+Search query
-Structure paper
Title | The compact Casπ (Cas12l) 'bracelet' provides a unique structural platform for DNA manipulation. |
---|---|
Journal, issue, pages | Cell Res, Vol. 33, Issue 3, Page 229-244, Year 2023 |
Publish date | Jan 17, 2023 |
Authors | Ao 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 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 ...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 links | Cell Res / PubMed:36650285 / PubMed Central |
Methods | EM (single particle) |
Resolution | 3.36 Å |
Structure data | EMDB-33983, PDB-7yoj: |
Source |
|
Keywords | RNA BINDING PROTEIN/DNA/RNA / CasPi complex / RNA BINDING PROTEIN / RNA BINDING PROTEIN-DNA-RNA complex |