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Programmable RNA editing with compact CRISPR-Cas13 systems from uncultivated microbes.
Xu, Chunlong; Zhou, Yingsi; Xiao, Qingquan; He, Bingbing; Geng, Guannan; Wang, Zikang; Cao, Birong; Dong, Xue; Bai, Weiya; Wang, Yifan; Wang, Xiang; Zhou, Dongming; Yuan, Tanglong; Huo, Xiaona; Lai, Jinsheng; Yang, Hui.
  • Xu C; Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Ch
  • Zhou Y; Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Ch
  • Xiao Q; Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Ch
  • He B; College of Life Sciences, University of Chinese Academy of Sciences, Beijing, China.
  • Geng G; Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Ch
  • Wang Z; College of Life Sciences, University of Chinese Academy of Sciences, Beijing, China.
  • Cao B; Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Ch
  • Dong X; Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Ch
  • Bai W; College of Life Sciences, University of Chinese Academy of Sciences, Beijing, China.
  • Wang Y; CAS Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.
  • Wang X; Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Ch
  • Zhou D; Huigene Therapeutics Inc., Shanghai, China.
  • Yuan T; Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Ch
  • Huo X; Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
  • Lai J; Shanghai Public Health Clinical Center, Fudan University, Shanghai, China.
  • Yang H; Department of Pathogen Biology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin, China.
Nat Methods ; 18(5): 499-506, 2021 05.
Article in English | MEDLINE | ID: covidwho-1220210
ABSTRACT
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.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Bacterial / RNA Editing / CRISPR-Cas Systems Limits: Animals / Humans Language: English Journal: Nat Methods Journal subject: Laboratory Techniques and procedures Year: 2021 Document Type: Article Affiliation country: S41592-021-01124-4

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Bacterial / RNA Editing / CRISPR-Cas Systems Limits: Animals / Humans Language: English Journal: Nat Methods Journal subject: Laboratory Techniques and procedures Year: 2021 Document Type: Article Affiliation country: S41592-021-01124-4