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The Molecular Architecture for RNA-Guided RNA Cleavage by Cas13a
Abstract
Cas13a, a type VI-A CRISPR-Cas RNA-guided RNA ribonuclease, degrades invasive RNAs targeted by CRISPR RNA (crRNA) and has potential applications in RNA technology. To understand how Cas13a is activated to cleave RNA, we have determined the crystal structure of Leptotrichia buccalis (Lbu) Cas13a bound to crRNA and its target RNA, as well as the cryo-EM structure of the LbuCas13a-crRNA complex. The crRNA-target RNA duplex binds in a positively charged central channel of the nuclease (NUC) lobe, and Cas13a protein and crRNA undergo a significant conformational change upon target RNA binding. The guide-target RNA duplex formation triggers HEPN1 domain to move toward HEPN2 domain, activating the HEPN catalytic site of Cas13a protein, which subsequently cleaves both single-stranded target and collateral RNAs in a non-specific manner. These findings reveal how Cas13a of type VI CRISPR-Cas systems defend against RNA phages and set the stage for its development as a tool for RNA manipulation.
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In the citing paper
“Upon linking to the target, Cas13 RNase activity is prompted by the approximation of the catalytic sites of both HEPN domains [59].”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [59]
In the citing paper
“Cas9 is guided by a 20 nt fraction of the gRNA”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [59]
In the citing paper
“and it maintains cleaving activity even with 3-5 mismatches at the PAM-distal end of the gRNA molecule [59,81,102].”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [59]
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Metadata
- Authors
- Liu, L., Li, X., Ma, J., Li, Z., You, L., Wang, J., Wang, M., Zhang, X., & Wang, Y.
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2017-08
- Volume
- 170
- Issue
- 4
- Pages
- 714-726.e10
- ISSN
- 0092-8674
- Cited by
- 538
- References
- 3
- Language
- en
- Licence
- Funders
- Salt Science Research Foundation
- National Natural Science Foundation of China (31400640, 31570874, 31571335, 91440201)
- Chinese Academy of Sciences (XDB08010203)
- Ministry of Science and Technology of the People's Republic of China (2014CB910102)
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