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Nucleic Acids Res., Apr 10 2025, 53 (7) gkaf258 | https://doi.org/10.1093/nar/gkaf258
Autophagy induction enhances homologous recombination-associated CRISPR-Cas9 gene editing
Hye Jin Nam1,2,*,†, Jun Hee Han3,†, Jihyeon Yu4,†, Chang Sik Cho5, Dongha Kim6, Young Eun Kim7, Min Ji Kim1,2, Jeong Hun Kim5,8,9,10, Dong Hyun Jo11,*, Sangsu Bae4,9,12,*
1Therapeutics & Biotechnology Division, Korea Research Institute of Chemical Technology, Daejeon 34114, Republic of Korea
2Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon 34113, Republic of Korea
3Department of Chemistry, Hanyang University, Seoul 04673, Republic of Korea
4Medical Research Center of Genomic Medicine Institute, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
5Fight Against Angiogenesis-Related Blindness (FARB) Laboratory, Biomedical Research Institute, Seoul National University, Seoul 03080, Republic of Korea
6Department of Anatomy, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea
7Center for Bioanalysis, Division of Chemical and Medical Metrology, Korea Research Institute of Standards and Science, Daejeon 34113, Republic of Korea
8Global Excellence Center for Gene & Cell Therapy (GEC-GCT), Seoul National University Hospital, Seoul 03080, Republic of Korea
9Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
10Department of Ophthalmology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
11Department of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
12Cancer Research Institute, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
*To whom correspondence should be addressed.
1Therapeutics & Biotechnology Division, Korea Research Institute of Chemical Technology, Daejeon 34114, Republic of Korea
2Department of Medicinal Chemistry and Pharmacology, University of Science and Technology, Daejeon 34113, Republic of Korea
3Department of Chemistry, Hanyang University, Seoul 04673, Republic of Korea
4Medical Research Center of Genomic Medicine Institute, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
5Fight Against Angiogenesis-Related Blindness (FARB) Laboratory, Biomedical Research Institute, Seoul National University, Seoul 03080, Republic of Korea
6Department of Anatomy, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of Korea
7Center for Bioanalysis, Division of Chemical and Medical Metrology, Korea Research Institute of Standards and Science, Daejeon 34113, Republic of Korea
8Global Excellence Center for Gene & Cell Therapy (GEC-GCT), Seoul National University Hospital, Seoul 03080, Republic of Korea
9Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
10Department of Ophthalmology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
11Department of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
12Cancer Research Institute, Seoul National University College of Medicine, Seoul 03080, Republic of Korea
*To whom correspondence should be addressed.
Correspondence may also be addressed to Hye Jin Nam.
Correspondence may also be addressed to Dong Hyun Jo.
Correspondence may also be addressed to Sangsu Bae.
†These authors contributed equally to this work.
Correspondence may also be addressed to Dong Hyun Jo.
Correspondence may also be addressed to Sangsu Bae.
†These authors contributed equally to this work.
Abstract
CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 (CRISPR-associated protein 9)-based gene editing via homologous recombination (HR) enables precise gene correction and insertion. However, its low efficiency poses a challenge due to the predominance of nonhomologous end-joining during DNA repair processes. Although numerous efforts have been made to boost HR efficiency, there remains a critical need to devise a novel method that can be universally applied across cell types and in vivo animals, which could ultimately facilitate therapeutic treatments. This study demonstrated that autophagy induction using different protocols, including nutrient deprivation or chemical treatment, significantly improved HR-associated gene editing at diverse genomic loci in mammalian cells. Notably, interacting cofactor proteins that bind to Cas9 under the autophagic condition have been identified, and autophagy induction could also enhance in vivo HR-associated gene editing in mice. These findings pave the way for effective gene correction or insertion for in vivo therapeutic treatments.
논문정보
- Research article
- 2025년 04월 (BRIC 등록일 2025-04-18)
- 국내 연구진
- 의약학 > 유전 및 유전체의학
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