@article{d1147bd8dafc4ce6b36ddd3c5addf120,
title = "DNA capture by a CRISPR-Cas9-guided adenine base editor",
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.",
author = "Audrone Lapinaite and Knott, {Gavin J.} and Palumbo, {Cody M.} and Enrique Lin-Shiao and Richter, {Michelle F.} and Zhao, {Kevin T.} and Beal, {Peter A.} and Liu, {David R.} and Doudna, {Jennifer A.}",
note = "Funding Information: Research reported in this publication was supported by the Centers for Excellence in Genomic Science of the National Institutes of Health under award no. RM1HG009490 and the Somatic Cell Genome Editing Program of the Office of the Director, National Institutes of Health (OD), under award no. U01AI142817-02. J.A.D. receives funding from the William M. Keck Foundation, the National Multiple Sclerosis Society, the Paul Allen Frontiers Group, and the Howard Hughes Medical Institute. This material is based upon work supported by the National Science Foundation under award no. 1817593. A.L. was supported by the Centers for Excellence in Genomic Science of the National Institutes of Health under award no. RM1HG009490. G.J.K. was supported by an NHMRC Investigator Grant (EL1, 1175568) and previously by an American Australian Association Fellowship. E.L.-S. was supported by the Paul G. Allen Frontiers Group. P.A.B. was supported by NIH grant no. R01GM061115. C.M.P. was supported by NIH Predoctoral Training Grant no. T32GM113770. D.R.L. was supported by NIH grant nos. U01 AI142756, RM1 HG009490, R01 EB022376, and R35 GM118062; the St. Jude Collaborative Research Consortium; the Bill and Melinda Gates Foundation; and the Howard Hughes Medical Institute. M.F.R. was supported by an HHMI Hanna Gray Fellowship. K.T.Z. was supported by Harvard Chemical Biology Training Grant no. T32 GM095450. Publisher Copyright: {\textcopyright} 2020 American Association for the Advancement of Science. All rights reserved.",
year = "2020",
month = jul,
day = "31",
doi = "10.1126/science.abb1390",
language = "English (US)",
volume = "369",
pages = "566--571",
journal = "Science",
issn = "0036-8075",
publisher = "American Association for the Advancement of Science",
number = "6503",
}