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Trends Biotechnol. 2015 Feb;33(2):132-40. doi: 10.1016/j.tibtech.2014.12.001. Epub 2015 Jan 13.

Quantifying on- and off-target genome editing.

Author information

1
Department of Pediatrics, Stanford University, Stanford, CA 94305, USA.
2
Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
3
Department of Pediatrics, Stanford University, Stanford, CA 94305, USA. Electronic address: mporteus@stanford.edu.

Abstract

Genome editing with engineered nucleases is a rapidly growing field thanks to transformative technologies that allow researchers to precisely alter genomes for numerous applications including basic research, biotechnology, and human gene therapy. While the ability to make precise and controlled changes at specified sites throughout the genome has grown tremendously in recent years, we still lack a comprehensive and standardized battery of assays for measuring the different genome editing outcomes created at endogenous genomic loci. Here we review the existing assays for quantifying on- and off-target genome editing and describe their utility in advancing the technology. We also highlight unmet assay needs for quantifying on- and off-target genome editing outcomes and discuss their importance for the genome editing field.

KEYWORDS:

CRISPR/Cas9; RNA-guided endonucleases; TALENs; ZFNs; gene editing; gene targeting; homologous recombination; nonhomologous end-joining

PMID:
25595557
PMCID:
PMC4308725
DOI:
10.1016/j.tibtech.2014.12.001
[Indexed for MEDLINE]
Free PMC Article

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