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Validation of oligonucleotide-mediated gene editing

Disterer, P; Simons, JP; Owen, JS; (2009) Validation of oligonucleotide-mediated gene editing. GENE THER , 16 (6) 824 - 826. 10.1038/gt.2009.32.

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Abstract

Several independent groups have reported targeted genomic editing in mammalian cells mediated by synthetic oligonucleotides. Nevertheless, the validity of data has been disputed because of experimental artefacts, inconsistent findings and low reproducibility. Here, we describe experiments designed to meet stringent criteria and completely eliminate artefactual results. In particular, by targeting cells expressing mutated enhanced green fluorescence protein (EGFP), which allow editing measurements at the protein level, and analyzing corrected clones by Southern blotting, we rigorously excluded spontaneous reversion, contamination artefacts, false-positives, or overestimation. Our findings provide unequivocal authentication that oligonucleotide-mediated gene editing is a real, not artefactual, phenomenon-a vital starting point from which to develop the technology into practical applications. Gene Therapy (2009) 16, 824-826; doi:10.1038/gt.2009.32; published online 2 April 2009

Type: Article
Title: Validation of oligonucleotide-mediated gene editing
DOI: 10.1038/gt.2009.32
Keywords: EGFP, gene repair, gene targeting, targeted sequence alteration, STRANDED-DNA OLIGONUCLEOTIDES, SEQUENCE CORRECTION, GENERATION, CONVERSION, REPAIR
UCL classification: UCL > Provost and Vice Provost Offices
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine > Inflammation
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Pop Health Sciences > Institute of Cardiovascular Science
URI: http://discovery.ucl.ac.uk/id/eprint/106163
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