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

被引：122

作者：

Costa, Magdaline ^{[1
]}

Dottori, Mirella ^{[2
]}

Sourris, Koula ^{[1
]}

Jamshidi, Pegah ^{[2
]}

Hatzistavrou, Tanya ^{[1
]}

Davis, Richard ^{[1
]}

Azzola, Lisa ^{[1
]}

Jackson, Steven ^{[1
]}

Lim, Sue Mei ^{[1
]}

Pera, Martin ^{[2
,3
]}

Elefanty, Andrew G. ^{[1
]}

Stanley, Edouard G. ^{[1
]}

机构：

[1] Monash Univ, STRIP 1, Monash Immunol & Stem Cell Labs, Clayton, Vic 3800, Australia

[2] Australian Stem Cell Ctr, Monash Inst Med Res, Clayton, Vic, Australia

[3] Univ So Calif, Keck Sch Med, Ctr Stem Cell & Regenerat Med, Los Angeles, CA 90089 USA

来源：

NATURE PROTOCOLS | 2007年 / 2卷 / 04期

关键词：

D O I：

10.1038/nprot.2007.105

中图分类号：

Q5 [生物化学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

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.

引用

页码：792 / 796

页数：5

共 23 条

[1]

Clonally derived human embryonic stem cell lines maintain pluripotency and proliferative potential for prolonged periods of culture [J].