| Autor: |
Sun, Ao, Li, Cheng-Ping, Chen, Zhihang, Zhang, Shouyue, Li, Dan-Yuan, Yang, Yun, Li, Long-Qi, Zhao, Yuqian, Wang, Kaichen, Li, Zhaofu, Liu, Jinxia, Liu, Sitong, Wang, Jia, Liu, Jun-Jie Gogo |
| Zdroj: |
Cell Research; March 2023, Vol. 33 Issue: 3 p229-244, 16p |
| Abstrakt: |
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. |
| Databáze: |
Supplemental Index |
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