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-Structure paper
タイトル | Mechanisms for target recognition and cleavage by the Cas12i RNA-guided endonuclease. |
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ジャーナル・号・ページ | Nat Struct Mol Biol, Vol. 27, Issue 11, Page 1069-1076, Year 2020 |
掲載日 | 2020年9月7日 |
著者 | Heng Zhang / Zhuang Li / Renjian Xiao / Leifu Chang / |
PubMed 要旨 | 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 ...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. |
リンク | Nat Struct Mol Biol / PubMed:32895556 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 2.9 - 3.9 Å |
構造データ | EMDB-21541, PDB-6w5c: EMDB-21551, PDB-6w62: EMDB-21552, PDB-6w64: |
由来 |
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キーワード | HYDROLASE/DNA/RNA / CRISPR / HYDROLASE-DNA-RNA complex |