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Genetic Manipulation of Human Embryonic Stem Cells

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Human Embryonic Stem Cell Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 584))

Abstract

One of the great advantages of embryonic stem (ES) cells over other cell types is their accessibility to genetic manipulation. They can easily undergo genetic modifications while remaining pluripotent and can be selectively propagated, allowing the clonal expansion of genetically altered cells in culture. Since the first isolation of ES cells in mice, many effective techniques have been developed for gene delivery and manipulation of ES cells. These include transfection, electroporation, and infection protocols, as well as different approaches for inserting, deleting, or changing the expression of genes. These methods proved to be extremely useful in mouse ES cells, for monitoring and directing differentiation, discovering unknown genes and studying their function, and are now being initiated in human ES (HESC) cells. This chapter describes the different approaches and methodologies that have been applied for the genetic manipulation of HESCs and their applications. Detailed protocols for generating clones of genetically modified HESCs by transfection, electroporation, and infection will be described, with special emphasis on the important technical details that are required for this purpose.

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Authors and Affiliations

  1. Stem Cell Research Laboratory, Medical Genetics Unit, Shaare Zedek Medical Center, Jerusalem, Israel

    Silvina Epsztejn-Litman & Rachel Eiges

Authors
  1. Silvina Epsztejn-Litman
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  2. Rachel Eiges
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Editors and Affiliations

  1. Sprott Centre for Stem Cell Research, Ottawa Hosp. Research Institute (OHRI), Smyth Road 501, Ottawa, K1H 8L6, Canada

    Kursad Turksen

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC 2006

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Epsztejn-Litman, S., Eiges, R. (2009). Genetic Manipulation of Human Embryonic Stem Cells. In: Turksen, K. (eds) Human Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 584. Humana Press. https://doi.org/10.1007/978-1-60761-369-5_21

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  • DOI: https://doi.org/10.1007/978-1-60761-369-5_21

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-368-8

  • Online ISBN: 978-1-60761-369-5

  • eBook Packages: Springer Protocols

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