Format

Send to:

Choose Destination
See comment in PubMed Commons below
Stem Cells. 2009 May;27(5):1032-41. doi: 10.1002/stem.38.

Cre recombination-mediated cassette exchange for building versatile transgenic human embryonic stem cells lines.

Author information

  • 1Department of Anatomy and Neurology, School of Medicine and Public Health, Waisman Center, WiCell Institute, University of Wisconsin, Madison, WI 53705, USA.

Abstract

To circumvent the silencing effect of transgene expression in human embryonic stem cells (hESCs), we employed the Cre recombination-mediated cassette exchange strategy to target the silencing-resistant site in the genome. We have identified new loci that sustain transgene expression during stem cell expansion and differentiation to cells representing the three germ layers in vitro and in vivo. The built-in double loxP cassette in the established master hESC lines was specifically replaced by a targeting vector containing the same loxP sites, using the cell-permeable Cre protein transduction method, resulting in successful generation of new hESC lines with constitutive functional gene expression, inducible transgene expression, and lineage-specific reporter gene expression. This strategy and the master cell lines allow for rapid production of transgenic hESC lines in ordinary laboratories.

PMID:
19415769
[PubMed - indexed for MEDLINE]
PMCID:
PMC2801346
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for John Wiley & Sons, Inc. Icon for PubMed Central
    Loading ...
    Write to the Help Desk