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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
2009-7-2
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pubmed:abstractText |
We report here homologous recombination (HR)-mediated gene targeting of two different genes in human iPS cells (hiPSCs) and human ES cells (hESCs). HR-mediated correction of a chromosomally integrated mutant GFP reporter gene reaches efficiencies of 0.14%-0.24% in both cell types transfected by donor DNA with plasmids expressing zinc finger nucleases (ZFNs). Engineered ZFNs that induce a sequence-specific double-strand break in the GFP gene enhanced HR-mediated correction by > 1400-fold without detectable alterations in stem cell karyotypes or pluripotency. Efficient HR-mediated insertional mutagenesis was also achieved at the endogenous PIG-A locus, with a > 200-fold enhancement by ZFNs targeted to that gene. Clonal PIG-A null hESCs and iPSCs with normal karyotypes were readily obtained. The phenotypic and biological defects were rescued by PIG-A transgene expression. Our study provides the first demonstration of HR-mediated gene targeting in hiPSCs and shows the power of ZFNs for inducing specific genetic modifications in hiPSCs, as well as hESCs.
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pubmed:grant |
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pubmed:commentsCorrections |
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http://linkedlifedata.com/resource/pubmed/commentcorrection/19540188-9804556
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
1875-9777
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pubmed:author |
pubmed-author:ChenGuibinG,
pubmed-author:ChengLinzhaoL,
pubmed-author:ChouBin-KuanBK,
pubmed-author:DaleyGeorge QGQ,
pubmed-author:JoungJ KeithJK,
pubmed-author:MaederMorgan LML,
pubmed-author:MaliPrashantP,
pubmed-author:ParkIn-HyunIH,
pubmed-author:PorteusMatthew HMH,
pubmed-author:Pruett-MillerShondra MSM,
pubmed-author:Thibodeau-BegannyStaceyS,
pubmed-author:YeZhaohuiZ,
pubmed-author:ZouJizhongJ
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pubmed:issnType |
Electronic
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pubmed:day |
2
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pubmed:volume |
5
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
97-110
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pubmed:dateRevised |
2011-6-14
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pubmed:meshHeading |
pubmed-meshheading:19540188-Cell Line,
pubmed-meshheading:19540188-Deoxyribonucleases,
pubmed-meshheading:19540188-Embryonic Stem Cells,
pubmed-meshheading:19540188-Gene Targeting,
pubmed-meshheading:19540188-Genes, Reporter,
pubmed-meshheading:19540188-Green Fluorescent Proteins,
pubmed-meshheading:19540188-Humans,
pubmed-meshheading:19540188-Membrane Proteins,
pubmed-meshheading:19540188-Pluripotent Stem Cells,
pubmed-meshheading:19540188-Zinc Fingers
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pubmed:year |
2009
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