한빛사 논문
한양대학교
Yeounsun Oh1,2†, Wi‑jae Lee1,3†, Junho K. Hur4,5,6†, Woo Jeung Song7, Youngjeon Lee1, Hanseop Kim1,8, Lee Wha Gwon1,9, Young‑Hyun Kim1, Young‑Ho Park10, Chan Hyoung Kim1,11, Kyung‑Seob Lim10, Bong‑Seok Song10, Jae‑Won Huh1,12, Sun‑Uk Kim10,12, Bong‑Hyun Jun3, Cheulhee Jung2* and Seung Hwan Lee1,6,9*
1National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Cheongju, Korea. 2Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, Republic of Korea. 3Department of Bioscience and Biotechnology, Konkuk University, Seoul, Korea. 4Department of Genet‑ics, College of Medicine, Hanyang University, Seoul 04763, Republic of Korea. 5Graduate School of Biomedical Sci‑ence and Engineering, Hanyang University, Seoul 04763, Republic of Korea. 6Department of Life Science, Chung-Ang University, Seoul 06974, Republic of Korea. 7Department of Medicine, Major in Medical Genetics, Graduate School, Hanyang University, Seoul 04763, Republic of Korea. 8School of Life Sciences and Biotechnology, BK21 Plus KNU Creative BioResearch Group, Kyungpook National University, Daegu, Republic of Korea. 9Department of Biomolecular Science, KRIBB School of Bioscience, Korea University of Science and Technology, Gajeong‑dong, Yuseong‑gu, Daejeon, Republic of Korea. 10Futuristic Animal Resource & Research Center (FARRC), Korea Research Institute of Bioscience and Biotech‑nology (KRIBB), Cheongju, Korea. 11Department of Biological Sciences, Chungnam National University, Daejeon, Korea. 12Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology (UST), Daejeon, Korea.
*Correspondence
†Yeounsun Oh, Wi-jae Lee and Junho K. Hur contributed equally to this work.
Abstract
Prime editing can induce a desired base substitution, insertion, or deletion in a target gene using reverse transcriptase after nick formation by CRISPR nickase. In this study, we develop a technology that can be used to insert or replace external bases in the target DNA sequence by linking reverse transcriptase to the Francisella novicida Cas9, which is a CRISPR-Cas9 ortholog. Using FnCas9(H969A) nickase, the targeting limitation of existing Streptococcus pyogenes Cas9 nickase [SpCas9(H840A)]-based prime editing is dramatically extended, and accurate prime editing is induced specifically for the target genes in human cell lines.
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