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Updated: Sep 27, 2026
Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells
Published on: August 23, 2014
Signal transduction study using gene-targeted embryonic stem cells
Hideki Kawasome1, Takashi Hamazaki, Tetsuo Minamino
1Otsuka Pharmaceutical Co. Ltd, Tokushima, Japan.
Abstract:
Gene targeting is one of the most powerful tools to define the role of signaling molecules in animal development and disease etiology. By using this technique, nearly 1000 knockout mice have been produced over the last two decades. Generating knockout mice, however, is a time-consuming procedure. Also, an unexpected embryonic lethality sometimes prevents us from examining the function of the gene in specific tissues. Here, we describe a convenient method to directly disrupt genes at both alleles in murine embryonic stem (ES) cells. These homozygous knockout ES cells have been shown useful to determine the role of the genes in the mediation of various cellular activities such as proliferation, differentiation, apoptosis, survival, transformation, and so on. Furthermore, with the recent advance of in vitro differentiation techniques, it is now feasible to rapidly determine the role of specific molecules in particular tissues.
Insights
Gene targeting in mice is essential for understanding development and disease. This study presents a faster method using embryonic stem (ES) cells to create homozygous knockout models for gene function studies.
Area of Science:
- Molecular Biology
- Developmental Biology
- Genetics
Background:
- Gene targeting is crucial for studying gene function in development and disease.
- Existing methods for generating knockout mice are time-consuming and can be hindered by embryonic lethality.
Purpose of the Study:
- To develop a convenient and efficient method for directly disrupting both alleles of a gene in murine embryonic stem (ES) cells.
- To enable rapid assessment of gene function in specific tissues using homozygous knockout ES cells.
Main Methods:
- Direct gene disruption at both alleles in murine embryonic stem (ES) cells.
- Utilizing in vitro differentiation techniques for rapid functional analysis.
Main Results:
- Generation of homozygous knockout ES cells.
- Demonstrated utility of these cells for studying gene roles in cellular activities (proliferation, differentiation, apoptosis, etc.).
- Feasibility of rapid tissue-specific gene function determination.
Conclusions:
- The described method offers a convenient approach to generate homozygous knockout ES cells.
- This technique facilitates efficient investigation of gene function in various cellular processes and specific tissues.
