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Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells
Published on: August 23, 2014
Genetic manipulation of human embryonic stem cells
Silvina Epsztejn-Litman1, Rachel Eiges
1Stem Cell Research Laboratory, Medical Genetics Unit, Shaare Zedek Medical Center, Jerusalem, Israel.
Methods in Molecular Biology (Clifton, N.J.)
|November 13, 2009
Summary
Genetic manipulation of human embryonic stem (hESCs) cells is accessible using various techniques like transfection and electroporation. These methods enable gene editing for research, advancing stem cell biology and therapeutic applications.
Area of Science:
- Stem Cell Biology
- Molecular Genetics
- Developmental Biology
Background:
- Embryonic stem (ES) cells offer unique advantages for genetic manipulation due to their pluripotency and ease of clonal expansion.
- Established genetic manipulation techniques in mouse ES cells have paved the way for similar applications in human ES (hESC) cells.
Purpose of the Study:
- To detail methodologies for the genetic manipulation of human embryonic stem cells (hESCs).
- To explore the applications of genetically modified hESCs in scientific research and potential therapies.
Main Methods:
- Describes various gene delivery and manipulation techniques including transfection, electroporation, and viral infection.
- Focuses on protocols for inserting, deleting, or altering gene expression in hESCs.
- Emphasizes critical technical details for successful genetic modification and clonal expansion.
Main Results:
- Successfully applied established genetic manipulation techniques to hESCs.
- Enabled the generation of genetically modified hESC clones.
- Facilitated the study of gene function and differentiation pathways.
Conclusions:
- Genetic manipulation of hESCs is feasible and crucial for advancing stem cell research.
- These techniques are essential for understanding human development and disease.
- The described protocols provide a foundation for future therapeutic applications using engineered hESCs.
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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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