Journal article

Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases

Hockemeyer D, Soldner F, Beard C, Gao Q, Mitalipova M, DeKelver R, Katibah G, Amora R, Boydston E, Zeitler B, Meng X, Miller J, Zhang L, Rebar E, Gregory P, Urnov F, Jaenisch R ...see all

Nature Biotechnology, vol. 27, issue 9 (2009) pp. 851-857

  • 757

    Readers

    Mendeley users who have this article in their library.
  • 615

    Citations

    Citations of this article.
Sign in to save reference

Abstract

Realizing the full potential of human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) requires efficient methods for genetic modification. However, techniques to generate cell type-specific lineage reporters, as well as reliable tools to disrupt, repair or overexpress genes by gene targeting, are inefficient at best and thus are not routinely used. Here we report the highly efficient targeting of three genes in human pluripotent cells using zinc-finger nuclease (ZFN)-mediated genome editing. First, using ZFNs specific for the OCT4 (POU5F1) locus, we generated OCT4-eGFP reporter cells to monitor the pluripotent state of hESCs. Second, we inserted a transgene into the AAVS1 locus to generate a robust drug-inducible overexpression system in hESCs. Finally, we targeted the PITX3 gene, demonstrating that ZFNs can be used to generate reporter cells by targeting non-expressed genes in hESCs and hiPSCs.

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Get full text

Authors

  • Dirk Hockemeyer

  • Frank Soldner

  • Caroline Beard

  • Qing Gao

  • Maisam Mitalipova

  • Russell C DeKelver

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free