이성현 (기초과학연구원, Edgene Inc., 성균관대학교) | 한빛사논문 > 한빛사

한빛사논문

조회862

기초과학연구원, Edgene Inc., 성균관대학교

CV File 78 kb

Cell, Jan 04 2024, 187 (1) S0092-8674(23)01321-1 | https://doi.org/10.1016/j.cell.2023.11.035 Engineering TALE-linked deaminases to facilitate precision adenine base editing in mitochondrial DNA

Sung-Ik Cho ^{1 2 3 17}, Kayeong Lim ^{1 4 5 17}, Seongho Hong ^{6 7 17}, Jaesuk Lee ^{1 2}, Annie Kim ¹, Chae Jin Lim ⁸, Seungmin Ryou⁸, Ji Min Lee ^{1 2}, Young Geun Mok ^{1 9}, Eugene Chung ¹, Sanghun Kim ^{6 10}, Seunghun Han ^{6 11}, Sang-Mi Cho ⁶, Jieun Kim ^{6 7,} Eun-Kyoung Kim ⁶, Ki-Hoan Nam ⁶, Yeji Oh ⁶, Minkyung Choi ¹, Tae Hyeon An ^{12 13}, Kyoung-Jin Oh ^{12 13}, Seonghyun Lee ^{1 8 14 15 *}, Hyunji Lee ^{6 7 *}, Jin-Soo Kim ^{8 16 18 *}

¹Center for Genome Engineering, Institute for Basic Science, Daejeon 34126, Republic of Korea

²Department of Chemistry, Seoul National University, Seoul 08826, Republic of Korea

³Department of Pharmacology, Yonsei University College of Medicine, Seoul 03722, Republic of Korea

⁴Brain Science Institute, Korea Institute of Science and Technology, Seoul 02792, Republic of Korea

⁵Division of Bio-Medical Science & Technology, KIST School, Korea University of Science and Technology, Seoul 02792, Republic of Korea

⁶Laboratory Animal Resource and Research Center, Korea Research Institute of Bioscience and Biotechnology, Cheongju 28116, Republic of Korea

⁷Department of Medicine, Korea University College of Medicine, Seoul 02708, Republic of Korea

⁸Edgene, Inc., Seoul 08790, Republic of Korea

⁹GreenGene Inc., Seoul 08790, Republic of Korea

¹⁰College of Veterinary Medicine and Research Institute of Veterinary Medicine, Chungbuk National University, Cheongju 28644, Republic of Korea

¹¹Department of Biochemistry, College of Natural Sciences, Chungbuk National University, Cheongju 28644, Republic of Korea

¹²Metabolic Regulation Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea

¹³Department of Functional Genomics, KRIBB School of Bioscience, University of Science and Technology (UST), Daejeon, Republic of Korea

¹⁴Department of MetaBioHealth, Sungkyunkwan University (SKKU), Suwon, Republic of Korea

¹⁵Department of Precision Medicine, School of Medicine, Sungkyunkwan University (SKKU), Suwon, Republic of Korea

¹⁶NUS Synthetic Biology for Clinical & Technological Innovation (SynCTI) and Department of Biochemistry, National University of Singapore, Singapore, Singapore

¹⁷These authors contributed equally

¹⁸Lead contact

^*Corresponding authors: correspondence to Seonghyun Lee, Hyunji Lee or Jin-Soo Kim

Abstract

DddA-derived cytosine base editors (DdCBEs) and transcription activator-like effector (TALE)-linked deaminases (TALEDs) catalyze targeted base editing of mitochondrial DNA (mtDNA) in eukaryotic cells, a method useful for modeling of mitochondrial genetic disorders and developing novel therapeutic modalities. Here, we report that A-to-G-editing TALEDs but not C-to-T-editing DdCBEs induce tens of thousands of transcriptome-wide off-target edits in human cells. To avoid these unwanted RNA edits, we engineered the substrate-binding site in TadA8e, the deoxy-adenine deaminase in TALEDs, and created TALED variants with fine-tuned deaminase activity. Our engineered TALED variants not only reduced RNA off-target edits by >99% but also minimized off-target mtDNA mutations and bystander edits at a target site. Unlike wild-type versions, our TALED variants were not cytotoxic and did not cause developmental arrest of mouse embryos. As a result, we obtained mice with pathogenic mtDNA mutations, associated with Leigh syndrome, which showed reduced heart rates.

논문정보

형식Research article
게재일2024년 01월 (BRIC 등록일 2024-01-08)
연구진국내(교신)+국외 연구진
분야 생명과학 > 생화학

댓글 작성 로그인

CV 열람하기

연락처 보기