5WQE

Crystal structure of Alicyclobacillus acidoterrestris C2c1 in complex with single-guide RNA at 3.1 Angstrom resolution

Summary for 5WQE

• Entry DOI: 10.2210/pdb5wqe/pdb
• Descriptor: CRISPR-associated endonuclease C2c1, RNA (60-MER) (3 entities in total)
• Functional Keywords: crispr-cas endonuclease, recognition lobe, nuclease lobe, rna binding protein
• Biological source: Alicyclobacillus acidoterrestris; More
• Total number of polymer chains: 2
• Total formula weight: 169244.82

Authors

Liu, L.,Wang, Y.L. (deposition date: 2016-11-26, release date: 2017-01-25, Last modification date: 2024-10-16)

Primary citation

Liu, L.,Chen, P.,Wang, M.,Li, X.,Wang, J.,Yin, M.,Wang, Y.
C2c1-sgRNA Complex Structure Reveals RNA-Guided DNA Cleavage Mechanism
Mol. Cell, 65:310-322, 2017

PubMed Abstract: C2c1 is a type V-B CRISPR-Cas system dual-RNA-guided DNA endonuclease. Here, we report the crystal structure of Alicyclobacillus acidoterrestris C2c1 in complex with a chimeric single-molecule guide RNA (sgRNA). AacC2c1 exhibits a bi-lobed architecture consisting of a REC and NUC lobe. The sgRNA scaffold forms a tetra-helical structure, distinct from previous predictions. The crRNA is located in the central channel of C2c1, and the tracrRNA resides in an external surface groove. Although AacC2c1 lacks a PAM-interacting domain, our analysis revealed that the PAM duplex has a similar binding position found in Cpf1. Importantly, C2c1-sgRNA system is highly sensitive to single-nucleotide mismatches between guide RNA and target DNA. The resulting reduction in off-target cleavage renders C2c1 a valuable addition to the current arsenal of genome-editing tools. Together, our findings indicate that sgRNA assembly is achieved through a mechanism distinct from that reported previously for Cas9 or Cpf1 endonucleases.

PubMed: 27989439
DOI: 10.1016/j.molcel.2016.11.040

Experimental method

X-RAY DIFFRACTION (3.126 Å)