2. Academic Validation
  3. The collateral activity of RfxCas13d can induce lethality in a RfxCas13d knock-in mouse model

The collateral activity of RfxCas13d can induce lethality in a RfxCas13d knock-in mouse model

  • Genome Biol. 2023 Feb 1;24(1):20. doi: 10.1186/s13059-023-02860-w.
Yunfei Li # 1,  Junjie Xu # 2 3,  Xuefei Guo # 4,  Zhiwei Li # 5,  Lili Cao 6,  Shengde Liu 7,  Ying Guo 2,  Guodong Wang 2,  Yujie Luo 4,  Zeming Zhang 4,  Xuemei Wei 4,  Yingchi Zhao 4,  Tongtong Liu 4,  Xiao Wang 4,  Huawei Xia 4,  Ming Kuang 4,  Qirui Guo 4,  Junhong Li 4,  Luoying Chen 4,  Yibing Wang 2,  Qi Li 2,  Fengchao Wang 2,  Qinghua Liu 8,  Fuping You 9
Affiliations

Affiliations

  • 1 Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Institute of Systems Biomedicine, Peking University Health Science Center, Beijing, 100191, China. [email protected].
  • 2 National Institute of Biological Sciences, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua University, Beijing, 102206, China.
  • 3 College of Life Sciences, Beijing Normal University, Beijing, 100875, China.
  • 4 Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Institute of Systems Biomedicine, Peking University Health Science Center, Beijing, 100191, China.
  • 5 School of Life Sciences, Institute of Life Sciences and Green Development, Hebei University, Baoding, 071002, Hebei, China.
  • 6 Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing, 100101, China.
  • 7 Department of Gastrointestinal Oncology, Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education/Beijing), Peking University Cancer Hospital and Institute, Beijing, 100142, China.
  • 8 National Institute of Biological Sciences, Tsinghua Institute of Multidisciplinary Biomedical Research, Tsinghua University, Beijing, 102206, China. [email protected].
  • 9 Department of Immunology, School of Basic Medical Sciences, Beijing Key Laboratory of Tumor Systems Biology, Institute of Systems Biomedicine, Peking University Health Science Center, Beijing, 100191, China. [email protected].
  • # Contributed equally.
PMID: 36726140 DOI: 10.1186/s13059-023-02860-w
Abstract

Background: 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.

Results: Here, we find that knocking down gene expression using RfxCas13d in the adult brain neurons caused death of mice, which may result from the collateral activity of RfxCas13d rather than the loss of target gene function or off-target effects. Mechanistically, we show that RfxCas13d exhibits 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 pathway.

Conclusions: These findings provide new mechanistic insights into the collateral activity of RfxCas13d in mammalian cells and warn that the biosafety of the CRISPR-Cas13 system needs further evaluation before application to clinical treatments.

Keywords

28s rRNA cleavage; Collateral activity; Death of mice; RfxCas13d.

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