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PDBsum entry 4oge
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References listed in PDB file
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Key reference
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Title
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Structures of cas9 endonucleases reveal RNA-Mediated conformational activation.
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Authors
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M.Jinek,
F.Jiang,
D.W.Taylor,
S.H.Sternberg,
E.Kaya,
E.Ma,
C.Anders,
M.Hauer,
K.Zhou,
S.Lin,
M.Kaplan,
A.T.Iavarone,
E.Charpentier,
E.Nogales,
J.A.Doudna.
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Ref.
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Science, 2014,
343,
1247997.
[DOI no: ]
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PubMed id
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Abstract
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Type II CRISPR (clustered regularly interspaced short palindromic repeats)-Cas
(CRISPR-associated) systems use an RNA-guided DNA endonuclease, Cas9, to
generate double-strand breaks in invasive DNA during an adaptive bacterial
immune response. Cas9 has been harnessed as a powerful tool for genome editing
and gene regulation in many eukaryotic organisms. We report 2.6 and 2.2 angstrom
resolution crystal structures of two major Cas9 enzyme subtypes, revealing the
structural core shared by all Cas9 family members. The architectures of Cas9
enzymes define nucleic acid binding clefts, and single-particle electron
microscopy reconstructions show that the two structural lobes harboring these
clefts undergo guide RNA-induced reorientation to form a central channel where
DNA substrates are bound. The observation that extensive structural
rearrangements occur before target DNA duplex binding implicates guide RNA
loading as a key step in Cas9 activation.
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