The compact Casπ (Cas12l) ‘bracelet’ provides a unique structural platform for DNA manipulation
Open accessPreprint — published as 10.1038/s41422-022-00771-2
Abstract
CRISPR-Cas modules serve as the adaptive nucleic-acid immune systems for prokaryotes, and provide versatile tools for nucleic-acid manipulation in various organisms. Here, we discovered a new miniature type V system, CRISPR-Casπ (Cas12l) (∼860 aa), from the environmental metagenome. Complexed with a large guide-RNA (∼170 nt) comprising the tracrRNA and crRNA, Casπ (Cas12l) recognizes a unique 5’ C-rich PAM for DNA cleavage under a broad range of biochemical conditions, and generates gene editing in mammalian cells. Cryo-EM study reveals a ‘bracelet’ architecture of Casπ effector encircling the DNA target at 3.4-Å resolution, substantially different from the canonical ‘two-lobe’ architectures of Cas12 and Cas9 nucleases. And the large guide-RNA serves as a ‘two-arm’ scaffold for effector assembly. Our study expands the knowledge of DNA targeting mechanisms by CRISPR effectors, and offers an efficient but compact platform for DNA manipulations.
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Cites
Highly accurate protein structure prediction with AlphaFold
Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., Tunyasuvunakool, K., Bates, R., Žídek, A., Potapenko, A., et al. (2021) · Nature · doi:10.1038/s41586-021-03819-2 · Cited by 43,764 · see citing works
Protocells and RNA Self-Replication
Joyce, G. F., & Szostak, J. W. (2018) · Cold Spring Harbor Perspectives in Biology · doi:10.1101/cshperspect.a034801 · Cited by 308 · see citing works
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- Authors
- Sun, A., Li, C.-P., Chen, Z., Zhang, S., Li, D., Yang, Y., Li, L.-Q., Zhao, Y., Wang, K., Li, Z., Liu, J., Liu, S., Wang, J., & Liu, J.-J. G.
- Affiliations
- Center for Life Sciences (CN)
- Ministry of Education of the People's Republic of China (CN)
- Corresponding author
- Liu, Jinxia
- Wang, Jia
- ORCID
- Venue
- Cell Research
- Publisher
- openRxiv
- Published
- 2022-12-16
- Volume
- 33
- Issue
- 3
- Pages
- 229-244
- ISSN
- 1001-0602 (ISSN-L)
- PMC
- PMC9977741
- OpenAlex
- W4316810847
- Cited by
- 64
- References
- 2
- Citations by year
- 2026: 15 · 2025: 15 · 2024: 20 · 2023: 14
- FWCI
- 5.68
- Type
- journal article
- Language
- en
- Open access
- green
- doi.org
- Locations
- Cell Research (publishedVersion)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Natural Science Foundation of China (32101195, 32150018)
- Ministry of Science and Technology of the People's Republic of China (2022YFF1002801)
- Tsinghua University
- Ministry of Agriculture and Rural Affairs of the People's Republic of China
- National Key Research and Development Program of China (2022YFF1002801)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- guide RNA, tracrRNA, crRNA, PAM recognition, DNA cleavage, genome editing, metagenome, cryo-EM structure, Cas12, Cas9
- MeSH terms
- Gene Editing, RNA, Guide, CRISPR-Cas Systems, DNA, Endonucleases, CRISPR-Cas Systems
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