title: Quantifying on- and off-target genome editing.
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.
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA.
authors.forename: Ayal
authors.initials: A
authors.lastname: Hendel
authors.affiliation: Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
authors.forename: Eli J
authors.initials: EJ
authors.lastname: Fine
authors.affiliation: Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
authors.forename: Gang
authors.initials: G
authors.lastname: Bao
authors.affiliation: Department of Pediatrics, Stanford University, Stanford, CA 94305, USA. Electronic address: mporteus@stanford.edu.
authors.forename: Matthew H
authors.initials: MH
authors.lastname: Porteus
mesh_terms: D059766:DNA End-Joining Repair; D004720:Endonucleases / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D005818:Genetic Engineering / Q000379:methods*; D056426:Genetic Loci; D015316:Genetic Therapy / Q000379:methods*; D015894:Genome, Human; D049452:Green Fluorescent Proteins / Q000235:genetics; D059765:Homologous Recombination; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D016335:Zinc Fingers / Q000235:genetics; D000097573:Red Fluorescent Protein
chemical_list: D004720:Endonucleases; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein
keywords: CRISPR/Cas9; RNA-guided endonucleases; TALENs; ZFNs; gene editing; gene targeting; homologous recombination; nonhomologous end-joining