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Mol Cells. 2015 Jun;38(6):475-81. doi: 10.14348/molcells.2015.0103. Epub 2015 May 19.

Measuring and Reducing Off-Target Activities of Programmable Nucleases Including CRISPR-Cas9.

Koo T1,2, Lee J3, Kim JS1,4,2.

Author information

1
Center for Genome Engineering, Institute for Basic Science, Daejeon 305-811, Korea.
2
University of Science and Technology, Daejeon 305-350, Korea.
3
The Institute of Molecular Biology and Genetics, Seoul National University, Seoul 151-742, Korea.
4
Department of Chemistry, Seoul National University, Seoul 151-742, Korea.

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.

KEYWORDS:

CRISPR; Cas9; TALEN; ZFN; genome editing; off-target

PMID:
25985872
PMCID:
PMC4469905
DOI:
10.14348/molcells.2015.0103
[Indexed for MEDLINE]
Free PMC Article

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