Collateral cleavage of 28s rRNA by RfxCas13d causes death of mice
Collateral cleavage of 28s rRNA by RfxCas13d causes death of mice
Summary The CRISPR-Cas13 system is an RNA-guided RNA-targeting system, and has been widely used in transcriptome engineering with potentially important clinical applications. However, it is still controversial whether Cas13 exhibits collateral activity in mammalian cells. Here, we found that knocking down gene expression using RfxCas13d in the adult brain neurons caused death of mice, which was not resulted from the loss of target gene function or off-target effects. Mechanistically, we showed that RfxCas13d exhibited collateral activity in mammalian cells, which is positively correlated with the abundance of target RNA. The collateral activity of RfxCas13d could cleave 28s rRNA into two fragments, leading to translation attenuation and activation of the ZAKα-JNK/p38-immediate early gene (IEG) pathway. These results provide new mechanistic insights into the collateral activity of RfxCas13d and warn that the biosafety of CRISPR-Cas13 system needs further evaluation before applying it to clinical treatments.
Guo Qirui、Li Qi、Guo Xuefei、Liu Shengde、Chen Luoying、Kuang Ming、Li Junhong、Wang Yibing、Guo Ying、Zhang Zeming、Wang Xiao、Xia Huawei、Liu Tongtong、Liu Qinghua、Cao Lili、Li Zhiwei、Zhao Yingchi、Xu Junjie、Wei Xuemei、Li Yunfei、Luo Yujie、Wang Guodong、Wang Fengchao、You Fuping
Institute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterNational Institute of Biological Sciences, Beijing, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua UniversityInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterDepartment of Gastrointestinal Oncology, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Peking University Cancer Hospital and InstituteInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterNational Institute of Biological Sciences, Beijing, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua UniversityNational Institute of Biological Sciences, Beijing, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua UniversityInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterNational Institute of Biological Sciences, Beijing, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua UniversityBeijing Institutes of Life Science, Chinese Academy of SciencesSchool of Life Sciences, Institute of Life Sciences and Green Development, Hebei UniversityInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterNational Institute of Biological Sciences, Beijing, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua University||College of Life Sciences, Beijing Normal UniversityInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science CenterNational Institute of Biological Sciences, Beijing, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua UniversityNational Institute of Biological Sciences, Beijing, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua UniversityInstitute of Systems Biomedicine, Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Peking University Health Science Center
基础医学生物科学研究方法、生物科学研究技术分子生物学
RfxCas13dcollateral activitydeath of mice28s rRNA breakage
Guo Qirui,Li Qi,Guo Xuefei,Liu Shengde,Chen Luoying,Kuang Ming,Li Junhong,Wang Yibing,Guo Ying,Zhang Zeming,Wang Xiao,Xia Huawei,Liu Tongtong,Liu Qinghua,Cao Lili,Li Zhiwei,Zhao Yingchi,Xu Junjie,Wei Xuemei,Li Yunfei,Luo Yujie,Wang Guodong,Wang Fengchao,You Fuping.Collateral cleavage of 28s rRNA by RfxCas13d causes death of mice[EB/OL].(2025-03-28)[2025-06-29].https://www.biorxiv.org/content/10.1101/2022.01.17.476700.点此复制
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