한빛사 논문
Seung-Hun Kang1,2, Wi-jae Lee2,3, Ju-Hyun An2,4, Jong-Hee Lee5, Young-Hyun Kim4,5, Hanseop Kim2,6, Yeounsun Oh2,7, Young-Ho Park2, Yeung Bae Jin5, Bong-Hyun Jun3, Junho K. Hur8,9,10,*, Sun-Uk Kim2,4,* & Seung Hwan Lee5,*
1Department of Medicine, Graduate School, Kyung Hee University, Seoul, Republic of Korea.
2Futuristic Animal Resource & Research Center (FARRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Cheongju, Republic of Korea.
3Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Korea.
4Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Korea.
5National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Cheongju, Korea.
6School of Life Sciences and Biotechnology, BK21 Plus KNU Creative BioResearch Group, Kyungpook National University, Daegu, Republic of Korea.
7Division of Biotechnology, College of Life Science and Biotechnology, Korea University, Seoul 02841, Republic of Korea.
8Department of Pathology, College of Medicine, Kyung Hee University, Seoul, Republic of Korea.
9Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul, Republic of Korea.
10Department of Medical Genetics, College of Medicine, Hanyang University, Seoul, Republic of Korea.
*Corresponding author
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 (1.6~984 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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TOP52020년 후보
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