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TitleThe compact Casπ (Cas12l) 'bracelet' provides a unique structural platform for DNA manipulation.
Journal, issue, pagesCell Res, Vol. 33, Issue 3, Page 229-244, Year 2023
Publish dateJan 17, 2023
AuthorsAo Sun / Cheng-Ping Li / Zhihang Chen / Shouyue Zhang / Dan-Yuan Li / Yun Yang / Long-Qi Li / Yuqian Zhao / Kaichen Wang / Zhaofu Li / Jinxia Liu / Sitong Liu / Jia Wang / Jun-Jie Gogo Liu /
PubMed AbstractCRISPR-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 ...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. 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 manipulation.
External linksCell Res / PubMed:36650285 / PubMed Central
MethodsEM (single particle)
Resolution3.36 Å
Structure data

33983

7yoj

EMDB-33983, PDB-7yoj:
Structure of CasPi with guide RNA and target DNA
Method: EM (single particle) / Resolution: 3.36 Å

Source
  • armatimonadetes bacterium (bacteria)
  • armatimonadota bacterium (bacteria)
KeywordsRNA BINDING PROTEIN/DNA/RNA / CasPi complex / RNA BINDING PROTEIN / RNA BINDING PROTEIN-DNA-RNA complex

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