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-Structure paper
タイトル | Exploiting activation and inactivation mechanisms in type I-C CRISPR-Cas3 for genome-editing applications. |
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ジャーナル・号・ページ | Mol Cell, Vol. 84, Issue 3, Page 463-475.e5, Year 2024 |
掲載日 | 2024年2月1日 |
著者 | Chunyi Hu / Mason T Myers / Xufei Zhou / Zhonggang Hou / Macy L Lozen / Ki Hyun Nam / Yan Zhang / Ailong Ke / |
PubMed 要旨 | Type I CRISPR-Cas systems utilize the RNA-guided Cascade complex to identify matching DNA targets and the nuclease-helicase Cas3 to degrade them. Among the seven subtypes, type I-C is compact in size ...Type I CRISPR-Cas systems utilize the RNA-guided Cascade complex to identify matching DNA targets and the nuclease-helicase Cas3 to degrade them. Among the seven subtypes, type I-C is compact in size and highly active in creating large-sized genome deletions in human cells. Here, we use four cryoelectron microscopy snapshots to define its RNA-guided DNA binding and cleavage mechanisms in high resolution. The non-target DNA strand (NTS) is accommodated by I-C Cascade in a continuous binding groove along the juxtaposed Cas11 subunits. Binding of Cas3 further traps a flexible bulge in NTS, enabling NTS nicking. We identified two anti-CRISPR proteins AcrIC8 and AcrIC9 that strongly inhibit Neisseria lactamica I-C function. Structural analysis showed that AcrIC8 inhibits PAM recognition through allosteric inhibition, whereas AcrIC9 achieves so through direct competition. Both Acrs potently inhibit I-C-mediated genome editing and transcriptional modulation in human cells, providing the first off-switches for type I CRISPR eukaryotic genome engineering. |
リンク | Mol Cell / PubMed:38242128 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.0 - 3.64 Å |
構造データ | EMDB-29877, PDB-8g9s: EMDB-29878, PDB-8g9t: EMDB-29879, PDB-8g9u: EMDB-29896, PDB-8gaf: EMDB-29900, PDB-8gam: EMDB-29901, PDB-8gan: |
化合物 | ChemComp-MG: ChemComp-PO4: |
由来 |
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キーワード | HYDROLASE/RNA / CRISPR / type I-C / cascade / anti-CRISPR / HYDROLASE-RNA complex / DNA BINDING PROTEIN / HYDROLASE/RNA/DNA / HYDROLASE-RNA-DNA complex |