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A method for genetic modification of human embryonic stem cells using electroporation

Journal Article


Abstract


  • The ability to genetically modify human embryonic stem cells (HESCs) will be critical for their widespread use as a tool for understanding fundamental aspects of human biology and pathology and for their development as a platform for pharmaceutical discovery. Here, we describe a method for the genetic modification of HESCs using electroporation, the preferred method for introduction of DNA into cells in which the desired outcome is gene targeting. This report provides methods for cell amplification, electroporation, colony selection and screening. The protocol we describe has been tested on four different HESC lines, and takes approximately 4 weeks from electroporation to PCR screening of G418-resistant clones.

Publication Date


  • 2007

Citation


  • Costa, M., Dottori, M., Sourris, K., Jamshidi, P., Hatzistavrou, T., Davis, R., . . . Stanley, E. G. (2007). A method for genetic modification of human embryonic stem cells using electroporation. Nature Protocols, 2(4), 792-796. doi:10.1038/nprot.2007.105

Scopus Eid


  • 2-s2.0-34248569914

Start Page


  • 792

End Page


  • 796

Volume


  • 2

Issue


  • 4

Abstract


  • The ability to genetically modify human embryonic stem cells (HESCs) will be critical for their widespread use as a tool for understanding fundamental aspects of human biology and pathology and for their development as a platform for pharmaceutical discovery. Here, we describe a method for the genetic modification of HESCs using electroporation, the preferred method for introduction of DNA into cells in which the desired outcome is gene targeting. This report provides methods for cell amplification, electroporation, colony selection and screening. The protocol we describe has been tested on four different HESC lines, and takes approximately 4 weeks from electroporation to PCR screening of G418-resistant clones.

Publication Date


  • 2007

Citation


  • Costa, M., Dottori, M., Sourris, K., Jamshidi, P., Hatzistavrou, T., Davis, R., . . . Stanley, E. G. (2007). A method for genetic modification of human embryonic stem cells using electroporation. Nature Protocols, 2(4), 792-796. doi:10.1038/nprot.2007.105

Scopus Eid


  • 2-s2.0-34248569914

Start Page


  • 792

End Page


  • 796

Volume


  • 2

Issue


  • 4