Skip to content

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

by Dirk Hockemeyer, Frank Soldner, Caroline Beard, Qing Gao, Maisam Mitalipova, Russell C DeKelver, George E Katibah, Ranier Amora, Elizabeth A Boydston, Bryan Zeitler, Xiangdong Meng, Jeffrey C Miller, Lei Zhang, Edward J Rebar, Philip D Gregory, Fyodor D Urnov, Rudolf Jaenisch show all authors
Nature biotechnology ()
Get full text at journal


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.

Cite this document (BETA)

Authors on Mendeley

Readership Statistics

722 Readers on Mendeley
by Discipline
76% Agricultural and Biological Sciences
10% Biochemistry, Genetics and Molecular Biology
9% Medicine and Dentistry
by Academic Status
34% Researcher
29% Student > Ph. D. Student
9% Student > Master
by Country
4% United States
1% United Kingdom
1% Germany

Sign up today - FREE

Mendeley saves you time finding and organizing research. Learn more

  • All your research in one place
  • Add and import papers easily
  • Access it anywhere, anytime

Start using Mendeley in seconds!

Sign up & Download

Already have an account? Sign in