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In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

Embryonic Stem Cells

Embryonic Stem Cells

Embryonic stem (ES) cells are undifferentiated pluripotent cells, meaning they can produce any cell type in the body. This gives them tremendous potential in science and medicine since they can generate specific cell types for use in research or to replace body cells lost due to damage or disease.

Embryonic Stem Cells

Embryonic Stem Cells

Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
ES cells are grown in a culture medium where they can divide indefinitely, creating ES cell lines. Under certain conditions, ES cells can differentiate, either spontaneously into a variety of...

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Related Experiment Video

Updated: Jun 25, 2026

Application of Mouse Parthenogenetic Haploid Embryonic Stem Cells as a Substitute of Sperm

Application of Mouse Parthenogenetic Haploid Embryonic Stem Cells as a Substitute of Sperm

Published on: November 19, 2020

Generation of double-knockout embryonic stem cells.

Eva Wielders¹, Marleen Dekker, Hein Te Riele

¹Division of Molecular Biology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)

|March 7, 2009

Summary

Disrupting both gene alleles in mouse embryonic stem cells (ES cells) allows for early study of gene function. This method enables analysis of complete gene ablation effects within the cellular context.

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Area of Science:

* Molecular Biology
* Genetics
* Developmental Biology

Background:

* Standard gene inactivation in mouse embryonic stem cells (ES cells) typically targets a single allele.
* Homozygosity for gene ablation is usually achieved by breeding heterozygous mice.
* Studying gene function at the cellular level requires disruption of both alleles.

Purpose of the Study:

* To describe methods for achieving homozygous gene disruption in ES cells.
* To enable the study of complete gene ablation consequences within the cellular context.

Main Methods:

* Consecutive targeting of both alleles using different selectable marker genes.
* Selection of cells with chromosome duplication and loss of the wild-type allele.

Main Results:

* Established methods allow for the disruption of both gene alleles in ES cells.
* This facilitates the study of cellular phenotypes resulting from complete gene ablation.

Conclusions:

* Homozygous gene disruption in ES cells is a viable strategy for studying gene function.
* This approach complements traditional in vivo methods for analyzing gene function.