6VPC
Structure of the SpCas9 DNA adenine base editor - ABE8e
Summary for 6VPC
| Entry DOI | 10.2210/pdb6vpc/pdb |
| EMDB information | 21308 |
| Descriptor | Cas9 (SpCas9) single-guide RNA (sgRNA), CRISPR-associated endonuclease Cas9, DNA target strand (TS), ... (7 entities in total) |
| Functional Keywords | base editor, abe, cas9, crispr, dna binding protein-dna-rna complex, dna binding protein/dna/rna |
| Biological source | Streptococcus pyogenes More |
| Total number of polymer chains | 6 |
| Total formula weight | 264575.83 |
| Authors | Knott, G.J.,Lapinaite, A.,Doudna, J.A. (deposition date: 2020-02-03, release date: 2020-07-29, Last modification date: 2024-03-06) |
| Primary citation | 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 Cited by 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: 32732424DOI: 10.1126/science.abb1390 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3.2 Å) |
Structure validation
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