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Abstract
The CRISPR-Cas nickase system for genome editing has attracted considerable attention owing to its safety, efficiency, and versatility. Although alternative effectors to Cas9 have the potential to expand the scope of genome editing, their application has not been optimized. Herein, we used an enhanced CRISPR-Cas12a nickase system to induce mutations by targeting genes in a human-derived cell line. The optimized CRISPR-Cas12a nickase system effectively introduced mutations into target genes under a specific directionality and distance between nickases. In particular, the single-mode Cas12a nickase system can induce the target-specific mutations with less DNA double-strand breaks. By inducing mutations in the Thymine-rich target genes in single- or dual-mode, Cas12a nickase compensates the limitations of Cas9 nickase and is expected to contribute to the development of future genome editing technologies.
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1 Korea Research Institute of Bioscience and Biotechnology (KRIBB), National Primate Research Center (NPRC), Cheongju, Republic of Korea (GRID:grid.249967.7) (ISNI:0000 0004 0636 3099); Chungnam National University, Department of Biological Sciences, Daejeon, Republic of Korea (GRID:grid.254230.2) (ISNI:0000 0001 0722 6377)
2 Chung-Ang University, Department of Life Science, Seoul, Republic of Korea (GRID:grid.254224.7) (ISNI:0000 0001 0789 9563)
3 Korea Research Institute of Bioscience and Biotechnology (KRIBB), National Primate Research Center (NPRC), Cheongju, Republic of Korea (GRID:grid.249967.7) (ISNI:0000 0004 0636 3099); Korea University of Science and Technology (UST), Department of Functional Genomics, KRIBB School of Bioscience, Daejeon, Republic of Korea (GRID:grid.412786.e) (ISNI:0000 0004 1791 8264)
4 Hanyang University, Department of Medicine, Major in Medical Genetics, Graduate School, Seoul, Republic of Korea (GRID:grid.49606.3d) (ISNI:0000 0001 1364 9317)
5 Korea Research Institute of Bioscience and Biotechnology (KRIBB), National Primate Research Center (NPRC), Cheongju, Republic of Korea (GRID:grid.249967.7) (ISNI:0000 0004 0636 3099)
6 Korea Research Institute of Bioscience and Biotechnology (KRIBB), Futuristic Animal Resource and Research Center (FARRC), Cheongju, Republic of Korea (GRID:grid.249967.7) (ISNI:0000 0004 0636 3099)
7 Chungnam National University, Department of Biological Sciences, Daejeon, Republic of Korea (GRID:grid.254230.2) (ISNI:0000 0001 0722 6377)
8 Hanyang University, Department of Genetics, College of Medicine, Seoul, Republic of Korea (GRID:grid.49606.3d) (ISNI:0000 0001 1364 9317); Hanyang University, Graduate School of Biomedical Science and Engineering, Seoul, Republic of Korea (GRID:grid.49606.3d) (ISNI:0000 0001 1364 9317)