Abstract
Targeted genetic engineering of human pluripotent cells is a prerequisite for exploiting their full potential. Such genetic manipulations can be achieved using site-specific nucleases. Here we engineered transcription activator–like effector nucleases (TALENs) for five distinct genomic loci. At all loci tested we obtained human embryonic stem cell (ESC) and induced pluripotent stem cell (iPSC) clones carrying transgenic cassettes solely at the TALEN-specified location. Our data suggest that TALENs employing the specific architectures described here mediate site-specific genome modification in human pluripotent cells with similar efficiency and precision as do zinc-finger nucleases (ZFNs).
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Zou, J. et al. Cell Stem Cell 5, 97–110 (2009).
Hockemeyer, D. et al. Nat. Biotechnol. 27, 851–857 (2009).
Hockemeyer, D. & Jaenisch, R. Cold Spring Harb. Symp. Quant. Biol. 75, 201–209 (2010).
Urnov, F.D., Rebar, E.J., Holmes, M.C., Zhang, H.S. & Gregory, P.D. Nat. Rev. Genet. 11, 636–646 (2010).
Soldner, F. et al. Cell (in the press).
Boch, J. & Bonas, U. Annu. Rev. Phytopathol. 48, 419–436 (2010).
Boch, J. et al. Science 326, 1509–1512 (2009).
Moscou, M.J. & Bogdanove, A.J. Science 326, 1501 (2009).
Miller, J.C. et al. Nat. Biotechnol. 29, 143–148 (2011).
Cermak, T. et al. published online, doi:10.1093/nar/gkr218 (14 April 2011).
Lengner, C.J. et al. Cell 141, 872–883 (2010).
Hockemeyer, D. et al. Cell Stem Cell 3, 346–353 (2008).
Zwaka, T. & Thomson, J. Nat. Biotechnol. 21, 319–321 (2003).
Doyon, Y. et al. Nat. Methods 8, 74–79 (2011).
Zhang, F. et al. Nat. Biotechnol. 29, 149–153 (2011).
Acknowledgements
We thank R. Alagappan, P. Xu, S. Cristea, S. Lam and A. Vincent for expert technical assistance. We thank F. Soldner for helpful discussions on the manuscript. D.H. is a Merck Fellow of the Life Sciences Research Foundation. R.J. was supported by US National Institutes of Health grants R37-CA084198, RO1-CA087869 and RO1-HD045022 and by a grant from the Howard Hughes Medical Institute.
Author information
Authors and Affiliations
Contributions
D.H., H.W. and R.J. designed the targeting experiments and wrote the manuscript. H.W. and D.H. generated donor plasmids. D.H. preformed targeting experiments. S.K., C.S.L. and H.W. assisted with Southern blot analysis. Q.G. analyzed teratomas. J.P.C. and D.H. performed FACS analysis of targeted cells. L.Z. and J.C.M. designed the TALENs, S.J.H. assembled the TALENs, G.J.C. and Y.S. tested the TALENs, and B.Z., J.M.C. and X.M. performed the off-target analysis. D.H., R.J., L.Z., G.J.C., J.C.M., B.Z., X.M. and F.D.U. analyzed the data. E.J.R., P.D.G. and F.D.U. designed and supervised the design of the TALENs and contributed to writing the manuscript.
Corresponding author
Ethics declarations
Competing interests
R.J. is an adviser to Stemgent and a cofounder of Fate Therapeutics. G.J.C., L.Z., Y.S., J.C.M., B.Z., J.M.C., X.M., S.J.H., E.J.R., P.D.G. and F.D.U. are full-time employees of Sangamo BioSciences.
Supplementary information
Supplementary Text and Figures
Supplementary Tables 1–4, Supplementary Notes and Supplementary Figures 1–10 (PDF 8120 kb)
Rights and permissions
About this article
Cite this article
Hockemeyer, D., Wang, H., Kiani, S. et al. Genetic engineering of human pluripotent cells using TALE nucleases. Nat Biotechnol 29, 731–734 (2011). https://doi.org/10.1038/nbt.1927
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nbt.1927
