EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Improving the efficiency of high-fidelity Cas9 by enhancing PAM-distal interactions.
ジャーナル・号・ページ	Nat Struct Mol Biol, Vol. 33, Issue 4, Page 590-602, Year 2026
掲載日	2026年3月18日
著者	Rong Zheng / Zhike Lu / Rongwei Wei / Young-Cheul Shin / Jiang Du / Qingfeng Zhang / Jianbo Li / Xiaoqi Wang / Yi Wei / Botao Liu / Yang Chen / Lihong Ding / Heng Zhang / Hui Chen / Jing Huang / Lijia Ma /
PubMed 要旨	Engineering CRISPR enzymes for high fidelity often impairs cleavage activity. Meanwhile, a mechanistic understanding of why high-fidelity mutations reduce Cas9's cleavage activity remains unclear, ...Engineering CRISPR enzymes for high fidelity often impairs cleavage activity. Meanwhile, a mechanistic understanding of why high-fidelity mutations reduce Cas9's cleavage activity remains unclear, presenting a challenge in balancing nuclease specificity and efficiency for clinical applications. In this study, we show that extending the spacer region to 21 or 22 nucleotides restores the impaired cleavage activity of SuperFi-Cas9, a high-fidelity Cas9 variant with 7 mutations in the RuvC domain at the protospacer adjacent motif (PAM)-distal region. Cryo-electron microscopy structures and mutational analyses reveal that the negatively charged mutations in a protruding loop of the RuvC domain create repulsive forces that destabilize the nuclease-single guide (sg)RNA-DNA complex. Spacer extension enhances interactions in the PAM-distal region, effectively restoring cleavage activity and balancing editing efficiency with specificity. In addition, we develop a deep learning model, AIdit-SuperFi, to predict optimal sgRNA length for high-fidelity genome editing. Our findings introduce a straightforward strategy to enhance CRISPR complex stability and provide mechanistic insights into the impaired cleavage activity of engineered high-fidelity Cas9, presenting a pathway toward precise and efficient genome editing and clinical translation of CRISPR technologies.
リンク	Nat Struct Mol Biol / PubMed:41851507
手法	EM (単粒子)
解像度	3.27 - 4.4 Å
構造データ	62054 9k4c EMDB-62054, PDB-9k4c: SuperFi Cas9 - 22nt sgRNA - DNA ternary complex 手法: EM (単粒子) / 解像度: 3.84 Å 62055 9k4d EMDB-62055, PDB-9k4d: SuperFi Cas9 - 20nt sgRNA - DNA ternary complex 手法: EM (単粒子) / 解像度: 3.53 Å 62056 9k4e EMDB-62056, PDB-9k4e: SuperFi Cas9 - mismatch 22nt sgRNA - DNA ternary complex class1 手法: EM (単粒子) / 解像度: 3.3 Å 62057 9k4f EMDB-62057, PDB-9k4f: SuperFi Cas9 - mismatch 22nt sgRNA - DNA ternary complex class2 手法: EM (単粒子) / 解像度: 3.27 Å 62059 9k4h EMDB-62059, PDB-9k4h: SuperFi Cas9 - mismatch 22nt sgRNA - DNA ternary complex class3 手法: EM (単粒子) / 解像度: 3.36 Å 65730 9w7q EMDB-65730, PDB-9w7q: SuperFi Cas9 - 20nt sgRNA - DNA ternary complex Class A 手法: EM (単粒子) / 解像度: 3.79 Å 65732 9w7t EMDB-65732, PDB-9w7t: SuperFi Cas9 - 20nt sgRNA - DNA ternary complex Class B 手法: EM (単粒子) / 解像度: 3.91 Å 65733 9w7u EMDB-65733, PDB-9w7u: SuperFi Cas9 - 20nt sgRNA - DNA ternary complex Class C 手法: EM (単粒子) / 解像度: 3.83 Å 65734 9w7v EMDB-65734, PDB-9w7v: SuperFi Cas9 - 20nt sgRNA - DNA ternary complex Class D 手法: EM (単粒子) / 解像度: 3.81 Å 65771 9w9d EMDB-65771, PDB-9w9d: SuperFi Cas9 - 22nt sgRNA - DNA ternary complex 手法: EM (単粒子) / 解像度: 4.4 Å 65809 9wa9 EMDB-65809, PDB-9wa9: SuperFi Cas9 - 22nt sgRNA - DNA ternary complex Class B 手法: EM (単粒子) / 解像度: 4.12 Å 65810 9waa EMDB-65810, PDB-9waa: SuperFi Cas9 - 22nt sgRNA - DNA ternary complex Class C 手法: EM (単粒子) / 解像度: 4.17 Å 65827 9waw EMDB-65827, PDB-9waw: SuperFi Cas9 - 20nt sgRNA - DNA ternary complex Class E 手法: EM (単粒子) / 解像度: 3.95 Å
由来	streptococcus pyogenes (化膿レンサ球菌) synthetic construct (人工物) streptococcus pyogenes serotype m1 (化膿レンサ球菌)
キーワード	IMMUNE SYSTEM/DNA/RNA / Complex / cas / SuperFi cas9 / IMMUNE SYSTEM / IMMUNE SYSTEM-DNA-RNA complex