Prediction-based highly sensitive CRISPR off-target validation using target-specific DNA enrichment
- Authors
- Kang, Seung-Hun; Lee, Wi-jae; An, Ju-Hyun; Lee, Jong-Hee; Kim, Young-Hyun; Kim, Hanseop; Oh, Yeounsun; Park, Young-Ho; Jin, Yeung Bae; Jun, Bong-Hyun; Hur, Junho K.; Kim, Sun-Uk; Lee, Seung Hwan
- Issue Date
- Jul-2020
- Publisher
- NATURE PUBLISHING GROUP
- Citation
- NATURE COMMUNICATIONS, v.11, no.1
- Journal Title
- NATURE COMMUNICATIONS
- Volume
- 11
- Number
- 1
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/63348
- DOI
- 10.1038/s41467-020-17418-8
- ISSN
- 2041-1723
- Abstract
- CRISPR effectors, which comprise a CRISPR-Cas protein and a guide (g)RNA derived from the bacterial immune system, are widely used for target-specific genome editing. When the gRNA recognizes genomic loci with sequences that are similar to the target, deleterious mutations can occur. Off-target mutations with a frequency below 0.5% remain mostly undetected by current genome-wide off-target detection techniques. Here we report a method to effectively detect extremely small amounts of mutated DNA based on predicted off-target-specific amplification. In this study, we used various genome editors to induce intracellular genome mutations, and the CRISPR amplification method detected off-target mutations at a significantly higher rate (1.6 similar to 984 fold increase) than an existing targeted amplicon sequencing method. In the near future, CRISPR amplification in combination with genome-wide off-target detection methods will allow detection of genome editor-induced off-target mutations with high sensitivity and in a non-biased manner.
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- Appears in
Collections - College of Natural Sciences > Department of Life Science > 1. Journal Articles
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