title: Measuring and Reducing Off-Target Activities of Programmable Nucleases Including CRISPR-Cas9.
abstract: Programmable nucleases, which include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and RNA-guided engineered nucleases (RGENs) repurposed from the type II clustered, regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) system are now widely used for genome editing in higher eukaryotic cells and whole organisms, revolutionising almost every discipline in biological research, medicine, and biotechnology. All of these nucleases, however, induce off-target mutations at sites homologous in sequence with on-target sites, limiting their utility in many applications including gene or cell therapy. In this review, we compare methods for detecting nuclease off-target mutations. We also review methods for profiling genome-wide off-target effects and discuss how to reduce or avoid off-target mutations.
authors.affiliation: Center for Genome Engineering, Institute for Basic Science, Daejeon 305-811, Korea.|University of Science and Technology, Daejeon 305-350, Korea.
authors.forename: Taeyoung
authors.initials: T
authors.lastname: Koo
authors.affiliation: The Institute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Korea.
authors.forename: Jungjoon
authors.initials: J
authors.lastname: Lee
authors.affiliation: Center for Genome Engineering, Institute for Basic Science, Daejeon 305-811, Korea.|Department of Chemistry, Seoul National University, Seoul 151-742, Korea.|University of Science and Technology, Daejeon 305-350, Korea.
authors.forename: Jin-Soo
authors.initials: JS
authors.lastname: Kim
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D004720:Endonucleases / Q000235:genetics*; D005818:Genetic Engineering; D006801:Humans
chemical_list: D004720:Endonucleases
keywords: CRISPR; Cas9; TALEN; ZFN; genome editing; off-target