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Stem Cell Reports. 2017 Mar 14;8(3):491-499. doi: 10.1016/j.stemcr.2017.01.021. Epub 2017 Feb 24.

Efficient Precision Genome Editing in iPSCs via Genetic Co-targeting with Selection.

Author information

1
Department of Biochemistry, University of Utah, Salt Lake City, UT 84112, USA.
2
Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA. Electronic address: mcdec@mail.med.upenn.edu.
3
Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
4
Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
5
PharmaCell, 6229 Maastricht, the Netherlands.
6
Division of Genetics, Department of Pediatrics, Human Molecular Genetics Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
7
Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
8
Nora Eccles Harrison CVRTI, University of Utah School of Medicine, Salt Lake City, UT 84112, USA; Division of Pediatric Cardiology, University of Utah School of Medicine, Salt Lake City, UT 83113, USA.
9
Section of Quantitative Health Sciences, Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
10
Department of Biochemistry, University of Utah, Salt Lake City, UT 84112, USA; Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA.
11
Cardiovascular Center, Medical College of Wisconsin, Milwaukee, WI 53226, USA; Department of Physiology, Medical College of Wisconsin, Milwaukee, WI 53226, USA. Electronic address: ageurts@mcw.edu.

Abstract

Genome editing in induced pluripotent stem cells is currently hampered by the laborious and expensive nature of identifying homology-directed repair (HDR)-modified cells. We present an approach where isolation of cells bearing a selectable, HDR-mediated editing event at one locus enriches for HDR-mediated edits at additional loci. This strategy, called co-targeting with selection, improves the probability of isolating cells bearing HDR-mediated variants and accelerates the production of disease models.

PMID:
28238794
PMCID:
PMC5355643
DOI:
10.1016/j.stemcr.2017.01.021
[Indexed for MEDLINE]
Free PMC Article

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