Journal
NATURE STRUCTURAL & MOLECULAR BIOLOGY
Volume 27, Issue 11, Pages 1069-+Publisher
NATURE RESEARCH
DOI: 10.1038/s41594-020-0499-0
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Funding
- NIH [R01GM138675]
- Showalter Trust Research Award
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Cryo-EM structures of Cas12i in multiple functional states provide insights that might facilitate the manipulation of this type V CRISPR-Cas endonuclease for genome editing applications. 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.
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