with @)
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