6VPC

Structure of the SpCas9 DNA adenine base editor - ABE8e

6VPC の概要

• エントリーDOI: 10.2210/pdb6vpc/pdb
• EMDBエントリー: 21308
• 分子名称: Cas9 (SpCas9) single-guide RNA (sgRNA), CRISPR-associated endonuclease Cas9, DNA target strand (TS), ... (7 entities in total)
• 機能のキーワード: base editor, abe, cas9, crispr, dna binding protein-dna-rna complex, dna binding protein/dna/rna
• 由来する生物種: Streptococcus pyogenes; 詳細
• タンパク質・核酸の鎖数: 6
• 化学式量合計: 264575.83

構造登録者

Knott, G.J.,Lapinaite, A.,Doudna, J.A. (登録日: 2020-02-03, 公開日: 2020-07-29, 最終更新日: 2024-03-06)

主引用文献

Lapinaite, A.,Knott, G.J.,Palumbo, C.M.,Lin-Shiao, E.,Richter, M.F.,Zhao, K.T.,Beal, P.A.,Liu, D.R.,Doudna, J.A.
DNA capture by a CRISPR-Cas9-guided adenine base editor.
Science, 369:566-571, 2020

PubMed Abstract: CRISPR-Cas-guided base editors convert A•T to G•C, or C•G to T•A, in cellular DNA for precision genome editing. To understand the molecular basis for DNA adenosine deamination by adenine base editors (ABEs), we determined a 3.2-angstrom resolution cryo-electron microscopy structure of ABE8e in a substrate-bound state in which the deaminase domain engages DNA exposed within the CRISPR-Cas9 R-loop complex. Kinetic and structural data suggest that ABE8e catalyzes DNA deamination up to ~1100-fold faster than earlier ABEs because of mutations that stabilize DNA substrates in a constrained, transfer RNA-like conformation. Furthermore, ABE8e's accelerated DNA deamination suggests a previously unobserved transient DNA melting that may occur during double-stranded DNA surveillance by CRISPR-Cas9. These results explain ABE8e-mediated base-editing outcomes and inform the future design of base editors.

PubMed: 32732424
DOI: 10.1126/science.abb1390

実験手法

ELECTRON MICROSCOPY (3.2 Å)