Mechanisms for target recognition and cleavage by the Cas12i RNA-guided endonuclease.
Journal
Nature structural & molecular biology
ISSN: 1545-9985
Titre abrégé: Nat Struct Mol Biol
Pays: United States
ID NLM: 101186374
Informations de publication
Date de publication:
11 2020
11 2020
Historique:
received:
09
03
2020
accepted:
06
08
2020
pubmed:
9
9
2020
medline:
29
1
2021
entrez:
8
9
2020
Statut:
ppublish
Résumé
Cas12i is a recently identified type V CRISPR-Cas endonuclease that predominantly cleaves the non-target strand of a double-stranded DNA substrate. This nicking activity of Cas12i could potentially be used for genome editing with high specificity. To elucidate its mechanisms for target recognition and cleavage, we determined cryo-EM structures of Cas12i in multiple functional states. Cas12i pre-orders a seven-nucleotide seed sequence of the crRNA for target recognition and undergoes a two-step activation through crRNA-DNA hybridization. Formation of 14 base pairs activates the nickase activity, and 28-bp hybridization promotes cleavage of the target strand. The atomic structures and mechanistic insights gained should facilitate the manipulation of Cas12i for genome editing applications.
Identifiants
pubmed: 32895556
doi: 10.1038/s41594-020-0499-0
pii: 10.1038/s41594-020-0499-0
pmc: PMC8256696
mid: NIHMS1714499
doi:
Substances chimiques
CRISPR-Associated Proteins
0
RNA
63231-63-0
DNA
9007-49-2
Endonucleases
EC 3.1.-
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1069-1076Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM138675
Pays : United States
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