文献类型： 外文期刊
作者： Xu, Chunlong 1 ; Zhou, Yingsi 1 ; Xiao, Qingquan 1 ; He, Bingbing 1 ; Geng, Guannan 1 ; Wang, Zikang 1 ; Cao, Birong 2 ;
作者机构： 1.Chinese Acad Sci, Shanghai Res Ctr Brain Sci & Brain Inspired Intel, Shanghai Inst Biol Sci,CAS Ctr Excellence Brain S, Inst Neurosci,State Key Lab Neurosci,Key Lab Prim, Shanghai, Peoples R China
2.Univ Chinese Acad Sci, Coll Life Sci, Beijing, Peoples R China
3.Chinese Acad Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, Shanghai, Peoples R China
4.Huigene Therapeut Inc, Shanghai, Peoples R China
5.Fudan Univ, Shanghai Publ Hlth Clin Ctr, Shanghai, Peoples R China
6.Tianjin Med Univ, Sch Basic Med Sci, Dept Pathogen Biol, Tianjin, Peoples R China
7.Chinese Acad Agr Sci, Agr Genome Inst Shenzhen, Ctr Anim Genom, Shenzhen, Peoples R China
8.China Agr Univ, Dept Plant Genet & Breeding, State Key Lab Agrobiotechnol, Beijing, Peoples R China
9.China Agr Univ, Dept Plant Genet & Breeding, Natl Maize Improvement Ctr, Beijing, Peoples R China
期刊名称：NATURE METHODS （ 影响因子：30.822； 五年影响因子：36.147 ）
ISSN： 1548-7091
年卷期：
页码：
收录情况： SCI
摘要： Competitive coevolution between microbes and viruses has led to the diversification of CRISPR-Cas defense systems against infectious agents. By analyzing metagenomic terabase datasets, we identified two compact families (775 to 803 amino acids (aa)) of CRISPR-Cas ribonucleases from hypersaline samples, named Cas13X and Cas13Y. We engineered Cas13X.1 (775 aa) for RNA interference experiments in mammalian cell lines. We found Cas13X.1 could tolerate single-nucleotide mismatches in RNA recognition, facilitating prophylactic RNA virus inhibition. Moreover, a minimal RNA base editor, composed of engineered deaminase (385 aa) and truncated Cas13X.1 (445 aa), exhibited robust editing efficiency and high specificity to induce RNA base conversions. Our results suggest that there exist untapped bacterial defense systems in natural microbes that can function efficiently in mammalian cells, and thus potentially are useful for RNA-editing-based research. This work describes the identification of Cas13 proteins from two families by mining public metagenomic data. The newly identified Cas13X.1 shows efficient target knockdown and can be used to degrade SARS-CoV-2 and H1N1 genomes. In addition, the truncated Cas13X.1 offers an advantage in generating mini-RNA base editors.
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