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고려대학교 의과대학

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Exp. Mol. Med., Dec 01 2023, | https://doi.org/10.1038/s12276-023-01128-4 Precise base editing without unintended indels in human cells and mouse primary myoblasts

Da Eun Yoon^1,2, Na-Rae Kim¹, Soo-Ji Park^1,2, Tae Yeong Jeong^1,2, Bokkee Eun³, Yongcheol Cho⁴, Soo-Yeon Lim⁵, Hyunji Lee^2,6, Je Kyoung Seong^5,7,8 and Kyoungmi Kim¹

¹Department of Physiology, Korea University College of Medicine, Seoul 02841, Republic of Korea.

²Department of Medicine, Korea University College of Medicine, Seoul 02841, Republic of Korea.

³Core Laboratory for Convergent Translational Research, Korea University College of Medicine, Seoul 02841, Republic of Korea.

⁴Department of Brain and Cognitive Sciences, Daegu Gyeongbuk Institute of Science and Technology (DGIST), Daegu, South Korea.

⁵Korea Mouse Phenotyping Center, Seoul National University, 08826 Seoul, Republic of Korea.

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

⁷Laboratory of Developmental Biology and Genomics, BK21 Program Plus for Advanced Veterinary Science, Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, 08826 Seoul, Republic of Korea.

⁸Interdisciplinary Program for Bioinformatics, Program for Cancer Biology, BIO-MAX/N-Bio Institute, Seoul National University, 08826 Seoul, Republic of Korea

Corresponding author : Correspondence to Kyoungmi Kim.

Abstract

Base editors are powerful tools for making precise single-nucleotide changes in the genome. However, they can lead to unintended insertions and deletions at the target sites, which is a significant limitation for clinical applications. In this study, we aimed to eliminate unwanted indels at the target sites caused by various evolved base editors. Accordingly, we applied dead Cas9 instead of nickase Cas9 in the base editors to induce accurate substitutions without indels. Additionally, we tested the use of chromatin-modulating peptides in the base editors to improve nucleotide conversion efficiency. We found that using both dead Cas9 and chromatin-modulating peptides in base editing improved the nucleotide substitution efficiency without unintended indel mutations at the desired target sites in human cell lines and mouse primary myoblasts. Furthermore, the proposed scheme had fewer off-target effects than conventional base editors at the DNA level. These results indicate that the suggested approach is promising for the development of more accurate and safer base editing techniques for use in clinical applications.

논문정보

형식Research article
게재일2023년 12월 (BRIC 등록일 2023-12-04)
연구진국내 연구진
분야 생명과학 > 유전자발현

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