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Published on: June 15, 2017
A method for the generation of conditional gene-targeted mice
Masahiro Yamamoto1, Kiyoshi Takeda
1Department of Microbiology and Immunology, Graduate School of Medicine, Osaka University, Osaka, Japan.
Abstract:
Integrins and cell adhesion molecules are required for cell adhesion and movement, as well as various biological functions such as development or immune responses. To elucidate their in vivo functions, mice lacking these molecules have been generated and various phenotypes analyzed. In the case of embryonic lethality or a requirement for spatiotemporal ablation of these genes, conditional gene-targeted mice have also been generated. Gene targeting is a powerful tool to study the function of proteins under physiological conditions and is a widely used method. Construction of targeting vectors and selection of homologous recombinant embryonic stem cells require delicate and elaborate technical procedures. In this chapter we describe the methods to generate conventional and conditional knockout mice.
Insights
This chapter details methods for creating knockout mice, essential tools for studying gene functions in vivo. These techniques enable the analysis of integrins and cell adhesion molecules in biological processes.
Area of Science:
- Molecular Biology
- Genetics
- Immunology
Background:
- Integrins and cell adhesion molecules are crucial for cell adhesion, movement, development, and immune responses.
- Understanding their in vivo functions necessitates genetic manipulation in model organisms.
- Conventional and conditional knockout mice are key tools for these investigations.
Purpose of the Study:
- To describe methodologies for generating conventional and conditional knockout mice.
- To provide insights into the technical procedures for gene targeting.
- To facilitate the study of in vivo protein functions under physiological conditions.
Main Methods:
- Gene targeting to create mice lacking specific molecules.
- Development of targeting vectors for homologous recombination.
- Selection of genetically modified embryonic stem cells.
- Generation of both conventional and conditional knockout mouse models.
Main Results:
- Established protocols for generating gene-targeted mice.
- Demonstrated the utility of knockout mice for studying gene function in vivo.
- Provided a comprehensive guide to the technical aspects of mouse model generation.
Conclusions:
- Gene targeting is a powerful and widely used method for elucidating protein functions.
- The described methods enable the creation of valuable mouse models for biological research.
- These knockout mice are instrumental in understanding development and immune responses.
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