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  Functional genomics, proteomics, and regulatory DNA analysis in isogenic settings using zinc finger nuclease-driven transgenesis into a safe harbor locus in the human genome

DeKelver, R. C., Choi, V. M., Moehle, E. A., Paschon, D. E., Hockemeyer, D., Meijsing, S. H., et al. (2010). Functional genomics, proteomics, and regulatory DNA analysis in isogenic settings using zinc finger nuclease-driven transgenesis into a safe harbor locus in the human genome. Genome Research, 20(8), 1133-1142.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-7A98-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-7A99-8
Genre: Journal Article
Alternative Title : Genome Res

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 Creators:
DeKelver, R. C., Author
Choi, V. M., Author
Moehle, E. A., Author
Paschon, D. E., Author
Hockemeyer, D., Author
Meijsing, S. H.1, Author              
Sancak, Y., Author
Cui, X., Author
Steine, E. J., Author
Miller, J. C., Author
Tam, P., Author
Bartsevich, V. V., Author
Meng, X., Author
Rupniewski, I., Author
Gopalan, S. M., Author
Sun, H. C., Author
Pitz, K. J., Author
Rock, J. M., Author
Zhang, L., Author
Davis, G. D., Author
Rebar, E. J., AuthorCheeseman, I. M., AuthorYamamoto, K. R., AuthorSabatini, D. M., AuthorJaenisch, R., AuthorGregory, P. D., AuthorUrnov, F. D., Author more..
Affiliations:
1Mechanisms of Transcriptional Regulation (Sebastiaan H. Meijsing), Dept. of Computational Molecular Biology (Head: Martin Vingron), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479641              

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 Abstract: Isogenic settings are routine in model organisms, yet remain elusive for genetic experiments on human cells. We describe the use of designed zinc finger nucleases (ZFNs) for efficient transgenesis without drug selection into the PPP1R12C gene, a "safe harbor" locus known as AAVS1. ZFNs enable targeted transgenesis at a frequency of up to 15% following transient transfection of both transformed and primary human cells, including fibroblasts and hES cells. When added to this locus, transgenes such as expression cassettes for shRNAs, small-molecule-responsive cDNA expression cassettes, and reporter constructs, exhibit consistent expression and sustained function over 50 cell generations. By avoiding random integration and drug selection, this method allows bona fide isogenic settings for high-throughput functional genomics, proteomics, and regulatory DNA analysis in essentially any transformed human cell type and in primary cells.

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Language(s): eng - English
 Dates: 2010-08
 Publication Status: Published in print
 Pages: -
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 Identifiers: eDoc: 536307
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Title: Genome Research
  Alternative Title : Genome Res
Source Genre: Journal
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Pages: - Volume / Issue: 20 (8) Sequence Number: - Start / End Page: 1133 - 1142 Identifier: ISSN: 1549-5469 (Electronic)